Details of a Researcher - FURUHASHI Masato

Papers - FURUHASHI Masato

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Machine learning-based analyses of contributing factors for the development of hypertension: a comparative study.

Marenao Tanaka, Yukinori Akiyama, Kazuma Mori, Itaru Hosaka, Keisuke Endo, Toshifumi Ogawa, Tatsuya Sato, Toru Suzuki, Toshiyuki Yano, Hirofumi Ohnishi, Nagisa Hanawa, Masato Furuhashi

Clinical and experimental hypertension (New York, N.Y. : 1993) 47 ( 1 ) 2449613 - 2449613 2025.12 [International journal]

OBJECTIVES: Sufficient attention has not been given to machine learning (ML) models using longitudinal data for investigating important predictors of new onset of hypertension. We investigated the predictive ability of several ML models for the development of hypertension. METHODS: A total of 15 965 Japanese participants (men/women: 9,466/6,499, mean age: 45 years) who received annual health examinations were randomly divided into a training group (70%, n = 11,175) and a test group (30%, n = 4,790). The predictive abilities of 58 candidates including fatty liver index (FLI), which is calculated by using body mass index, waist circumference and levels of γ-glutamyl transferase and triglycerides, were investigated by statistics analogous to the area under the curve (AUC) in receiver operating characteristic curve analyses using ML models including logistic regression, random forest, naïve Bayes, extreme gradient boosting and artificial neural network. RESULTS: During a 10-year period (mean period: 6.1 years), 2,132 subjects (19.1%) in the training group and 917 subjects (19.1%) in the test group had new onset of hypertension. Among the 58 parameters, systolic blood pressure, age and FLI were identified as important candidates by random forest feature selection with 10-fold cross-validation. The AUCs of ML models were 0.765-0.825, and discriminatory capacity was significantly improved in the artificial neural network model compared to that in the logistic regression model. CONCLUSIONS: The development of hypertension can be simply and accurately predicted by each ML model using systolic blood pressure, age and FLI as selected features. By building multiple ML models, more practical prediction might be possible.

Can small dense LDL cholesterol be estimated from the lipid profile?

Tatsuya Sato, Marenao Tanaka, Masato Furuhashi

Current opinion in lipidology 2025.04 [International journal]

PURPOSE OF REVIEW: Small dense low-density lipoprotein cholesterol (sdLDL-C) is recognized for its strong atherosclerogenic potential. However, its direct measurement remains impractical in clinical settings due to its high cost, time constraints, and labor-intensive nature. This review discusses the benefits and limitations of estimating sdLDL-C using conventional lipid fractions, highlighting recent advancements in estimation methods. RECENT FINDINGS: Sampson et al. proposed a novel equation for estimating sdLDL-C based on conventional lipid parameters, offering a more accessible alternative to direct measurement. Recent studies, including ours, demonstrated that this estimation method achieves sufficiently high accuracy for overall application. However, its accuracy can be improved by incorporating machine learning. Furthermore, sdLDL-C estimated by Sampson's equation has been shown to be a superior risk marker for hypertension, an intermediate phenotype of atherosclerosis, and ischemic heart disease, a major cardiovascular event, compared to conventional lipid profiles alone, although further research is needed to determine whether estimated sdLDL-C is equivalent to directly measured sdLDL-C in risk assessment. SUMMARY: Estimated sdLDL-C presents a promising alternative to direct measurement. While estimated sdLDL-C levels can serve a risk marker for cardiovascular diseases, further research is needed to refine estimation models and explore their integration into clinical practice.

Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) Is an Independent Risk Factor for the Development of Ischemic Heart Disease ― A 10-Year Cohort Study ―

Toshifumi Ogawa, Tatsuya Sato, Marenao Tanaka, Yukinori Akiyama, Kei Nakata, Hidemichi Kouzu, Kazuma Mori, Hiroki Aida, Wataru Kawaharata, Itaru Hosaka, Toru Suzuki, Nagisa Hanawa, Masato Furuhashi

Circulation Reports ( Japanese Circulation Society ) 2025.04

High Level of Estimated Small Dense Low-Density Lipoprotein Cholesterol as an Independent Risk Factor for the Development of Ischemic Heart Disease Regardless of Low-Density Lipoprotein Cholesterol Level ― A 10-Year Cohort Study ―

Keisuke Endo, Marenao Tanaka, Tatsuya Sato, Masafumi Inyaku, Kei Nakata, Wataru Kawaharata, Hiroki Aida, Itaru Hosaka, Yukinori Akiyama, Nagisa Hanawa, Masato Furuhashi

Circulation Journal ( Japanese Circulation Society ) 2025.03

Elevated urinary fatty acid-binding protein 4 level predicts future renal dysfunction and poor prognosis in Japanese patients with diabetes: a longitudinal cohort study

Marenao Tanaka, Tatsuya Sato, Tomohito Gohda, Nozomu Kamei, Maki Murakoshi, Erika Ishiwata, Kei Nakata, Yukinori Akiyama, Keisuke Endo, Wataru Kawaharata, Hiroki Aida, Toru Suzuki, Mitsunobu Kubota, Michiyoshi Sanuki, Yusuke Suzuki, Masato Furuhashi

Clinical Kidney Journal ( Oxford University Press (OUP) ) 2025.03

Abstract Background Fatty acid-binding protein 4 (FABP4) is an adipokine secreted from adipocytes and macrophages and is also expressed in injured, but not normal, glomerular endothelial cells. Elevated levels of urinary FABP4 (U-FABP4) have been reported to be associated with glomerular damage and increased proteinuria. Methods The associations of levels of U-FABP4 at baseline with future events including renal dysfunction defined by a 30% decline in estimated glomerular filtration rate (eGFR) and all-cause death were investigated in 660 patients with diabetes (type 1/2: 57/603). Results During a follow-up period (median: 62 months), 90 patients (13.6%) developed renal dysfunction, and 66 patients (10.0%) died (median follow-up period: 65 months). Kaplan-Meier survival curves showed that there were significant differences in cumulative incidences for a 30% decline in eGFR and all-cause death in patients divided by the tertiles of U-FABP4 level. Furthermore, multivariable Cox proportional hazard models with a restricted cubic spline showed that hazard ratios for a 30% decline in eGFR and all-cause death increased with a higher level of logarithmically transformed (Log) U-FABP4 after adjustment for age, sex, type of diabetes, body mass index, current smoking habit, duration of diabetes, comorbidities of hypertension and dyslipidemia, eGFR and the categorical classification of urinary albumin-creatinine ratio. The addition of Log U-FABP4 to traditional risk factors significantly increased the discriminatory capacities for renal dysfunction in net reclassification improvement (NRI) and integrated discrimination improvement and for all-cause death in NRI. Conclusion U-FABP4 is a predictive biomarker for future renal dysfunction and poor prognosis in patients with diabetes.

The Combination of PPARα Agonist GW7647 and Imeglimin Has Potent Effects on High-Glucose-Induced Cellular Biological Responses in Human Retinal Pigment Epithelium Cells.

Nami Nishikiori, Megumi Watanabe, Megumi Higashide, Araya Umetsu, Toshifumi Ogawa, Masato Furuhashi, Hiroshi Ohguro, Tatsuya Sato

Bioengineering (Basel, Switzerland) 12 ( 3 ) 2025.03 [International journal]

BACKGROUND: Hyperglycemic changes in the cellular biological properties of retinal pigment epithelium cells are involved in the pathophysiology of diabetic retinopathy (DR). To assess the effects of the new anti-diabetic agent imeglimin (Ime) on DR, the pharmacological effects of Ime and those of metformin (Met) in combination with the PPARα agonist GW7646 (GW) on adult retinal pigment epithelium (ARPE19) cells cultured in high-glucose conditions were compared. METHODS: Cell viability, levels of reactive oxygen species (ROS), monolayer barrier function measured by transepit very much helial electrical resistance (TEER), and metabolic functions determined by an extracellular flux analyzer were evaluated. RESULTS: While glucose concentrations did not alter cell viability regardless of the presence of Met or Ime, levels of ROS were significantly increased by the high-glucose conditions, and increased levels of ROS were significantly alleviated by the combination of Ime and GW but not by Met alone. Similarly, TEER values were increased by high-glucose conditions, but the effects of high-glucose conditions were dramatically enhanced by the combination of Ime and GW. Furthermore, a metabolic assay showed that an energetic shift was induced by the combination of Ime and GW, whereas energy status became quiescent with Met or Ime alone. CONCLUSIONS: The collective results suggest that Ime in combination with GW has synergetic effects on high-glucose-induced cellular biological changes in ARPE19 cells.

Construction of Predictive Models for Cardiovascular Mortality by Machine Learning Approaches in Patients Who Underwent Transcatheter Aortic Valve Implantation

Shunsaku Otomo, Itaru Hosaka, Marenao Tanaka, Naoto Murakami, Nobuaki Kokubu, Atsuko Muranaka, Ryo Nishikawa, Naoki Hachiro, Ryota Kawamura, Jun Nakata, Nobutaka Nagano, Yukinori Akiyama, Tatsuya Sato, Yutaka Iba, Toshiyuki Yano, Nobuyoshi Kawaharada, Masato Furuhashi

Circulation Reports ( Japanese Circulation Society ) 2025.03

Expression and Secretion of Intraocular Fatty Acid-Binding Protein 4 (ioFABP4) and 5 (ioFABP5) Are Regulated by Glucose Levels and Fatty Acids.

Hiroshi Ohguro, Megumi Higashide, Erika Ishiwata, Fumihito Hikage, Megumi Watanabe, Nami Nishikiori, Tatsuya Sato, Masato Furuhashi

International journal of molecular sciences 26 ( 5 ) 2025.02 [International journal]

Intraocularly, fatty acid-binding protein 4 (FABP4) and 5 (FABP5) mainly originate from human ocular choroidal fibroblasts (HOCF), and human nonpigmented ciliary epithelium (HNPCE) cells have been suggested to be pivotally involved in intraocular pathophysiology. To elucidate the unidentified regulatory mechanisms of the gene expression and protein secretion of FABPs, the effects of glucose levels, fatty acids (FAs), and peroxisome proliferator-activated receptor (PPAR) modulators were studied. To elucidate the additional biological role of FABPs, laser choroidal neovascularization (CNV) in Fabp4-/- and Fabp4/5-/- mice was analyzed by fluorescein angiography. By changing glucose levels, the secretion and expression of FABP4 in HOCF were significantly upregulated, whereas the secretion and expression of FABP5 in HNPCE decreased. The administration of various FAs, particularly docosahexaenoic acid (DHA), markedly increased the expression and secretion of both FABPs. PPAR modulators also influenced the secretion and expression of FABPs. In vivo, wild-type retina exhibited evident CNV with high fluorescein intensity, while Fabp4-/- retina showed reduced CNV formation and Fabp4/5-/- retina displayed evident CNV along with vitreous leakage. These findings suggest that (1) the production and secretion of intraocular FABP4 and FABP5 are distinctly regulated by glucose levels, FAs, and PPARs; and (2) intraocular FABP4 and FABP5 are critical for inducing retinal neovascularization and maintaining the blood-aqueous barrier.

Unraveling Novel Subsets of Lymphocytes Involved in Sac Expansion in the Tertiary Lymphoid Structure Within an Abdominal Aortic Aneurysm.

Itaru Hosaka, Ippei Ikegami, Takuma Mikami, Tatsuya Sato, Toshifumi Ogawa, Kei Mukawa, Marenao Tanaka, Keisuke Endo, Yukinori Akiyama, Akihito Ohkawa, Junji Nakazawa, Tsuyoshi Shibata, Tomohiro Nakajima, Yutaka Iba, Chikara Shiiku, Satoshi Sumino, Ryuji Koshima, Kenichi Takano, Shingo Ichimiya, Nobuyoshi Kawaharada, Masato Furuhashi

Journal of the American Heart Association e040279 2025.02 [International journal]

BACKGROUND: Chronic inflammation is involved in the development of abdominal aortic aneurysm (AAA). A tertiary lymphoid structure (TLS) within vascular lesions has recently been focused on for its role in modulation of inflammation in local tissues. We aimed to elucidate the relationships between TLS and pathophysiology of AAA. METHODS: Abdominal aortic samples obtained from 37 patients with AAA (men/women: 34/3, age: 72.8±9.9 years) and 15 autopsied patients who died from non-aortic events (men/women: 11/4, age: 65.5±9.8 years) were investigated. RESULTS: TLSs in AAA lesions were confirmed by focal infiltration of CD3-positive cells surrounding germinal center-like structures containing CD20-positive cells between the tunica adventitia and tunica media layers. The formation of a TLS was significantly more prevalent in AAA patients than in autopsied patients. The number of TLSs in AAA lesions was positively correlated with sac diameter (r=0.357, P=0.035) and the amount of intraluminal thrombosis (r=0.466, P=0.005). T cells and B cells were predominant cellular populations among CD45+ cells in AAA lesions. There was a significantly positive correlation between the proportions of interfollicular T follicular helper (CD3+CD4+CD45RA-CXCR5+PD-1+) cells and double negative B (CD3-CD19+IgD-CD27-) cells, and they were positively correlated with sac diameter, intraluminal thrombosis, and serum lipids. Deposited single-cell RNA-sequencing data for AAA showed that T follicular helper cells and double negative B cells were associated with lipid metabolism, T cell activation/proliferation and inflammation. CONCLUSIONS: The formation of a TLS in AAA lesions is associated with sac diameter and intraluminal thrombosis in connection with interfollicular T follicular helper cells and double negative B cells, which may contribute to the pathophysiology of AAA and might be novel therapeutic targets for the development of AAA.

High-Glucose-Induced Metabolic and Redox Alterations Are Distinctly Modulated by Various Antidiabetic Agents and Interventions Against FABP5/7, MITF and ANGPTL4 in Melanoma A375 Cells

Nami Nishikiori, Hiroshi Ohguro, Megumi Watanabe, Megumi Higashide, Toshifumi Ogawa, Masato Furuhashi, Tatsuya Sato

International Journal of Molecular Sciences ( MDPI AG ) 26 ( 3 ) 1014 - 1014 2025.01

Hyperglycemia-induced effects on cellular metabolic properties and reactive oxygen species (ROS) generation play pivotal roles in the pathogenesis of malignant melanoma (MM). This study assessed how metabolic states, ROS production, and related gene expression are modulated by antidiabetic agents. The anti-diabetic agents metformin (Met) and imeglimin (Ime), inhibitors of fatty acid-binding proteins 5/7 (MF6) and microphthalmia-associated transcription factor (MITF) (ML329), and siRNA-mediated knockdown of angiopoietin-like protein 4 (ANGPTL4), which affect mitochondrial respiration, ROS production, and related gene expression, were tested in A375 (MM cell line) cells cultured in low (5.5 mM) and high glucose (50 mM) conditions. Cellular metabolic functions were significantly and differently modulated by Met, Ime, MF6, or ML329 and knockdown of ANGPTL4. High glucose significantly enhanced ROS production, which was alleviated by Ime but not by Met. Both MF6 and ML329 reduced ROS levels under both low and high glucose conditions. Knockdown of ANGPTL4 enhanced the change in glucose-dependent ROS production. Gene expression related to mitochondrial respiration and the pathogenesis of MM was significantly modulated by different glucose conditions, antidiabetic agents, MF6, and ML329. These findings suggest that glucose-dependent changes in cellular metabolism and redox status are differently modulated by antidiabetic agents, inhibition of fatty acid-binding proteins or MITF, and ANGPTL4 knockdown in A375 cells.

Glomerular diameter is associated with a reduction in urinary protein by treatment with dapagliflozin in patients with chronic kidney disease

Arata Osanami, Hiroaki Komatsu, Yufu Gocho, Keitaro Nishizawa, Marenao Tanaka, Yuichi Nakamura, Masato Furuhashi

Clinical and Experimental Nephrology ( Springer Science and Business Media LLC ) 2025.01

Effects of linsitinib on M22 and IGF:1-treated 3D spheroids of human orbital fibroblasts.

Fumihito Hikage, Megumi Suzuki, Tatsuya Sato, Araya Umetsu, Toshifumi Ogawa, Nami Nishikiori, Masato Furuhashi, Hiroshi Ohguro, Megumi Watanabe

Scientific reports 15 ( 1 ) 384 - 384 2025.01 [International journal]

To elucidate the role of IGF1R inhibition in the pathogenesis of Graves' orbitopathy (GO), the effects of linsitinib (Lins) on a recombinant human TSHR antibody (M22) and IGF1 to activate TSHR and IGF1R of human orbital fibroblasts (HOFs) obtained from patients without GO (HOFs) and patients with GO (GHOFs) were studied using in vitro three-dimensional (3D) spheroid models in addition to their 2D planar cell culture. For this purpose, we evaluated 1) cellular metabolic functions by using a seahorse bioanalyzer (2D), 2) physical properties including size and stiffness of 3D spheroids, and mRNA expression of several extracellular matrix (ECM) proteins, their modulators (CCL2 LOX, CTGF, MMPs), ACTA2 and inflammatory cytokines (IL1β, IL6). Administration of IGF1 and M22 induced increases of cellular metabolic functions with the effect on HOFs being much more potent than the effect on GHOFs, suggesting that IGF1R and TSHR of GHOFs may already be stimulated. Lins had effects similar to those of IGF1/M22 on cellular biological functions of HOFs but not on those of GHOFs. As for physical properties of 3D GHOFs spheroids, stiffness but not size was significantly increased by IGF1 and/or M22. In contrast, Lins significantly inhibited the M22-induced increase in stiffness despite the fact that Lins alone had no effect. The mRNA expression levels of several genes of ECM proteins and most of the other genes also fluctuated similarly to the changes in stiffness of 3D spheroids despite the fact that Lins induced up-regulation of inflammatory cytokines and MMP3. The findings presented herein indicate that IGF1R inhibition by Lins may beneficially affect GO-related fibrogenesis.

Prognostic implication of sarcopenia diagnosed by updated Asian Working Group for Sarcopenia criteria in older patients with heart failure: Utility and limitation

Satoshi Katano, Kotaro Yamano, Toshiyuki Yano, Ryo Numazawa, Ryohei Nagaoka, Suguru Honma, Yusuke Fujisawa, Yasuhiro Miki, Yuhei Takamura, Hayato Kunihara, Hiroya Fujisaki, Hidemichi Kouzu, Katsuhiko Ohori, Masaki Katayose, Akiyoshi Hashimoto, Masato Furuhashi

The Journal of nutrition, health and aging ( Elsevier BV ) 29 ( 1 ) 100434 - 100434 2025.01

The serum γ-Glutamyltransferase Level is Associated with the Development of Hypertension in Alcohol Infrequent Drinkers but not in Frequent Drinkers

Kazuma Mori, Marenao Tanaka, Tatsuya Sato, Yukinori Akiyama, Hirofumi Ohnishi, Nagisa Hanawa, Masato Furuhashi

Internal Medicine ( Japanese Society of Internal Medicine ) 2025

Urinary fatty acid‐binding protein 4 is a promising biomarker for glomerular damage in patients with diabetes mellitus

Marenao Tanaka, Tatsuya Sato, Tomohito Gohda, Nozomu Kamei, Maki Murakoshi, Erika Ishiwata, Keisuke Endo, Wataru Kawaharata, Hiroki Aida, Kei Nakata, Yukinori Akiyama, Mitsunobu Kubota, Michiyoshi Sanuki, Toru Suzuki, Yusuke Suzuki, Masato Furuhashi

Journal of Diabetes Investigation ( Wiley ) 2024.12

ABSTRACT Aims/Introduction Fatty acid‐binding protein (FABP) 4, which acts as an adipokine secreted by adipocytes, macrophages, and capillary endothelial cells, is expressed in injured glomerular cells. It has been reported that urinary (U‐) FABP4 is associated with renal dysfunction and proteinuria in several glomerular kidney diseases. However, the clinical significance of U‐FABP4 in diabetic kidney disease (DKD) remains undetermined. Materials and Methods Immunohistological analyses of FABP4 and FABP1 (liver‐type FABP), an established biomarker for impaired proximal tubules, were performed in the kidneys of patients with DKD and nonobese diabetic mice (KK‐Ta/Akita mice). The associations between U‐FABP4 and U‐FABP1 with kidney function and metabolic indices were also investigated in patients with type 1 diabetes (n = 57, mean age: 61 years) and patients with type 2 diabetes (n = 608, mean age: 65 years). Results In both patients with diabetes and diabetic mice, FABP4 was expressed in injured glomeruli with increased markers of endoplasmic reticulum stress in addition to peritubular capillaries, whereas FABP1 was mainly expressed in proximal tubules. Levels of U‐FABP4 and U‐FABP1 were independently associated with each other, and both levels were independently associated with estimated glomerular filtration rate (eGFR) and urinary albumin‐to‐creatinine ratio (UACR) after adjustment of age, sex, type of diabetes, duration of diabetes, and systolic blood pressure in patients with diabetes. Conclusions Urinary level of FABP4 derived from injured glomeruli with increased endoplasmic reticulum stress is independently associated with eGFR and UACR, suggesting a promising biomarker for glomerular damage in patients with diabetes.

A silent interplay between elevated intraocular pressure, glaucoma, and hypertension.

Tatsuya Sato, Araya Umetsu, Marenao Tanaka, Hiroshi Ohguro, Masato Furuhashi

Hypertension research : official journal of the Japanese Society of Hypertension 2024.12 [International journal]

セマグルチド注射薬により20年間のインスリン強化療法から離脱できた2型糖尿病の1例

寺沢誠, 中田圭, 大久保武志, 小川俊史, 隅田健太郎, 佐藤達也, 神津英至, 矢野俊之, 古橋眞人

糖尿病 ( (一社)日本糖尿病学会 ) 67 ( 12 ) 511 - 511 2024.12

腹腔鏡下胃スリーブ状切除術によりインスリン抵抗性-インスリン必要量増加-体重増加の悪循環を脱し、肥満関連合併症の改善が得られた1例

小川俊史, 佐藤達也, 大久保武志, 寺沢誠, 中田圭, 隅田健太郎, 神津英至, 矢野俊之, 矢島諒人, 伊東竜哉, 竹政伊知朗, 古橋眞人

日本内分泌学会雑誌 ( (一社)日本内分泌学会 ) 100 ( 4 ) 805 - 805 2024.12

LINC02154 promotes cell cycle and mitochondrial function in oral squamous cell carcinoma.

Takeshi Niinuma, Hiroshi Kitajima, Tatsuya Sato, Toshifumi Ogawa, Kazuya Ishiguro, Masahiro Kai, Eiichiro Yamamoto, Yui Hatanaka, Iyori Nojima, Mutsumi Toyota, Akira Yorozu, Shohei Sekiguchi, Noritsugu Tohse, Masato Furuhashi, Hiroshi Ohguro, Akihiro Miyazaki, Hiromu Suzuki

Cancer science 2024.11 [International journal]

Long noncoding RNAs (lncRNAs) play pivotal roles in the development of human malignancies, though their involvement in oral squamous cell carcinoma (OSCC) remains incompletely understood. Using The Cancer Genome Atlas (TCGA) dataset, we analyzed expression of 7840 lncRNAs in primary head and neck squamous cell carcinoma (HNSCC) and found that upregulation of LINC02154 is associated with a poorer prognosis. LINC02154 knockdown in OSCC cell lines induced cell cycle arrest and apoptosis, and significantly attenuated tumor growth in vitro and in vivo. Notably, depletion of LINC02154 downregulated FOXM1, a master regulator of cell cycle-related genes. RNA pulldown and mass spectrometry analyses identified a series of proteins that could potentially interact with LINC02154, including HNRNPK and LRPPRC. HNRNPK stabilizes FOXM1 expression by interacting with the 3'-UTR of FOXM1 mRNA, which suggests LINC02154 and HNRNPK promote cell cycling by regulating FOXM1 expression. Additionally, LINC02154 positively regulates HNRNPK expression by inhibiting microRNAs targeting HNRPNK. Moreover, LINC02154 affects mitochondrial function by interacting with LRPPRC. Depletion of LINC02154 suppressed expression of mitochondrial genes, including MTCO1 and MTCO2, and inhibited mitochondrial respiratory function in OSCC cells. These results suggest that LINC02154 exerts its oncogenic effects by modulating the cell cycle and oxidative phosphorylation in OSCC, highlighting LINC02154 as a potential therapeutic target.

mTOR Inhibitors Modulate the Biological Nature of TGF-β2-Treated or -Untreated Human Trabecular Meshwork Cells in Different Manners.

Megumi Watanabe, Tatsuya Sato, Toshiyuki Yano, Megumi Higashide, Toshifumi Ogawa, Nami Nishikiori, Masato Furuhashi, Hiroshi Ohguro

Biomedicines 12 ( 11 ) 2024.11 [International journal]

Background/Objectives: Mammalian target of rapamycin (mTOR) inhibition may have been suggested to have a beneficial effect on the glaucomatous human trabecular meshwork (HTM). To study the effects of the mTOR inhibitors rapamycin (Rapa) and Torin1 on the glaucomatous HTM, transforming growth factor-β2 (TGF-β2)-treated two-dimensionally (2D) and three-dimensionally (3D) cultured HTM cells were used. Methods: We evaluated (1) the levels of autophagy via Western blot analysis using a specific antibody against microtubule-associated protein 1 light chain 3 (LC3), (2) barrier capacity based on transepithelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC) permeability (2D), (3) cellular metabolic functions (2D), (4) the size and stiffness of spheroids, and (5) the mRNA expression of ECM proteins. Results: TGF-β2-induced inhibition of autophagy was significantly inhibited by Rapa and Torin1. Rapa and Torin1 substantially decreased barrier capacity in both TGF-β2-untreated and TGF-β2-treated HTM cells. Cellular metabolic analysis indicated that Rapa, but not Torin1, substantially enhanced both mitochondrial and glycolytic functions of TGF-β2-untreated HTM cells. In the physical properties of spheroids, TGF-β2 resulted in the formation of down-sized and stiffened spheroids. mTOR inhibitors decreased the size but not the stiffness of TGF-β2-untreated spheroids and significantly reduced the TGF-β2-related increase in the stiffness but not the size of spheroids. The diverse effects of mTOR inhibitors on TGF-β2-untreated and TGF-β2-treated spheroids were also observed in the mRNA expression of extracellular matrix proteins. Conclusions: The results taken together suggest that mTOR inhibitors significantly influence the biological aspects of both a single layer and multiple layers of the TGF-β2-treated HTM and untreated HTM.

TGF-β effects on adipogenesis of 3T3-L1 cells differ in 2D and 3D cell culture conditions.

Araya Umetsu, Megumi Watanabe, Tatsuya Sato, Megumi Higashide, Nami Nishikiori, Masato Furuhashi, Hiroshi Ohguro

FEBS open bio 2024.10 [International journal]

The TGF-β superfamily plays a pivotal role in the regulation of adipogenesis, but little is known about the potential differential role of the three isoforms of TGF-β, TGF-β-1~3. To further elucidate their role, two-dimensionally (2D) and three-dimensionally (3D) cultured 3T3-L1 mouse preadipocytes were subjected to the following analyses: (a) qPCR analysis of adipogenesis-related factors and major extracellular matrix protein (2D and /or 3D), (b) lipid staining by Oil Red O (2D) or BODIPY (3D), (c) Seahorse cellular metabolic measurement (2D), and (d) size and stiffness measurements of 3D 3T3-L1 spheroids. In the 2D cultured 3T3-L1 cells, mRNA expression levels of adipogenesis-related genes and Oil Red O lipid staining intensity were significantly increased by adipogenesis and they were substantially decreased following treatment with 0.1 nm TGF-β isoforms, with TGF-β2 having the greater effects. Consistent with these results, treatment with TGF-β2 resulted in suppression of mitochondrial and glycolytic functions in 2D cultured 3T3-L1 cells. However, the inhibitory effect of TGF-β on adipogenesis decreased under 3D spheroid culture conditions and TGF-β isoforms did not affect adipogenesis-induced (a) enlargement and downsizing of 3T3-L1 spheroids, (b) increase in BODIPY lipid staining intensity, and (c) up-regulation of the mRNA expression of adipogenesis-related genes. The findings presented herein suggest that the three TGF-β isoforms have different suppressive effects on adipogenesis-related cellular properties of 2D cultured 3T3-L1 cells and that their effects decrease under 3D spheroid culture conditions.

AI活用の血圧および健康評価へ大規模健診データを基にした高血圧および糖尿病の新規発症を予測する機械学習/AIモデルの構築

田中希尚, 古橋眞人

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 46回 286 - 286 2024.10

腹部大動脈瘤拡大因子としての瘤壁三次リンパ組織形成および濾胞関連新規リンパ球サブセットの解明

保坂到, 池上一平, 三上拓真, 佐藤達也, 小川俊文, 武川慶, 田中希尚, 遠藤圭佑, 秋山幸功, 大川陽史, 仲澤順二, 柴田豪, 中島智博, 伊庭裕, 高野賢一, 一宮慎吾, 川原田修義, 古橋眞人

脈管学 ( (一社)日本脈管学会 ) 64 ( Suppl. ) S149 - S150 2024.10

脈管分野において人工知能が果たす役割 Graph based clusteringを用いた腹部大動脈瘤拡大因子としての濾胞関連新規リンパ球サブセットの同定と機能的意義の解明

保坂到, 池上一平, 三上拓真, 佐藤達也, 小川俊文, 田中希尚, 遠藤圭佑, 秋山幸功, 大川陽史, 仲澤順二, 柴田豪, 中島智博, 伊庭裕, 高野賢一, 一宮慎吾, 川原田修義, 古橋眞人

脈管学 ( (一社)日本脈管学会 ) 64 ( Suppl. ) S114 - S114 2024.10

Unexpected and Synergistical Effects of All-Trans Retinoic Acid and TGF-β2 on Biological Aspects of 2D and 3D Cultured ARPE19 Cells.

Megumi Higashide, Megumi Watanabe, Tatsuya Sato, Toshifumi Ogawa, Araya Umetsu, Soma Suzuki, Masato Furuhashi, Hiroshi Ohguro, Nami Nishikiori

Biomedicines 12 ( 10 ) 2024.09 [International journal]

Objectives: To study the effects of all-trans retinoic acid (ATRA) on TGF-β2-induced effects of human retinal pigment epithelium cells under normoxia and hypoxia conditions. Methods: Two-dimensionally (2D) and three-dimensionally (3D) cultured ARPE19 cells were subjected to cellular functional analyses by transepithelial electrical resistance (TEER) and an extracellular flux assay (2D), measurement of levels of reactive oxygen species (ROS), gene expression analyses of COL1, αSMA, Zo-1, HIF1α, and PGC1α (2D), and physical property analyses (3D). Results: Under a normoxia condition, treatment with 100 nM ATRA substantially decreased barrier function regardless of the presence of 5 ng/mL TGF-β2 in 2D ARPE19 monolayer cells. Under a hypoxia condition, treatment with ATRA conversely increased barrier function, but the effect was masked by a marked increase in effects induced by TGF-β2. Although ATRA alone did not affect cellular metabolism and ROS levels in 2D ARPE cells, treatment with ATRA under a hypoxia condition did not affect ROS levels but shifted cellular metabolism from mitochondrial respiration to glycolysis. The changes of cellular metabolism and ROS levels were more pronounced with treatment of both ATRA and TGF-β2 independently of oxygen conditions. Changes in mRNA expressions of some of the above genes suggested the involvement of synergistical regulation of cellular functions by TGF-β2 and hypoxia. In 3D ARPE spheroids, the size was decreased and the stiffness was increased by either treatment with TGF-β2 or ATRA, but these changes were unexpectedly modulated by both ATRA and TGF-β2 treatment regardless of oxygen conditions. Conclusions: The findings reported herein indicate that TGF-β2 and hypoxia synergistically and differentially induce effects in 2D and 3D cultured ARPE19 cells and that their cellular properties are significantly altered by the presence of ATRA.

Application of Single Cell Type-Derived Spheroids Generated by Using a Hanging Drop Culture Technique in Various In Vitro Disease Models: A Narrow Review.

Hiroshi Ohguro, Megumi Watanabe, Tatsuya Sato, Nami Nishikiori, Araya Umetsu, Megumi Higashide, Toshiyuki Yano, Hiromu Suzuki, Akihiro Miyazaki, Kohichi Takada, Hisashi Uhara, Masato Furuhashi, Fumihito Hikage

Cells 13 ( 18 ) 2024.09 [International journal]

Cell culture methods are indispensable strategies for studies in biological sciences and for drug discovery and testing. Most cell cultures have been developed using two-dimensional (2D) culture methods, but three-dimensional (3D) culture techniques enable the establishment of in vitro models that replicate various pathogenic conditions and they provide valuable insights into the pathophysiology of various diseases as well as more precise results in tests for drug efficacy. However, one difficulty in the use of 3D cultures is selection of the appropriate 3D cell culture technique for the study purpose among the various techniques ranging from the simplest single cell type-derived spheroid culture to the more sophisticated organoid cultures. In the simplest single cell type-derived spheroid cultures, there are also various scaffold-assisted methods such as hydrogel-assisted cultures, biofilm-assisted cultures, particle-assisted cultures, and magnet particle-assisted cultures, as well as non-assisted methods, such as static suspension cultures, floating cultures, and hanging drop cultures. Since each method can be differently influenced by various factors such as gravity force, buoyant force, centrifugal force, and magnetic force, in addition to non-physiological scaffolds, each method has its own advantages and disadvantages, and the methods have different suitable applications. We have been focusing on the use of a hanging drop culture method for modeling various non-cancerous and cancerous diseases because this technique is affected only by gravity force and buoyant force and is thus the simplest method among the various single cell type-derived spheroid culture methods. We have found that the biological natures of spheroids generated even by the simplest method of hanging drop cultures are completely different from those of 2D cultured cells. In this review, we focus on the biological aspects of single cell type-derived spheroid culture and its applications in in vitro models for various diseases.

Contribution of MLKL to the development of doxorubicin-induced cardiomyopathy and its amelioration by rapamycin.

Masaki Shimizu, Wataru Ohwada, Toshiyuki Yano, Hidemichi Kouzu, Tatsuya Sato, Toshifumi Ogawa, Arata Osanami, Yuki Toda, Hiroshi Nagahama, Masaya Tanno, Tetsuji Miura, Atsushi Kuno, Masato Furuhashi

Journal of pharmacological sciences 156 ( 1 ) 9 - 18 2024.09 [Domestic journal]

Necroptosis, necrosis characterized by RIPK3-MLKL activation, has been proposed as a mechanism of doxorubicin (DOX)-induced cardiomyopathy. We showed that rapamycin, an mTORC1 inhibitor, attenuates cardiomyocyte necroptosis. Here we examined role of MLKL in DOX-induced myocardial damage and protective effects of rapamycin. Cardiomyopathy was induced in mice by intraperitoneal injections of DOX (10 mg/kg, every other day) and followed for 7 days. DOX-treated mice showed a significant decline in LVEF assessed by cardiac MRI (45.5 ± 5.1% vs. 65.4 ± 4.2%), reduction in overall survival rates, and increases in myocardial RIPK3 and MLKL expression compared with those in vehicle-treated mice, and those changes were prevented by administration of rapamycin (0.25 mg/kg) before DOX injection. In immunohistochemical analyses, p-MLKL signals were detected in the cardiomyocytes of DOX-treated mice, and the signals were reduced by rapamycin. Mlkl+/- and Mlkl-/- mice were similarly resistant to DOX-induced cardiac dysfunction, indicating that a modest reduction in MLKL level is sufficient to prevent the development of DOX-induced cardiomyopathy. However, evidence of cardiomyocyte necrosis assessed by C9 immunostaining, presence of replacement fibrosis, and electron microscopic analyses was negligible in the myocardium of DOX-treated mice. Thus, MLKL-mediated signaling contributes to DOX-induced cardiac dysfunction primarily by a necrosis-independent mechanism, which is inhibitable by rapamycin.

FABP5 Is a Possible Factor for the Maintenance of Functions of Human Non-Pigmented Ciliary Epithelium Cells.

Megumi Higashide, Megumi Watanabe, Tatsuya Sato, Araya Umetsu, Nami Nishikiori, Toshifumi Ogawa, Masato Furuhashi, Hiroshi Ohguro

International journal of molecular sciences 25 ( 17 ) 2024.08 [International journal]

To elucidate the possible biological roles of fatty acid-binding protein 5 (FABP5) in the intraocular environment, the cells from which FABP5 originates were determined by using four different intraocular tissue-derived cell types including human non-pigmented ciliary epithelium (HNPCE) cells, retinoblastoma (RB) cells, adult retinal pigment epithelial19 (ARPE19) cells and human ocular choroidal fibroblast (HOCF) cell lines, and the effects of FABP ligand 6, a specific inhibitor for FABP5 and FABP7 were analyzed by RNA sequencing and seahorse cellular metabolic measurements. Among these four different cell types, qPCR analysis showed that FABP5 was most prominently expressed in HNPCE cells, in which no mRNA expression of FABP7 was detected. In RNA sequencing analysis, 166 markedly up-regulated and 198 markedly down-regulated differentially expressed genes (DEGs) were detected between non-treated cells and cells treated with FABP ligand 6. IPA analysis of these DEGs suggested that FABP5 may be involved in essential roles required for cell development, cell survival and cell homeostasis. In support of this possibility, both mitochondrial and glycolytic functions of HNPCE cells, in which mRNA expression of FABP5, but not that of FABP7, was detected, were shown by using a Seahorse XFe96 Bioanalyzer to be dramatically suppressed by FABP ligand 6-induced inhibition of the activity of FABP5. Furthermore, in IPA upstream analysis, various unfolded protein response (UPR)-related factors were identified as upstream and causal network master regulators. Analysis by qPCR analysis showed significant upregulation of the mRNA expression of most of UPR-related factors and aquaporin1 (AQP1). The findings in this study suggest that HNPCE is one of intraocular cells producing FABP5 and may be involved in the maintenance of UPR and AQP1-related functions of HNPCE.

Straightforward and immediate ultrasound-guided kidney biopsy using a guide needle technique to get adequate tissue with reduced procedural time.

Hiroaki Komatsu, Tomohisa Yamashita, Arata Osanami, Chikako Akazawa, Kota Endo, Shun Tsugawa, Ayumu Kimura, Daisuke Miyamori, Koki Abe, Satoko Takahashi, Yufu Gocho, Masayuki Koyama, Tatsuya Sato, Marenao Tanaka, Norihito Moniwa, Masato Furuhashi

Clinical and experimental nephrology 2024.08 [Domestic journal]

BACKGROUND: A streamlined and effective renal biopsy technique is essential for all nephrologists, particularly those who are less experienced, such as residents. Herein, we report the efficacy of a Straightforward and Immediate ultrasound-guided kidney biopsy using a Guide Needle (SIGN) technique, which allows operators to insert a biopsy gun through a guide needle placed into the fascia of the posterior abdominal wall. METHODS: A retrospective cross-sectional study was conducted at a nephrology training institution to compare the time spent on the procedure and the number of glomeruli obtained between a group using the SIGN (n = 81) and a group using the conventional ultrasound-guided kidney biopsy technique with a needle guide device (n = 143). RESULTS: The median procedure time in the SIGN group (2 min, interquartile range [IQR]: 1-3 min) was significantly shorter than that in the conventional group (3 min, IQR: 2-4 min) (P < 0.001). Multivariable linear regression and logistic regression analyses adjusted for covariates, including operators (board-certificated nephrologists or nephrology residents), showed that the use of the SIGN technique was independently associated with a high number of glomeruli obtained and a procedure time above 2 min as the median value (odds ratio: 0.17, 95% confidence interval CI 0.09-0.34). The prevalence of complications was comparable between the two groups (P = 0.681). CONCLUSION: The SIGN technique reduces the procedure time and obtains adequate biopsy tissue regardless of the operator's experience. SIGN can be applied in nephrology training programs and used as a standard biopsy technique.

Intraocular fatty acids induce reinforcement of barrier functions on the outer blood-retinal barrier.

Nami Nishikiori, Megumi Watanabe, Tatsuya Sato, Araya Umetsu, Megumi Higashide, Masato Furuhashi, Hiroshi Ohguro

Prostaglandins, leukotrienes, and essential fatty acids 202 102637 - 102637 2024.08 [International journal]

The aim of the present study was to elucidate unknown effects of intraocular fatty acids (ioFAs) including palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1), linoleic acid (C18:2), arachidonic acid (C20:4), eicosapentaenoic acid (EPA, C20:5) and docosahexaenoic acid (DHA, C22:6) on the outer blood-retinal barrier (oBRB). For this purpose, human retinal pigment epithelium cell line ARPE19 was subjected to analyses for evaluating the following biological phenotypes: (1) cell viability, (2) cellular metabolic functions, (3) barrier functions by trans-epithelial electrical resistance (TEER), and (4) expression of tight junction (TJ) molecules. In the presence of 100 nM ioFAs, no significant effects on cell viability of ARPE19 cells was observed. While treatment with EPA or DHA tended to reduce non-mitochondrial oxygen consumption, most indices in mitochondrial functions were not markedly affected by treatment with ioFAs in ARPE19 cells. On the other hand, ioFAs except for palmitic acid and stearic acid significantly increased basal extracellular acidification rates, suggesting activated glycolysis or increased lactate production. Interestingly, TEER values of planar ARPE19 monolayer were significantly increased by treatment any ioFAs. Consistently, gene expression levels of TJ proteins were increased by treatment with ioFAs. Collectively, the findings presented herein suggest that ioFAs may contribute to reinforcement of barrier functions of the oBRB albeit there are some differences in biological effects depending on the type of ioFAs.

Metabolic dysfunction-associated steatotic liver disease (SLD) and alcohol-associated liver disease, but not SLD without metabolic dysfunction, are independently associated with new onset of chronic kidney disease during a 10-year follow-up period.

Kazuma Mori, Marenao Tanaka, Tatsuya Sato, Yukinori Akiyama, Keisuke Endo, Toshifumi Ogawa, Toru Suzuki, Hiroki Aida, Wataru Kawaharata, Kei Nakata, Itaru Hosaka, Araya Umetsu, Nagisa Hanawa, Masato Furuhashi

Hepatology research : the official journal of the Japan Society of Hepatology 2024.08 [International journal]

AIMS: The new nomenclature of steatotic liver disease (SLD) including metabolic dysfunction-associated SLD (MASLD), MASLD and increased alcohol intake (MetALD), and alcohol-associated liver disease (ALD) has recently been proposed. We aimed to elucidate the relationship between each category of SLD and chronic kidney disease (CKD). METHODS: We investigated the effects of various SLDs on the development of CKD, defined as estimated glomerular filtration rate (eGFR) <60 mL/min/1.73 m2 or positive for urinary protein, during a 10-year period in 12 138 Japanese subjects (men / women, 7984/4154; mean age, 48 years) who received annual health examinations including abdominal ultrasonography. RESULTS: The prevalences of SLD without metabolic dysfunction (SLD-MD[-]), MASLD, MetALD, and ALD were 1.7%, 26.3%, 4.9%, and 1.9%, respectively. During the follow-up period, 1963 subjects (16.2%) (men / women, 1374 [17.2%]/589 [14.2%]) had new onset of CKD. Multivariable Cox proportional hazard model analyses after adjustment of age, sex, eGFR, current smoking habit, diabetes mellitus, hypertension, and dyslipidemia showed that the hazard ratios (HR [95% confidence interval]) for the development of CKD in subjects with MASLD (1.20 [1.08-1.33], p = 0.001) and those with ALD (1.41 [1.05-1.88], p = 0.022), but not those with MetALD (1.11 [0.90-1.36], p = 0.332), were significantly higher than the HR in subjects with non-SLD. Interestingly, subjects with SLD-MD[-] had a significantly lower HR (0.61 [0.39-0.96], p = 0.034) than that in subjects with non-SLD. The addition of the novel classification of SLDs into traditional risk factors for the development of CKD significantly improved the discriminatory capacity. CONCLUSIONS: MASLD and ALD, but not SLD-MD[-], are independently associated with the development of CKD.

Associations between circulating levels of FABP4 and TNF receptors are more evident in patients with type 2 diabetes mellitus than in patients with type 1 diabetes mellitus.

Marenao Tanaka, Tomohito Gohda, Nozomu Kamei, Maki Murakoshi, Tatsuya Sato, Mitsunobu Kubota, Michiyoshi Sanuki, Erika Ishiwata, Keisuke Endo, Yusuke Suzuki, Masato Furuhashi

Endocrine connections 2024.08 [International journal]

BACKGROUND: Fatty acid-binding protein 4 (FABP4) is an adipokine that plays significant roles in the development of insulin resistance and atherosclerosis. High levels of soluble tumor necrosis factor receptors (TNFRs) including TNFR1 and TNFR2 are associated with renal dysfunction and increased mortality in patients with diabetes mellitus (DM). However, the association between circulating levels of FABP4 and TNFRs remains unclear. METHODS: We investigated the associations of FABP4 with TNFRs and metabolic markers in Japanese patients with type 1 DM (T1DM, n=76, men/women: 31/45) and type 2 DM (T2DM, n=575, men/women: 312/263). RESULTS: FABP4 concentration was positively correlated with levels of TNFR1 and TNFR2 in both patients with T1DM and those with T2DM. Multivariable regression analyses showed that there were independent associations of FABP4 concentration with body mass index (BMI) and estimated glomerular filtration rate (eGFR) after adjustment of age and sex in both patients with T1DM and those with T2DM. FABP4 concentration was independently associated with circulating levels of TNFR1 and TNFR2 after adjustment of the confounders in patients with T2DM but not in those with T1DM. Similarly, levels of TNFR1 and TNFR2 were independently associated with FABP4 concentration after adjustment of age, sex, systolic blood pressure, duration of DM and levels of eGFR, high-density lipoprotein cholesterol and C-reactive protein in patients with T2DM but not in those with T1DM. CONCLUSION: FABP4 concentration is independently associated with levels of TNFRs in patients with DM, but the association is more evident in patients with T2DM than in those with T1DM.

High Intraocular Pressure Is Independently Associated With New-Onset Systemic Hypertension Over a 10-Year Period

Araya Umetsu, Marenao Tanaka, Tatsuya Sato, Yukinori Akiyama, Keisuke Endo, Kazuma Mori, Hirofumi Ohnishi, Megumi Watanabe, Hiroshi Ohguro, Nagisa Hanawa, Masato Furuhashi

Circulation Journal ( Japanese Circulation Society ) 2024.07

Fatty Acid-Binding Protein 4-Mediated Regulation Is Pivotally Involved in Retinal Pathophysiology: A Review.

Hiroshi Ohguro, Megumi Watanabe, Fumihito Hikage, Tatsuya Sato, Nami Nishikiori, Araya Umetsu, Megumi Higashide, Toshifumi Ogawa, Masato Furuhashi

International journal of molecular sciences 25 ( 14 ) 2024.07 [International journal]

Fatty acid-binding proteins (FABPs), a family of lipid chaperone molecules that are involved in intracellular lipid transportation to specific cellular compartments, stimulate lipid-associated responses such as biological signaling, membrane synthesis, transcriptional regulation, and lipid synthesis. Previous studies have shown that FABP4, a member of this family of proteins that are expressed in adipocytes and macrophages, plays pivotal roles in the pathogenesis of various cardiovascular and metabolic diseases, including diabetes mellitus (DM) and hypertension (HT). Since significant increases in the serum levels of FABP4 were detected in those patients, FABP4 has been identified as a crucial biomarker for these systemic diseases. In addition, in the field of ophthalmology, our group found that intraocular levels of FABP4 (ioFABP4) and free fatty acids (ioFFA) were substantially elevated in patients with retinal vascular diseases (RVDs) including proliferative diabetic retinopathy (PDR) and retinal vein occlusion (RVO), for which DM and HT are also recognized as significant risk factors. Recent studies have also revealed that ioFABP4 plays important roles in both retinal physiology and pathogenesis, and the results of these studies have suggested potential molecular targets for retinal diseases that might lead to future new therapeutic strategies.

Lysophosphatidic Acid Modulates TGF-β2-Induced Biological Phenotype in Human Conjunctival Fibroblasts.

Megumi Watanabe, Yuri Tsugeno, Tatsuya Sato, Megumi Higashide, Nami Nishikiori, Araya Umetsu, Toshifumi Ogawa, Masato Furuhashi, Hiroshi Ohguro

Life (Basel, Switzerland) 14 ( 6 ) 2024.06 [International journal]

BACKGROUND: Although lysophosphatidic acid (LPA) is known to have multiple pathophysiological roles, its contributions to ocular tissues, especially conjunctival fibrogenesis, remain to be elucidated. METHODS: To study this issue, the effects of LPA on transforming growth factor-β2 (TGF-β2)-induced fibrogenesis of two-dimensional (2D) and three-dimensional (3D) cultures of human conjunctival fibroblasts (HconF) were examined by the following analyses: (1) planar proliferation determined by transepithelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC)-dextran permeability measurements, (2) real-time metabolic analyses, (3) measurements of the size and stiffness of 3D spheroids, and (4) mRNA expression of extracellular matrix (ECM) molecules and their modulators. RESULTS: LPA had no effect on TGF-β2-induced increase in the planar proliferation of HconF cells. LPA induced a more quiescent metabolic state in 2D HconF cells, but this metabolic suppression by LPA was partially blunted in the presence of TGF-β2. In contrast, LPA caused a substantial decrease in the hardness of 3D HconF spheroids independently of TGF-β2. In agreement with these different LPA-induced effects between 2D and 3D cultured HconF cells, mRNA expressions of ECM and their modulators were differently modulated. CONCLUSION: The findings that LPA induced the inhibition of both TGF-β2-related and -unrelated subepithelial proliferation of HconF cells may be clinically applicable.

TGF-β Isoforms and Local Environments Greatly Modulate Biological Nature of Human Retinal Pigment Epithelium Cells.

Nami Nishikiori, Tatsuya Sato, Toshifumi Ogawa, Megumi Higashide, Araya Umetsu, Soma Suzuki, Masato Furuhashi, Hiroshi Ohguro, Megumi Watanabe

Bioengineering (Basel, Switzerland) 11 ( 6 ) 2024.06 [International journal]

To characterize transforming growth factor-β (TGF-β) isoform (TGF-β1~3)-b's biological effects on the human retinal pigment epithelium (RPE) under normoxia and hypoxia conditions, ARPE19 cells cultured by 2D (two-dimensional) and 3D (three-dimensional) conditions were subjected to various analyses, including (1) an analysis of barrier function by trans-epithelial electrical resistance (TEER) measurements; (2) qPCR analysis of major ECM molecules including collagen 1 (COL1), COL4, and COL6; α-smooth muscle actin (αSMA); hypoxia-inducible factor 1α (HIF1α); and peroxisome proliferator-activated receptor-gamma coactivator (PGC1α), a master regulator for mitochondrial respiration;, tight junction-related molecules, Zonula occludens-1 (ZO1) and E-cadherin; and vascular endothelial growth factor (VEGF); (3) physical property measurements of 3D spheroids; and (4) cellular metabolic analysis. Diverse effects among TGF-β isoforms were observed, and those effects were also different between normoxia and hypoxia conditions: (1) TGF-β1 and TGF-β3 caused a marked increase in TEER values, and TGF-β2 caused a substantial increase in TEER values under normoxia conditions and hypoxia conditions, respectively; (2) the results of qPCR analysis supported data obtained by TEER; (3) 3D spheroid sizes were decreased by TGF-β isoforms, among which TGF-β1 had the most potent effect under both oxygen conditions; (4) 3D spheroid stiffness was increased by TGF-β2 and TGF-β3 or by TGF-β1 and TGF-β3 under normoxia conditions and hypoxia conditions, respectively; and (5) the TGF-β isoform altered mitochondrial and glycolytic functions differently under oxygen conditions and/or culture conditions. These collective findings indicate that the TGF-β-induced biological effects of 2D and 3D cultures of ARPE19 cells were substantially diverse depending on the three TGF-β isoforms and oxygen levels, suggesting that pathological conditions including epithelial-mesenchymal transition (EMT) of the RPE may be exclusively modulated by both factors.

FABP4 Is an Indispensable Factor for Regulating Cellular Metabolic Functions of the Human Retinal Choroid.

Hiroshi Ohguro, Megumi Watanabe, Tatsuya Sato, Nami Nishikiori, Araya Umetsu, Megumi Higashide, Toshifumi Ogawa, Masato Furuhashi

Bioengineering (Basel, Switzerland) 11 ( 6 ) 2024.06 [International journal]

The purpose of the current study was to elucidate the physiological roles of intraocularly present fatty acid-binding protein 4 (FABP4). Using four representative intraocular tissue-derived cell types, including human non-pigmented ciliary epithelium (HNPCE) cells, retinoblastoma (RB) cells, adult retinal pigment epithelial19 (ARPE19) cells and human ocular choroidal fibroblast (HOCF) cells, the intraocular origins of FABP4 were determined by qPCR analysis, and the intracellular functions of FABP4 were investigated by seahorse cellular metabolic measurements and RNA sequencing analysis using a specific inhibitor for FABP4, BMS309403. Among these four different cell types, FABP4 was exclusively expressed in HOCF cells. In HOCF cells, both mitochondrial and glycolytic functions were significantly decreased to trace levels by BMS309403 in a dose-dependent manner. In the RNA sequencing analysis, 67 substantially up-regulated and 94 significantly down-regulated differentially expressed genes (DEGs) were identified in HOCF cells treated with BMS309403 and those not treated with BMS309403. The results of Gene Ontology enrichment analysis and ingenuity pathway analysis (IPA) revealed that the DEGs were most likely involved in G-alpha (i) signaling, cAMP-response element-binding protein (CREB) signaling in neurons, the S100 family signaling pathway, visual phototransduction and adrenergic receptor signaling. Furthermore, upstream analysis using IPA suggested that NKX2-1 (thyroid transcription factor1), HOXA10 (homeobox A10), GATA2 (gata2 protein), and CCAAT enhancer-binding protein A (CEBPA) were upstream regulators and that NKX homeobox-1 (NKX2-1), SFRP1 (Secreted frizzled-related protein 1) and TREM2 (triggering receptor expressed on myeloid cells 2) were causal network master regulators. The findings in this study suggest that intraocularly present FABP4 originates from the ocular choroid and may be a critical regulator for the cellular homeostasis of non-adipocyte HOCF cells.

慢性腎臓病と代謝異常関連脂肪肝疾患(MAFLD)との合併は虚血性心疾患の発症リスクを増加させる

宮森大輔, 田中希尚, 佐藤達也, 遠藤圭佑, 森和真, 塙なぎさ, 大西浩文, 古橋眞人

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 66 ( 4 ) 652 - 652 2024.06

慢性腎臓病と代謝異常関連脂肪肝疾患(MAFLD)との合併は虚血性心疾患の発症リスクを増加させる

宮森大輔, 田中希尚, 佐藤達也, 遠藤圭佑, 森和真, 塙なぎさ, 大西浩文, 古橋眞人

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 66 ( 4 ) 652 - 652 2024.06

日本人2型糖尿病患者においてペマフィブラートの12ヵ月投与が血中脂質と脂肪肝指数に与える影響の検討

鈴木亨, 佐藤達也, 田中希尚, 遠藤圭佑, 中田圭, 小川俊史, 保坂致, 秋山幸功, 梅津新矢, 古橋眞人

日本動脈硬化学会総会プログラム・抄録集 ( (一社)日本動脈硬化学会 ) 56回 231 - 231 2024.06

Adverse plasma branched‐chain amino acid profile mirrors fatty muscle degeneration in diabetic heart failure patients

Hidemichi Kouzu, Toshiyuki Yano, Satoshi Katano, Wataru Kawaharata, Keishi Ogura, Ryo Numazawa, Ryohei Nagaoka, Katsuhiko Ohori, Ryo Nishikawa, Wataru Ohwada, Takefumi Fujito, Nobutaka Nagano, Masato Furuhashi

ESC Heart Failure ( Wiley ) 2024.05

Abstract Aims Elevated plasma branched‐chain amino acids (BCAAs) are tightly linked to incident diabetes and its complications, while lower BCAAs are associated with adverse outcomes in the elderly and heart failure (HF) patients. The interplay between body compositions and plasma BCAAs, especially under the influence of co‐morbid diabetes in HF patients, is not well understood. Here, we examined the impact of diabetes on the prognostic value of plasma BCAA and its association with body compositions in HF patients. Methods and results We retrospectively examined 301 HF patients (70 ± 15 years old; 59% male), among which 36% had diabetes. Blood samples for plasma BCAA measurements were collected in a fasting state after stabilization of HF and analysed using ultraperformance liquid chromatography. A dual‐energy X‐ray absorptiometry scan assessed regional body compositions, and muscle wasting was defined as appendicular skeletal muscle mass index (ASMI) < 7.00 and <5.40 kg/m² for males and females, respectively, according to the criteria of the Asian Working Group for Sarcopenia. Although analyses of covariance revealed that plasma BCAAs were significantly higher in diabetic patients, low valine (<222.1 nmol/mL) similarly predicted adverse events defined by HF hospitalization, lethal arrhythmia, or all‐cause death in both diabetic and non‐diabetic patients independently of age, sex, and NT‐proBNP (adjusted hazard ratio [HR] 3.1, 95% confidence interval [CI] of 1.1–8.6 and adjusted HR 2.67, 95% CI 1.1–6.5, respectively; P for interaction 0.88). In multivariate linear regression analyses comprising age, sex, and regional body compositions as explanatory variables, plasma BCAAs were positively correlated with visceral adipose tissue area in non‐diabetic patients (standardized β coefficients [β] = 0.44, P < 0.001). In contrast, in diabetic patients, plasma BCAAs were correlated positively with ASMI (β = 0.49, P = 0.001) and negatively with appendicular fat mass index (AFMI; β = −0.42, P = 0.004). Co‐morbid diabetes was independently associated with muscle wasting (adjusted odds ratio 2.1, 95% CI 1.1–4.0) and significantly higher plasma 3‐methylhistidine level, a marker of myofibrillar degradation. In diabetic patients, ASMI uniquely showed a J‐shaped relationship with AFMI, and in a subgroup of HF patients with muscle wasting, diabetic patients showed 12% higher AFMI than non‐diabetic patients despite comparable ASMI reductions. Conclusions Despite higher plasma BCAA levels in HF patients with diabetes, the prognostic value of low valine remained consistent regardless of diabetes status. However, low BCAAs were distinctly associated with fatty muscle degeneration in the extremities in diabetic patients, suggesting the importance of targeted interventions to prevent such tissue remodelling in this population.

The Specific ROCK2 Inhibitor KD025 Alleviates Glycolysis through Modulating STAT3-, CSTA- and S1PR3-Linked Signaling in Human Trabecular Meshwork Cells.

Megumi Watanabe, Tatsuya Sato, Araya Umetsu, Toshifumi Ogawa, Nami Nishikiori, Megumi Suzuki, Masato Furuhashi, Hiroshi Ohguro

Biomedicines 12 ( 6 ) 2024.05 [International journal]

To investigate the biological significance of Rho-associated coiled-coil-containing protein kinase (ROCK) 2 in the human trabecular meshwork (HTM), changes in both metabolic phenotype and gene expression patterns against a specific ROCK2 inhibitor KD025 were assessed in planar-cultured HTM cells. A seahorse real-time ATP rate assay revealed that administration of KD025 significantly suppressed glycolytic ATP production rate and increased mitochondrial ATP production rate in HTM cells. RNA sequencing analysis revealed that 380 down-regulated and 602 up-regulated differentially expressed genes (DEGs) were identified in HTM cells treated with KD025 compared with those that were untreated. Gene ontology analysis revealed that DEGs were more frequently related to the plasma membrane, extracellular components and integral cellular components among cellular components, and related to signaling receptor binding and activity and protein heterodimerization activity among molecular functions. Ingenuity Pathway Analysis (IPA) revealed that the detected DEGs were associated with basic cellular biological and physiological properties, including cellular movement, development, growth, proliferation, signaling and interaction, all of which are associated with cellular metabolism. Furthermore, the upstream regulator analysis and causal network analysis estimated IL-6, STAT3, CSTA and S1PR3 as possible regulators. Current findings herein indicate that ROCK2 mediates the IL-6/STAT3-, CSTA- and S1PR3-linked signaling related to basic biological activities such as glycolysis in HTM cells.

Associations between in‐hospital daily protein intake and adverse clinical outcomes in older patients with heart failure

Satoshi Katano, Toshiyuki Yano, Kotaro Yamano, Ryo Numazawa, Ryohei Nagaoka, Suguru Honma, Yusuke Fujisawa, Katsuhiko Ohori, Hidemichi Kouzu, Hayato Kunihara, Hiroya Fujisaki, Masaki Katayose, Akiyoshi Hashimoto, Masato Furuhashi

ESC Heart Failure ( Wiley ) 2024.05

Abstract Aims The adverse effects of low daily protein intake (DPI) on clinical outcomes in patients with heart failure (HF) are known; however, an optimal DPI to predict event adverse outcomes remains undetermined. Moreover, whether protein restriction therapy for chronic kidney disease is applicable in patients with HF and renal dysfunction remains unclear. Methods and results In this single‐centre, ambispective cohort study, we included 405 patients with HF aged ≥65 years (mean age, 78.6 ± 7.5 years; 50% women). DPI was estimated from consumption over three consecutive days before discharge and normalized relative to the ideal body weight [IBW, 22 kg/m² × height (m)²]. The primary outcome was a composite of all‐cause mortality and HF‐related readmission within the 2 year post‐discharge period. Results During an average follow‐up period of 1.49 ± 0.74 years, 100 patients experienced composite events. Kaplan–Meier survival curves revealed a significantly lower composite event‐free rate in patients within the lowest quartile of DPI than in the upper quartiles (log‐rank test, P = 0.02). A multivariate Cox proportional hazards analysis after adjusting for established prognostic markers and non‐proteogenic energy intake revealed that patients in the lowest DPI quartile faced a two‐fold higher risk of composite events than those in the highest quartile [hazard ratio (HR), 2.03; 95% confidence interval (CI), 1.08–3.82; P = 0.03]. The composite event risk linearly increased as DPI decreased (P for nonlinearity = 0.90), with each standard deviation (0.26 g/kg IBW/day) decrease in DPI associated with a 32% increase in composite event risk (HR, 1.32; 95% CI, 1.10–1.71; P = 0.04). There was significant heterogeneity in the effect of DPI, with the possible disadvantage of lower DPI in patients with HF with cystatin C‐based estimated glomerular filtration rate <30 mL/min/1.73 m². The cutoff value of DPI for predicting the occurrence of composite events calculated from the Youden index was 1.12 g/kg IBW/day. Incorporating a DPI < 1.12 g/kg IBW/day into the baseline model significantly improved the prediction of post‐discharge composite events (continuous net reclassification improvement, 0.294; 95% CI, 0.072–0.516; P = 0.01). Conclusions Lower DPI during hospitalization is associated with an increased risk of mortality and HF readmission independent of non‐proteogenic energy intake, and the possible optimal DPI for predicting adverse clinical outcomes is >1.12 g/kg IBW/day in older patients with HF. Caution is warranted when protein restriction therapy is administered to older patients with HF and renal dysfunction.

Modulation of Epithelial-Mesenchymal Transition Is a Possible Underlying Mechanism for Inducing Chemoresistance in MIA PaCa-2 Cells against Gemcitabine and Paclitaxel.

Hajime Nakamura, Megumi Watanabe, Kohichi Takada, Tatsuya Sato, Fumihito Hikage, Araya Umetsu, Joji Muramatsu, Masato Furuhashi, Hiroshi Ohguro

Biomedicines 12 ( 5 ) 2024.05 [International journal]

To elucidate the currently unknown molecular mechanisms responsible for the similarity and difference during the acquirement of resistance against gemcitabine (GEM) and paclitaxel (PTX) in patients with pancreatic carcinoma, we examined two-dimensional (2D) and three-dimensional (3D) cultures of parent MIA PaCa-2 cells (MIA PaCa-2-PA) and their GEM resistance cell line (MIA PaCa-2-GR) and PTX resistance (MIA PaCa-2-PR). Using these cells, we examined 3D spheroid configurations and cellular metabolism, including mitochondrial and glycolytic functions, with a Seahorse bio-analyzer and RNA sequencing analysis. Compared to the MIA PaCa-2-PA, (1) the formation of the 3D spheroids of MIA PaCa-2-GR or -PR was much slower, and (2) their mitochondrial and glycolytic functions were greatly modulated in MIA PaCa-2-GR or -PR, and such metabolic changes were also different between their 2D and 3D culture conditions. RNA sequencing and bioinformatic analyses of the differentially expressed genes (DEGs) using an ingenuity pathway analysis (IPA) suggested that various modulatory factors related to epithelial -mesenchymal transition (EMT) including STAT3, GLI1, ZNF367, NKX3-2, ZIC2, IFIT2, HEY1 and FBLX, may be the possible upstream regulators and/or causal network master regulators responsible for the acquirement of drug resistance in MIA PaCa-2-GR and -PR. In addition, among the prominently altered DEGs (Log2 fold changes more than 6 or less than -6), FABP5, IQSEC3, and GASK1B were identified as unique genes associated with their antisense RNA or pseudogenes, and among these, FABP5 and GASK1B are known to function as modulators of cancerous EMT. Therefore, the observations reported herein suggest that modulations of cancerous EMT may be key molecular mechanisms that are responsible for inducing chemoresistance against GEM or PTX in MIA PaCa-2 cells.

Tirzepatide ameliorates eating behaviors regardless of prior exposure to glucagon-like peptide receptor agonists in Japanese patients with type 2 diabetes mellitus

Toru Suzuki, Tatsuya Sato, Marenao Tanaka, Keisuke Endo, Kei Nakata, Toshifumi Ogawa, Itaru Hosaka, Yukinori Akiyama, Araya Umetsu, Masato Furuhashi

Journal of Diabetes and its Complications ( Elsevier BV ) 108779 - 108779 2024.05

Deciphering metabolic dysfunction-associated steatotic liver disease: insights from predictive modeling and clustering analysis.

Kazuma Mori, Yukinori Akiyama, Marenao Tanaka, Tatsuya Sato, Keisuke Endo, Itaru Hosaka, Nagisa Hanawa, Naoya Sakamoto, Masato Furuhashi

Journal of gastroenterology and hepatology 2024.04 [International journal]

BACKGROUND AND AIM: New nomenclature of steatotic liver disease (SLD) including metabolic dysfunction-associated SLD (MASLD), MASLD and increased alcohol intake (MetALD), and alcohol-associated liver disease (ALD) has recently been proposed. We investigated clustering analyses to decipher the complex landscape of SLD pathologies including the former nomenclature of nonalcoholic fatty liver disease (NAFLD) and metabolic dysfunction-associated fatty liver disease (MAFLD). METHODS: Japanese individuals who received annual health checkups including abdominal ultrasonography (n = 15 788, men/women: 10 250/5538, mean age: 49 years) were recruited. RESULTS: The numbers of individuals with SLD, MASLD, MetALD, ALD, NAFLD, and MAFLD were 5603 (35.5%), 4227 (26.8%), 795 (5.0%), 324 (2.1%), 3982 (25.8%), and 4946 (31.3%), respectively. Clustering analyses using t-distributed stochastic neighbor embedding and K-means to visually represent interconnections in SLDs uncovered five cluster formations. MASLD and NAFLD mainly shared three clusters including (i) low alcohol intake with relatively low-grade obesity; (ii) obesity with dyslipidemia; and (iii) dysfunction of glucose metabolism. Both MetALD and ALD displayed one distinct cluster intertwined with alcohol consumption. MAFLD widely shared all of the five clusters. In machine learning-based analyses using algorithms of random forest and extreme gradient boosting and receiver operating characteristic curve analyses, fatty liver index (FLI), calculated by body mass index, waist circumference, and levels of γ-glutamyl transferase and triglycerides, was selected as a useful feature for SLDs. CONCLUSIONS: The new nomenclature of SLDs is useful for obtaining a better understanding of liver pathologies and for providing valuable insights into predictive factors and the dynamic interplay of diseases. FLI may be a noninvasive predictive marker for detection of SLDs.

Which came first: Sarcopenia or weight loss?

Katsuhiko Ohori, Toshiyuki Yano, Satoshi Katano, Ryohei Nagaoka, Ryo Numazawa, Kotaro Yamano, Yusuke Fujisawa, Hidemichi Kouzu, Nobutaka Nagano, Takefumi Fujito, Ryo Nishikawa, Wataru Ohwada, Masato Furuhashi

Geriatrics & Gerontology International ( Wiley ) 2024.04

Sampsonの式あるいは人工知能を用いて推定したsmall dense LDLコレステロール値の精度の検証

大久保武志, 遠藤圭佑, 田中希尚, 佐藤達也, 小林亮, 田中真輝人, 秋山幸功, 保坂到, 中田圭, 小山雅之, 大西浩文, 高橋聡, 古橋眞人

糖尿病 ( (一社)日本糖尿病学会 ) 67 ( Suppl.1 ) S - 251 2024.04

Sampsonの式あるいは人工知能を用いて推定したsmall dense LDLコレステロール値の精度の検証

大久保武志, 遠藤圭佑, 田中希尚, 佐藤達也, 小林亮, 田中真輝人, 秋山幸功, 保坂到, 中田圭, 小山雅之, 大西浩文, 高橋聡, 古橋眞人

糖尿病 ( (一社)日本糖尿病学会 ) 67 ( Suppl.1 ) S - 251 2024.04

Differential Effects of Benzalkonium Chloride on Human Trabecular Meshwork Cells Not Treated or Treated with Transforming Growth Factor-β2 or Dexamethasone.

Megumi Watanabe, Tatsuya Sato, Araya Umetsu, Nami Nishikiori, Megumi Higashide, Masato Furuhashi, Hiroshi Ohguro

Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics 2024.03 [International journal]

Purpose: The objective of the present study was to evaluate the effects of low concentrations of benzalkonium chloride (BAC) (10-7%, 10-6%, or 10-5%) on healthy and glaucomatous human trabecular meshwork (HTM) cells. For this purpose, we used in vitro models replicating a healthy HTM and HTM with primary open-angle glaucoma (POAG) or steroid-induced glaucoma (SG) using two-dimensional (2D) cultures of HTM cells not treated or treated with a 5 ng/mL solution of transforming growth factor-β2 or 250 nM dexamethasone (DEX). Methods: Analyses were carried out for (1) the intercellular affinity function of 2D HTM monolayers, as determined by transepithelial electrical resistance (TEER) measurements; (2) cell viability; (3) cellular metabolism by using a Seahorse bioanalyzer; and (4) expression of extracellular matrix (ECM) molecules, an ECM modulator, and cell junction-related molecules. Results: In the absence and presence of BAC (10-7% or 10-5%), intercellular affinity function determined by TEER and cellular metabolic activities were significantly and dose dependently affected in both healthy and glaucomatous HTM cells despite the fact that there was no significant decrease in cell viabilities. However, the effects based on TEER values were significantly greater in the healthy HTM. The mRNA expression of several molecules that were tested was not substantially modulated by these concentrations of BAC. Conclusions: The findings reported herein suggest that low concentrations of BAC may have unfavorable adverse effects on cellular metabolic capacity by inducing increases in the intercellular affinity properties of the HTM, but those effects of BAC were different in healthy and glaucomatous HTM cells.

Downregulation of Mitochondrial Fusion Protein Expression Affords Protection from Canonical Necroptosis in H9c2 Cardiomyoblasts

Yuki Toda, Sang-Bing Ong, Toshiyuki Yano, Atsushi Kuno, Hidemichi Kouzu, Tatsuya Sato, Wataru Ohwada, Yuki Tatekoshi, Toshifumi Ogawa, Masaki Shimizu, Masaya Tanno, Masato Furuhashi

International Journal of Molecular Sciences ( MDPI AG ) 25 ( 5 ) 2905 - 2905 2024.03

Necroptosis, a form of necrosis, and alterations in mitochondrial dynamics, a coordinated process of mitochondrial fission and fusion, have been implicated in the pathogenesis of cardiovascular diseases. This study aimed to determine the role of mitochondrial morphology in canonical necroptosis induced by a combination of TNFα and zVAD (TNF/zVAD) in H9c2 cells, rat cardiomyoblasts. Time-course analyses of mitochondrial morphology showed that mitochondria were initially shortened after the addition of TNF/zVAD and then their length was restored, and the proportion of cells with elongated mitochondria at 12 h was larger in TNF/zVAD-treated cells than in non-treated cells (16.3 ± 0.9% vs. 8.0 ± 1.2%). The knockdown of dynamin-related protein 1 (Drp1) and fission 1, fission promoters, and treatment with Mdivi-1, a Drp-1 inhibitor, had no effect on TNF/zVAD-induced necroptosis. In contrast, TNF/zVAD-induced necroptosis was attenuated by the knockdown of mitofusin 1/2 (Mfn1/2) and optic atrophy-1 (Opa1), proteins that are indispensable for mitochondrial fusion, and the attenuation of necroptosis was not canceled by treatment with Mdivi-1. The expression of TGFβ-activated kinase (TAK1), a negative regulator of RIP1 activity, was upregulated and the TNF/zVAD-induced RIP1-Ser166 phosphorylation, an index of RIP1 activity, was mitigated by the knockdown of Mfn1/2 or Opa1. Pharmacological TAK1 inhibition attenuated the protection afforded by Mfn1/2 and Opa1 knockdown. In conclusion, the inhibition of mitochondrial fusion increases TAK1 expression, leading to the attenuation of canonical necroptosis through the suppression of RIP1 activity.

Impact of Mitochondrial Dynamics on Necroptosis in Cardiomyocytes(タイトル和訳中)

戸田悠貴, 矢野俊之, 久野篤史, 丹野雅也, 神津英至, 佐藤達也, 大和田渉, 舘越勇輝, 小川俊史, 清水将輝, 古橋眞人

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 88回 PJ080 - 3 2024.03

心筋細胞のネクロプトーシスにおけるミトコンドリア動態の影響(Impact of Mitochondrial Dynamics on Necroptosis in Cardiomyocytes)

戸田悠貴, 矢野俊之, 久野篤史, 丹野雅也, 神津英至, 佐藤達也, 大和田渉, 舘越勇輝, 小川俊史, 清水将輝, 古橋眞人

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 88回 PJ080 - 3 2024.03

ドキソルビシン誘発性心筋症の発症におけるMLKLの寄与(Contribution of MLKL to the Development of Doxorubicin-induced Cardiomyopathy)

清水将輝, 大和田渉, 矢野俊之, 神津英至, 佐藤達也, 長南新太, 小川俊史, 戸田悠貴, 久野篤史, 丹野雅也, 古橋眞人

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 88回 PJ073 - 1 2024.03

Sampsonの式で算出したsmall dense LDLコレステロールは虚血性心疾患発症の有力な予測因子である(Calculated Small Dense Low-density Lipoprotein Cholesterol by Sampson Equation is a Predominant Predictor for the Development of Ischemic Heart Disease)

印鑰雅史, 佐藤達也, 田中希尚, 遠藤圭佑, 保坂到, 三上拓真, 大西浩文, 塙なぎさ, 古橋眞人

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 88回 OJ03 - 2 2024.03

Calculated Small Dense Low-density Lipoprotein Cholesterol by Sampson Equation is a Predominant Predictor for the Development of Ischemic Heart Disease(タイトル和訳中)

印鑰雅史, 佐藤達也, 田中希尚, 遠藤圭佑, 保坂到, 三上拓真, 大西浩文, 塙なぎさ, 古橋眞人

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 88回 OJ03 - 2 2024.03

Contribution of MLKL to the Development of Doxorubicin-induced Cardiomyopathy(タイトル和訳中)

清水将輝, 大和田渉, 矢野俊之, 神津英至, 佐藤達也, 長南新太, 小川俊史, 戸田悠貴, 久野篤史, 丹野雅也, 古橋眞人

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 88回 PJ073 - 1 2024.03

眼圧高値は新規高血圧発症の危険因子である

田中希尚, 佐藤達也, 塙なぎさ, 梅津新矢, 古橋眞人

日本内科学会雑誌 ( (一社)日本内科学会 ) 113 ( 臨増 ) 173 - 173 2024.02

眼圧高値は新規高血圧発症の危険因子である

田中希尚, 佐藤達也, 塙なぎさ, 梅津新矢, 古橋眞人

日本内科学会雑誌 ( (一社)日本内科学会 ) 113 ( 臨増 ) 173 - 173 2024.02

Significant and Various Effects of ML329-Induced MITF Suppression in the Melanoma Cell Line.

Nami Nishikiori, Megumi Watanabe, Tatsuya Sato, Masato Furuhashi, Masae Okura, Tokimasa Hida, Hisashi Uhara, Hiroshi Ohguro

Cancers 16 ( 2 ) 2024.01 [International journal]

To study the inhibitory effects on microphthalmia-associated transcription factor (MITF)-related biological aspects in malignant melanomas (MMs) in the presence or absence of the low-molecular MITF specific inhibitor ML329, cell viability, cellular metabolic functions, and three-dimensional (3D) spheroid formation efficacy were compared among MM cell lines including SK-mel-24, A375, dabrafenib- and trametinib-resistant A375 (A375DT), and WM266-4. Upon exposure to 2 or 10 μM of ML329, cell viability was significantly decreased in WM266-4, SK-mel-24, and A375DT cells, but not A375 cells, in a dose-dependent manner, and these toxic effects of ML329 were most evident in WM266-4 cells. Extracellular flux assays conducted using a Seahorse bioanalyzer revealed that treatment with ML329 increased basal respiration, ATP-linked respiration, proton leakage, and non-mitochondrial respiration in WM266-4 cells and decreased glycolytic function in SK-mel-24 cells, whereas there were no marked effects of ML329 on A375 and A375DT cells. A glycolytic stress assay under conditions of high glucose concentrations also demonstrated that the inhibitory effect of ML329 on the glycolytic function of WM266-4 cells was dose-dependent. In addition, ML329 significantly decreased 3D-spheroid-forming ability, though the effects of ML329 were variable among the MM cell lines. Furthermore, the mRNA expression levels of selected genes, including STAT3 as a possible regulator of 3D spheroid formation, KRAS and SOX2 as oncogenic-signaling-related factors, PCG1a as the main regulator of mitochondrial biogenesis, and HIF1a as a major hypoxia transcriptional regulator, fluctuated among the MM cell lines, possibly supporting the diverse ML329 effects mentioned above. The findings of diverse ML329 effects on various MM cell lines suggest that MITF-associated biological activities are different among various types of MM.

肥満減量術後慢性期の血糖コントロール悪化にセマグルチドが著効した2型糖尿病の症例

赤澤史子, 小川俊史, 中田圭, 佐藤達也, 伊東竜哉, 竹政伊知朗, 古橋眞人

糖尿病 ( (一社)日本糖尿病学会 ) 67 ( 1 ) 37 - 37 2024.01

チルゼパチドが著効した後腹膜滑膜肉腫合併の若年肥満2型糖尿病の症例

大久保武志, 佐藤達也, 小川俊史, 中田圭, 大和田渉, 隅田健太郎, 神津英至, 矢野俊之, 古橋眞人

糖尿病 ( (一社)日本糖尿病学会 ) 67 ( 1 ) 38 - 38 2024.01

中枢性尿崩症合併糖尿病患者に対するSGLT2阻害薬+経口GLP1製剤による治療の1例

中田圭, 佐藤達也, 小川俊史, 大和田渉, 隅田健太郎, 神津英至, 矢野俊之, 古橋眞人

糖尿病 ( (一社)日本糖尿病学会 ) 67 ( 1 ) 40 - 40 2024.01

パッチ式インスリンポンプとリアルタイムCGMを外来で導入し得た症例の検討

吉田十和, 古橋眞人, 佐藤達也

糖尿病 ( (一社)日本糖尿病学会 ) 67 ( 1 ) 41 - 41 2024.01

肥満減量術後慢性期の血糖コントロール悪化にセマグルチドが著効した2型糖尿病の症例

赤澤史子, 小川俊史, 中田圭, 佐藤達也, 伊東竜哉, 竹政伊知朗, 古橋眞人

糖尿病 ( (一社)日本糖尿病学会 ) 67 ( 1 ) 37 - 37 2024.01

中枢性尿崩症合併糖尿病患者に対するSGLT2阻害薬+経口GLP1製剤による治療の1例

中田圭, 佐藤達也, 小川俊史, 大和田渉, 隅田健太郎, 神津英至, 矢野俊之, 古橋眞人

糖尿病 ( (一社)日本糖尿病学会 ) 67 ( 1 ) 40 - 40 2024.01

チルゼパチドが著効した後腹膜滑膜肉腫合併の若年肥満2型糖尿病の症例

大久保武志, 佐藤達也, 小川俊史, 中田圭, 大和田渉, 隅田健太郎, 神津英至, 矢野俊之, 古橋眞人

糖尿病 ( (一社)日本糖尿病学会 ) 67 ( 1 ) 38 - 38 2024.01

Lipid Metabolism Regulators Are the Possible Determinant for Characteristics of Myopic Human Scleral Stroma Fibroblasts (HSSFs).

Hiroshi Ohguro, Araya Umetsu, Tatsuya Sato, Masato Furuhashi, Megumi Watanabe

International journal of molecular sciences 25 ( 1 ) 2023.12 [International journal]

The purpose of the current investigation was to elucidate what kinds of responsible mechanisms induce elongation of the sclera in myopic eyes. To do this, two-dimensional (2D) cultures of human scleral stromal fibroblasts (HSSFs) obtained from eyes with two different axial length (AL) groups, <26 mm (low AL group, n = 2) and >27 mm (high AL group, n = 3), were subjected to (1) measurements of Seahorse mitochondrial and glycolytic indices to evaluate biological aspects and (2) analysis by RNA sequencing. Extracellular flux analysis revealed that metabolic indices related to mitochondrial and glycolytic functions were higher in the low AL group than in the high AL group, suggesting that metabolic activities of HSSF cells are different depending the degree of AL. Based upon RNA sequencing of these low and high AL groups, the bioinformatic analyses using gene ontology (GO) enrichment analysis and ingenuity pathway analysis (IPA) of differentially expressed genes (DEGs) identified that sterol regulatory element-binding transcription factor 2 (SREBF2) is both a possible upstream regulator and a causal network regulator. Furthermore, SREBF1, insulin-induced gene 1 (INSIG1), and insulin-like growth factor 1 (IGF1) were detected as upstream regulators, and protein tyrosine phosphatase receptor type O (PTPRO) was detected as a causal network regulator. Since those possible regulators were all pivotally involved in lipid metabolisms including fatty acid (FA), triglyceride (TG) and cholesterol (Chol) biosynthesis, the findings reported here indicate that FA, TG and Chol biosynthesis regulation may be responsible mechanisms inducing AL elongation via HSSF.

Validation of Estimated Small Dense Low-Density Lipoprotein Cholesterol Concentration in a Japanese General Population.

Keisuke Endo, Ryo Kobayashi, Makito Tanaka, Marenao Tanaka, Yukinori Akiyama, Tatsuya Sato, Itaru Hosaka, Kei Nakata, Masayuki Koyama, Hirofumi Ohnishi, Satoshi Takahashi, Masato Furuhashi

Journal of atherosclerosis and thrombosis 2023.12 [Domestic journal]

AIM: A high level of directly measured small dense low-density lipoprotein cholesterol (sdLDL-C) is a strong risk factor for atherosclerotic cardiovascular disease. A method for estimating sdLDL-C by using Sampson's equation that includes levels of total cholesterol, high-density lipoprotein cholesterol (HDL-C), non-HDL-C and triglycerides (TG) has recently been proposed. We investigated the validation and exploration of estimated sdLDL-C level. METHODS: The associations between measured and estimated sdLDL-C levels were investigated in 605 Japanese subjects (men/women: 280/325; mean age: 65±15 years) who received annual health check-ups in the Tanno Sobetsu Study, a population-based cohort. RESULTS: Estimated sdLDL-C level was highly correlated with measured sdLDL-C level in all subjects (R2 =0.701), nondiabetic subjects without any medication (n=254, R 2=0.686) and subjects with diabetes mellitus (n=128, R2=0.721). Multivariable regression analysis showed that levels of non-HDL-C, TG and γ-glutamyl transpeptidase (γGTP) were independent predictors of measured sdLDL-C level. In a stratification of the LDL window, all of the subjects with a combination of high non-HDL-C (≥ 170 mg/dL) and high TG (≥ 150 mg/dL) had high levels of measured and estimated sdLDL-C (≥ 35 mg/dL). Furthermore, machine learning-based estimation of sdLDL-C level by artificial intelligence software, Prediction One, was substantially improved by using components of Sampson's equation (R2=0.803) and by using those components with the addition of γGTP and deletion of TC (R2=0.929). CONCLUSIONS: sdLDL-C level estimated by Sampson's equation can be used instead of measured sdLDL-C level in general practice. By building multiple machine learning models of artificial intelligence, a more accurate and practical estimation of sdLDL-C level might be possible.

Benzalkonium chloride greatly deteriorates the biological activities of human corneal stroma fibroblasts in a concentration-dependent manner.

Araya Umetsu, Yosuke Ida, Tatsuya Sato, Masato Furuhashi, Hiroshi Ohguro, Megumi Watanabe

Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie 2023.12 [International journal]

BACKGROUND: Corneal tissues indirectly obtain nutritional needs and oxygen to maintain their homeostasis, and therefore, benzalkonium chloride (BAC) containing ocular instillations for medical therapy may, in turn, induce toxic effects more than expected in corneal tissues, especially the inside stroma layer. METHODS: To evaluate the effects of very low concentrations (10-8%, 10-6%, or 10-4%) of BAC on human corneal stroma, we used two-dimensional (2D) cultures of human corneal stromal fibroblast (HCSF) cells and carried out the following analyses: (1) cell viability measurements, (2) Seahorse cellular bio-metabolism analysis, and (3) the expression of ECM molecules and endoplasmic reticulum (ER) stress-related molecules. RESULTS: In the absence and presence of 10-8%, 10-6%, or 10-4% concentrations of BAC, cell viability deteriorated and this deterioration was dose-dependent. The results showed that maximal mitochondrial respiration was decreased, the mRNA expression of most of ECM proteins was decreased, and ER stress-related molecules were substantially and dose-dependently down-regulated in HCSFs by the BAC treatment. CONCLUSIONS: The findings reported herein indicate that the presence of BAC, even at such low concentrations, is capable of causing the deterioration of cellular metabolic functions and negatively affecting the response to ER stress in HCSF cells resulting in a substantially decreased cellular viability.

Coexistence of sarcopenia and self‐reported weight loss is a powerful predictor of mortality in older patients with heart failure

Katsuhiko Ohori, Toshiyuki Yano, Satoshi Katano, Ryohei Nagaoka, Ryo Numazawa, Kotaro Yamano, Yusuke Fujisawa, Hidemichi Kouzu, Nobutaka Nagano, Takefumi Fujito, Ryo Nishikawa, Wataru Ohwada, Masato Furuhashi

Geriatrics & Gerontology International ( Wiley ) 2023.12

Aim We examined whether the addition of self‐reported weight loss improves the accuracy of prediction of mortality caused by sarcopenia in heart failure (HF) patients. Methods We enrolled 477 HF patients (mean age 77 years) who received combined assessment of sarcopenia and self‐reported weight loss. Sarcopenia was diagnosed according to the criteria of the Asian Working Group for Sarcopenia. If the patients answered “yes” to the question “have you lost 2 kg or more in the past 6 months?”, they were diagnosed as having self‐reported weight loss. Results Sarcopenia and self‐reported weight loss coexisted in 32% of patients. During a median follow‐up period of 763 days, 65 patients (15%) died. Kaplan–Meier curves showed a significantly higher rate of mortality in HF patients with both sarcopenia and self‐reported weight loss than in HF patients with sarcopenia alone. Multivariate Cox proportional hazards analysis showed that the coexistence of sarcopenia and self‐reported weight loss is an independent predictor of mortality in HF patients. Inclusion of the coexistence of sarcopenia and self‐reported weight loss in the baseline model consisting of established prognostic markers significantly improved both the net reclassification index and the integrated discrimination index. Conclusions The coexistence of sarcopenia and self‐reported weight loss is a powerful predictor of mortality in HF patients. Geriatr Gerontol Int 2023; ••: ••–••.

Systemic sarcoidosis presenting as a rare combination of interstitial nephritis with necrotizing vasculitis and urinary retention due to prostate involvement: a case report.

Arata Osanami, Tomohisa Yamashita, Shintaro Sakurada, Tatsuya Sato, Yuki Kyoda, Tetsuya Shindo, Hiromi Fujita, Yayoi Ogawa, Masato Furuhashi

BMC nephrology 24 ( 1 ) 370 - 370 2023.12 [International journal]

BACKGROUND: Sarcoidosis affects multiple organs and exhibits diverse clinical manifestations. Although tubulointerstitial nephritis is a known feature of renal involvement, necrotizing vasculitis is rare. Furthermore, prostate involvement with urinary retention is unusual in patients with sarcoidosis. Here, we report a case of systemic sarcoidosis with a rare combination of manifestations and different acute kidney injuries. CASE PRESENTATION: A 66-year-old man developed sudden urinary retention and fever. He was diagnosed with prostatitis and admitted to our hospital. An indwelling urethral catheter was inserted, and antimicrobial therapy was initiated; however, the prostatitis was refractory. Computed tomography revealed enlarged mediastinal lymph nodes. Analysis of transbronchoscopic lymph node and prostate biopsies showed epithelioid cell granulomas, suggesting systemic sarcoidosis. During the clinical course, the serum creatinine level rapidly increased to 2.36 mg/dL without oliguria. A kidney biopsy revealed tubulointerstitial injury with moderate lymphohistiocytic infiltration and small-vessel vasculitis in the interstitium. Following oral administration of 60 mg/day prednisolone, the patient's renal function immediately improved, and urinary retention did not recur. CONCLUSIONS: To the best of our knowledge, this is the first reported case of sarcoidosis with two unusual complications. Given its clinical course and pathology, this case is clinically valuable.

Relationship between serum iron level and physical function in heart failure patients is lost by presence of diabetes.

Katsuhiko Ohori, Toshiyuki Yano, Satoshi Katano, Ryohei Nagaoka, Ryo Numazawa, Kotaro Yamano, Yusuke Fujisawa, Hidemichi Kouzu, Nobutaka Nagano, Takefumi Fujito, Ryo Nishikawa, Wataru Ohwada, Tatsuya Sato, Masato Furuhashi

ESC heart failure 2023.12 [International journal]

AIMS: Iron deficiency (ID) is common in patients with heart failure (HF) and is reportedly associated with exercise intolerance and impaired quality of life. Iron supplementation therapy in HF patients with ID improves exercise capacity. Conversely, protective roles of iron depletion in the development of diabetes mellitus (DM) and its complications have been proposed. This study aimed to determine the impact of ID on physical function in HF patients with and without DM. METHODS AND RESULTS: We enrolled consecutive patients who were admitted to our institute for HF diagnosis and management. The short physical performance battery (SPPB) was used to evaluate physical function, and low physical function was defined as an SPPB score of <10 points as individuals with SPPB scores of <10 points are most likely to be classified as frail and are at high risk for disability and future adverse events, including death. ID was defined as serum ferritin < 100 or 100-299 ng/mL when transferrin saturation (TSAT) was <20% according to the HF guidelines. Among the 562 HF patients (72 ± 14 years old; 56% male), 329 patients (58%) and 191 patients (34%) had ID and low physical function, respectively. Multivariate logistic regression analysis showed that TSAT as a continuous variable, but not ID, was a predictor of low physical function (odds ratio: 0.980, P = 0.024). Subgroup analysis showed that a significant association between low TSAT and low physical function was lost in HF patients with DM (P for interaction < 0.001). A spline dose-response curve for the relationship between TSAT and risk of low physical function with adjustments for covariates associated with low physical function in non-DM patients was almost linear with an increase in the risk of low physical function as the TSAT increased, but such a relationship was not found in the analyses of DM patients. A lack of close TSAT-SPPB relationship in HF patients with DM was confirmed also in a propensity-score-matched cohort. CONCLUSIONS: TSAT as a continuous variable, but not ID, was independently associated with physical function in HF patients, and a significant association was lost in patients with HF and DM, suggesting a limited impact of iron supplementation therapy in HF patients with DM.

Predictive modeling for the development of diabetes mellitus using key factors in various machine learning approaches

Marenao Tanaka, Yukinori Akiyama, Kazuma Mori, Itaru Hosaka, Kenichi Kato, Keisuke Endo, Toshifumi Ogawa, Tatsuya Sato, Toru Suzuki, Toshiyuki Yano, Hirofumi Ohnishi, Nagisa Hanawa, Masato Furuhashi

Diabetes Epidemiology and Management ( Elsevier BV ) 100191 - 100191 2023.12

Circulating tumor necrosis factor-related biomarkers predict kidney function decline in Japanese patients with diabetes: An observational cohort study.

Maki Murakoshi, Nozomu Kamei, Yusuke Suzuki, Mitsunobu Kubota, Michiyoshi Sanuki, Hirotaka Tashiro, Takumi Iwasawa, Kazunori Kato, Marenao Tanaka, Masato Furuhashi, Tomohito Gohda

Diabetes research and clinical practice 206 111017 - 111017 2023.11 [International journal]

AIMS: Tumor necrosis factor (TNF) receptors (TNFRs: TNFR1 and, TNFR2) are reportedly associated with chronic kidney disease (CKD) progression chiefly in Caucasian patients with diabetes. We assessed the prognostic value of TNF-related biomarkers for CKD progression in Japanese patients with diabetes. METHODS: We estimated TNF-related biomarkers using an enzyme-linked immunosorbent assay in 640 patients with diabetes. Cox proportional hazards analysis was performed to estimate hazard ratios (HRs) per one standard deviation (SD) increase in a log-transformed biomarker. The kidney and the composite outcome were defined as a 30% reduction in estimated glomerular filtration rate (eGFR) from baseline, and kidney outcome plus death before kidney outcome, respectively. RESULTS: During the median follow-up of 5.4 years, 75 (11.7%) patients reached the kidney outcome and 37 (5.8%) died before reaching the kidney outcome. Each SD increase in baseline circulating TNFR1, TNFR2, and ephrin type-A receptor 2 (EphA2) was associated with a higher risk of the kidney outcome independently from baseline eGFR and urine albumin-to-creatinine ratio. However, circulating osteoprotegerin was associated with the composite outcome only. CONCLUSIONS: Elevated TNFR1, TNFR2, and EphA2 were associated with both kidney and composite outcomes in Japanese patients with diabetes.

A high level of thyroid-stimulating hormone is a risk factor for the development of chronic kidney disease in men: a 10-year cohort study.

Keisuke Endo, Marenao Tanaka, Tatsuya Sato, Kazuma Mori, Itaru Hosaka, Takuma Mikami, Araya Umetsu, Yukinori Akiyama, Hirofumi Ohnishi, Nagisa Hanawa, Masato Furuhashi

Hypertension research : official journal of the Japanese Society of Hypertension 2023.10 [International journal]

Hypothyroidism has been reported to be associated with chronic kidney disease (CKD). However, the impact of thyroid-stimulating hormone (TSH) on new onset of CKD and its gender dependence remain undetermined. We investigated the association of serum TSH level and the development of CKD defined by estimated glomerular filtration rate (eGFR) < 60 mL/min/1.73 m2 or positive for urine protein in 28,990 Japanese subjects who received annual health examinations. After excluding subjects with no data for serum TSH, urinalysis and eGFR and those with CKD at baseline, a total of 10,392 subjects (men/women: 6802/3590, mean age: 48 years) were recruited. During a 10-year follow-up, 1185 men (6.7%) and 578 women (2.9%) newly developed CKD. Multivariable Cox proportional hazard models after adjustment of age, body mass index, smoking habit, hypertension, diabetes mellitus, dyslipidemia, ischemic heart disease and eGFR (≥ 90 mL/min/1.73 m2) showed that the hazard ratio (HR) for the development of CKD in the high TSH (> 4.2 mU/L) group was significantly higher than that in the low TSH (≤ 4.2 mU/L) group in men (HR [95% confidence interval]: 1.41 [1.09-1.83]) but not in women (1.08 [0.77-1.51]). There was a significant interaction between sex and the category of TSH level for the development of CKD (p = 0.02). The adjusted HR with a restricted cubic spline increased with a higher level of TSH in men but not in women. In conclusion, a high level of TSH is associated with an increased risk for the development of CKD in men but not in women.

Independent Link Between Use of Mineralocorticoid Receptor Antagonists and Muscle Wasting in Heart Failure Patients Not Receiving Renin-Angiotensin System Inhibitors

Ryo Numazawa, Satoshi Katano, Toshiyuki Yano, Ryohei Nagaoka, Katsuhiko Ohori, Hidemichi Kouzu, Suguru Honma, Yusuke Fujisawa, Kotaro Yamano, Arata Osanami, Masayuki Koyama, Akiyoshi Hashimoto, Masato Furuhashi

Circulation Journal ( Japanese Circulation Society ) 2023.10

3D culture induction of adipogenic differentiation in 3T3-L1 preadipocytes exhibits adipocyte-specific molecular expression patterns and metabolic functions

Keisuke Endo, Tatsuya Sato, Araya Umetsu, Megumi Watanabe, Fumihito Hikage, Yosuke Ida, Hiroshi Ohguro, Masato Furuhashi

Heliyon ( Elsevier BV ) 9 ( 10 ) e20713 - e20713 2023.10

HIF-PH阻害薬の中止因子の検討

宮尾舞, 石郷友之, 相神智宏, 野々山雅俊, 山下智久, 小山雅之, 藤居賢, 矢野俊之, 古橋眞人, 福土将秀

日本腎臓病薬物療法学会誌 ( 日本腎臓病薬物療法学会 ) 12 ( 特別号 ) S200 - S200 2023.10

非透析症例におけるHIF-PH阻害薬のeGFR保持効果の検討

相神智宏, 石郷友之, 宮尾舞, 野々山雅俊, 山下智久, 小山雅之, 藤居賢, 矢野俊之, 古橋眞人, 福土将秀

日本腎臓病薬物療法学会誌 ( 日本腎臓病薬物療法学会 ) 12 ( 特別号 ) S200 - S200 2023.10

Optimal serum 25-hydroxyvitamin D level to prevent sarcopenia in patients with heart failure: insights from a dose-response relationship

Ryohei Nagaoka, Satoshi Katano, Toshiyuki Yano, Ryo Numazawa, Kotaro Yamano, Yusuke Fujisawa, Suguru Honma, Tatsuki Kamoda, Kohei Sato, Hidemichi Kouzu, Katsuhiko Ohori, Masaki Katayose, Akiyoshi Hashimoto, Masato Furuhashi

Nutrition, Metabolism and Cardiovascular Diseases ( Elsevier BV ) 2023.10

心臓血管外科における未来医療を目指して:予測医学・シミュレーションを用いたPrecision medicine Random survival forest法を用いた機械学習による術後予後予測の有用性に関する検討生存時間解析における古典的Cox比例ハザードモデルとの比較と課題

保坂到, 田中希尚, 秋山幸功, 佐藤亮太, 森和真, 小川俊史, 梅津新矢, 遠藤圭佑, 三上拓真, 佐藤達也, 梅田璃子, 宇塚武司, 古橋眞人, 川原田修義

日本胸部外科学会定期学術集会 ( (一社)日本胸部外科学会 ) 76回 CSY3 - 4 2023.10

心臓血管外科における未来医療を目指して:予測医学・シミュレーションを用いたPrecision medicine Random survival forest法を用いた機械学習による術後予後予測の有用性に関する検討生存時間解析における古典的Cox比例ハザードモデルとの比較と課題

保坂到, 田中希尚, 秋山幸功, 佐藤亮太, 森和真, 小川俊史, 梅津新矢, 遠藤圭佑, 三上拓真, 佐藤達也, 梅田璃子, 宇塚武司, 古橋眞人, 川原田修義

日本胸部外科学会定期学術集会 ( (一社)日本胸部外科学会 ) 76回 CSY3 - 4 2023.10

TGF-β2 Induces Epithelial-Mesenchymal Transitions in 2D Planer and 3D Spheroids of the Human Corneal Stroma Fibroblasts in Different Manners.

Araya Umetsu, Yosuke Ida, Tatsuya Sato, Masato Furuhashi, Hiroshi Ohguro, Megumi Watanabe

Biomedicines 11 ( 9 ) 2023.09 [International journal]

To examine the epithelial-mesenchymal transition (EMT) that is induced on the human corneal stroma, two- and three-dimensional (2D and 3D) cultures of human corneal stroma fibroblasts (HCSFs) were used. In this study, HCSF 2D monolayers and 3D spheroids were characterized by (1) scanning electron microscopy (SEM), (2) trans-endothelial electrical resistance (TEER) measurements and fluorescein isothiocyanate (FITC)-dextran permeability, (3) cellular metabolic measurements, (4) the physical properties of 3D HCSF spheroids, and (5) the extracellular matrix (ECM) molecule gene expressions, including collagen (COL) 1, 4 and 6, and fibronectin (FN), a tissue inhibitor of metalloproteinase (TIMP) 1-4, matrix metalloproteinase (MMP) 2, 3, 9 and 14, and several endoplasmic reticulum (ER) stress-related factors. In the 2D HCSFs, TGF-β2 concentration-dependently generated (1) a considerable increase in ECM deposits revealed by SEM, (2) an increase in TEER values and a decrease in FITC-dextran permeability, (3) increases in both mitochondrial and glycolytic functions, and a substantial upregulation of COL1, COL4, FN, αSMA, TIMP1, TIMP, and most ER stress-related genes and the downregulation of COL6 and MMP3. In the case of 3D spheroids, TGF-β2 induced the downsizing and stiffening of 3D spheroids and the upregulation of COL6, MMP14, and most ER stress-related genes. These findings suggest that TGF-β2 significantly induced a number of EMT-associated biological events including planar proliferation, cellular metabolic functions, and the production of ECM molecules in the 2D cultured HCSF cells, but these effects were significantly less pronounced in the case of 3D HCSF spheroids.

Circulating level of β-aminoisobutyric acid (BAIBA), a novel myokine-like molecule, is inversely associated with fat mass in patients with heart failure

Satoshi Katano, Toshiyuki Yano, Hidemichi Kouzu, Ryohei Nagaoka, Ryo Numazawa, Kotaro Yamano, Yusuke Fujisawa, Katsuhiko Ohori, Nobutaka Nagano, Takefumi Fujito, Ryo Nishikawa, Wataru Ohwada, Masaki Katayose, Tatsuya Sato, Atsushi Kuno, Masato Furuhashi

Heart and Vessels ( Springer Science and Business Media LLC ) 2023.09

Fatty Liver Indexを用いて高血圧発症を予測する機械学習モデルの構築

田中希尚, 秋山幸功, 佐藤亮太, 保坂到, 三上拓真, 遠藤圭佑, 梅津新矢, 小川俊史, 森和真, 佐藤達也, 塙なぎさ, 古橋眞人

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 45回 391 - 391 2023.09

心不全患者におけるレニン-アンジオテンシン系活性化は骨格筋量の低下と関連する

沼澤瞭, 片野唆敏, 矢野俊之, 長岡凌平, 神津英至, 大堀克彦, 藤澤佑輔, 山埜光太郎, 長南新太, 小山雅之, 橋本暁佳, 古橋眞人

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 45回 398 - 398 2023.09

Calculated Small Dense Low-Density Lipoprotein Cholesterol Level is A Predominant Predictor for New Onset of Ischemic Heart Disease.

Masafumi Inyaku, Marenao Tanaka, Tatsuya Sato, Keisuke Endo, Kazuma Mori, Itaru Hosaka, Takuma Mikami, Araya Umetsu, Hirofumi Ohnishi, Yukinori Akiyama, Nagisa Hanawa, Masato Furuhashi

Journal of atherosclerosis and thrombosis 2023.08 [Domestic journal]

AIM: A high level of directly measured small dense low-density lipoprotein cholesterol (sdLDL-C) is a strong risk factor for ischemic heart disease (IHD). However, it remains unclear whether estimated sdLDL-C level is a predictor for IHD. We investigated the associations of new onset of IHD with levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), non-HDL-C, triglycerides (TG), LDL-C and calculated sdLDL-C by Sampson's equation. METHODS: After exclusion of subjects with IHD or those with TG ≥ 800 mg/dL, a total of 18,176 subjects (men/women: 11,712/6,464, mean age: 46 years) were recruited among 28,990 Japanese individuals who received annual health checkups. RESULTS: During the 10-year follow-up period, 456 men (3.9%) and 121 women (1.9%) newly developed IHD. Multivariable Cox proportional hazard analyses after adjustment of age, sex, obesity, smoking habit, family history of IHD, estimated glomerular filtration rate, hypertension and diabetes mellitus at baseline showed that the hazard ratio (HR) (1.38 [95% confidence interval: 1.03-1.85]) for new onset of IHD in subjects with the 4th quartile (Q4) of sdLDL-C (≥ 42 mg/dL) was significantly higher than that in subjects with the 1st quartile (Q1) (≤ 24 mg/dL) as the reference, though the adjusted HRs in subjects with Q2-Q4 of TC, HDL-C, non-HDL-C, LDL-C and TG were comparable with those in subjects with Q1 of the respective lipid fractions. The adjusted HR with a restricted cubic spline increased with a higher level of calculated sdLDL-C as a continuous value at baseline. CONCLUSIONS: sdLDL-C level calculated by Sampson's equation is a predominant predictor for the development of IHD in a general Japanese population.

Physical Properties and Cellular Metabolic Characteristics of 3D Spheroids Are Possible Definitive Indices for the Biological Nature of Cancer-Associated Fibroblasts.

Nami Nishikiori, Kohichi Takada, Tatsuya Sato, Sho Miyamoto, Megumi Watanabe, Yui Hirakawa, Shohei Sekiguchi, Masato Furuhashi, Akira Yorozu, Kenichi Takano, Akihiro Miyazaki, Hiromu Suzuki, Hiroshi Ohguro

Cells 12 ( 17 ) 2023.08 [International journal]

The current study's objective was to elucidate some currently unknown biological indicators to evaluate the biological nature of cancer-associated fibroblasts (CAFs). For this purpose, four different CAFs, CAFS1, CAFS2, SCC17F and MO-1000, were established using surgical specimens from oral squamous cell carcinomas (OSCC) with different clinical malignant stages (CAFS1 and CAFS2, T2N0M0, stage II; SCC17F and MO-1000, T4aN2bM0, stage IVA). Fibroblasts unrelated to cancer (non-CAFs) were also prepared and used as controls. Initially, confirmation that these four fibroblasts were indeed CAFs was obtained by their mRNA expression using positive and negative markers for the CAF or fibroblasts. To elucidate possible unknown biological indicators, these fibroblasts were subjected to a cellular metabolic analysis by a Seahorse bioanalyzer, in conjugation with 3D spheroid cultures of the cells and co-cultures with a pancreas ductal carcinoma cell line, MIA PaCa-2. The mitochondrial and glycolytic functions of human orbital fibroblasts (HOF) were nearly identical to those of Graves'-disease-related HOF (GOF). In contrast, the characteristics of the metabolic functions of these four CAFs were different from those of human conjunctival fibroblasts (HconF), a representative non-CAF. It is particularly noteworthy that CAFS1 and CAFS2 showed markedly reduced ratios for the rate of oxygen consumption to the extracellular acidification rate, suggesting that glycolysis was enhanced compared to mitochondrial respiration. Similarly, the physical aspects, their appearance and stiffness, of their 3D spheroids and fibroblasts that were induced effects based on the cellular metabolic functions of MIA PaCa-2 were also different between CAFs and non-CAFs, and their levels for CAFS1 or SCC17F were similar to those for CAFS2 or MO-1000 cells, respectively. The findings reported herein indicate that cellular metabolic functions and the physical characteristics of these types of 3D spheroids may be valuable and useful indicators for estimating potential biological diversity among various CAFs.

Development and validation of osteoporosis risk assessment score, ORAS, in heart failure patients: comparison with the Osteoporosis Self-Assessment Tool for Asians (OSTA)

Ryo Numazawa, Satoshi Katano, Toshiyuki Yano, Masayuki Koyama, Ryohei Nagaoka, Yusuke Fujisawa, Kotaro Yamano, Suguru Honma, Katsuhiko Ohori, Hidemichi Kouzu, Masaki Katayose, Masato Furuhashi, Kazufumi Tsuchihashi, Akiyoshi Hashimoto

European Journal of Cardiovascular Nursing ( Oxford University Press (OUP) ) 2023.08

Abstract Aim Osteoporosis is prevalent and is associated with poor prognosis in heart failure (HF) patients. However, bone mineral density (BMD) measurement by a dual-energy X-ray absorptiometry (DEXA) scan is not always available in a daily clinical setting and large-scale population-based studies. Methods A single-center, cross-sectional observational study was conducted with 387 patients (median age: 77 years [interquartile range: 68 to 83 years]; 37% women). BMDs were measured by DEXA scans, and osteoporosis was diagnosed as ≤ -2.5 standard deviation of the BMDs in healthy young adults. Osteoporosis risk assessment score (ORAS) was developed using significant predictors from a logistic regression model for osteoporosis and was subsequently validated. Results Osteoporosis was found in 103 (27%) of the 387 HF patients. Multivariate logistic regression analyses yielded the ORAS based on sex, BMI, handgrip strength, and anti-coagulant therapy utilization. The C-index of ORAS in the developmental set (0.796, 95% confidence interval: 0.747 to 0.845) was similar to the bootstrap validation of the prediction model (0.784), and tended to be higher than that of the Osteoporosis Self-Assessment Tool for Asians (OSTA). A nomogram of ORAS, established on the basis of the final logistic regression model, demonstrated 100% sensitivity at the lowest score (35 points), with an optimal cutoff point of 127 points, yielding 85% sensitivity and 62% specificity. Conclusions ORAS exhibits superior predictive performance to OSTA in predicting osteoporosis in HF patients, establishing itself as a valuable tool for early detection in both daily clinical practice and large-scale population-based studies.

An increase in calculated small dense low-density lipoprotein cholesterol predicts new onset of hypertension in a Japanese cohort.

Marenao Tanaka, Tatsuya Sato, Keisuke Endo, Masafumi Inyaku, Kazuma Mori, Itaru Hosaka, Takuma Mikami, Yukinori Akiyama, Hirofumi Ohnishi, Nagisa Hanawa, Masato Furuhashi

Hypertension research : official journal of the Japanese Society of Hypertension 2023.08 [International journal]

A disorder of lipid metabolism is involved in cardiovascular diseases including hypertension. A high level of small dense low-density lipoprotein cholesterol (sdLDL-C) is a strong risk factor for atherosclerotic cardiovascular disease. However, the association between sdLDL-C and hypertension has not been fully investigated. We investigated the associations between the development of hypertension during a 10-year period and levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), non-HDL-C, triglycerides (TG), and LDL-C and sdLDL-C calculated by using the Sampson equations in 28,990 Japanese subjects who received annual health examinations. After exclusion of subjects with missing data, those with hypertension, and those with TG ≥ 800 mg/dL at baseline, a total of 15,177 subjects (men/women: 9374/5803, mean age: 46 years) were recruited. During the 10-year period, 2379 men (25.4%) and 724 women (12.5%) had new onset of hypertension. Multivariable Cox proportional hazard model analyses showed that levels of HDL-C, non-HDL-C, TG and sdLDL-C, but not levels of TC and LDL-C, were independent risk factors for the development of hypertension after adjustment of age, sex, family history of hypertension, systolic blood pressure, obesity, current smoking habit, alcohol drinking habit, estimated glomerular filtration rate, diagnosis of diabetes mellitus and use of lipid-lowering drugs and that the adjusted risk of sdLDL-C (per 1-standard deviation) was highest (hazard ratio [95% confidence interval: 1.09 [1.05-1.13]). The addition of sdLDL-C to traditional risk factors for hypertension significantly improved the discriminatory capability, which was better than that of other lipid fractions. In conclusion, a high level of calculated sdLDL-C predicts the development of hypertension.

Coexistence of Metabolic Dysfunction-Associated Fatty Liver Disease and Chronic Kidney Disease Is a More Potent Risk Factor for Ischemic Heart Disease.

Daisuke Miyamori, Marenao Tanaka, Tatsuya Sato, Keisuke Endo, Kazuma Mori, Takuma Mikami, Itaru Hosaka, Nagisa Hanawa, Hirofumi Ohnishi, Masato Furuhashi

Journal of the American Heart Association 12 ( 14 ) e030269 2023.07 [International journal]

Background Metabolic dysfunction-associated fatty liver disease (MAFLD), defined as fatty liver with overweight/obesity, type 2 diabetes, or metabolic abnormalities, is a newly proposed disease. However, it remains unclear whether the coexistence of MAFLD and chronic kidney disease (CKD) is a more potent risk factor for ischemic heart disease (IHD). Methods and Results We investigated the risk of the combination of MAFLD and CKD for development of IHD during a 10-year follow-up period in 28 990 Japanese subjects who received annual health examinations. After exclusion of subjects without data for abdominal ultrasonography or with the presence of IHD at baseline, a total of 14 141 subjects (men/women: 9195/4946; mean age, 48 years) were recruited. During the 10-year period (mean, 6.9 years), 479 subjects (men/women, 397/82) had new onset of IHD. Kaplan-Meier survival curves showed significant differences in rates of the cumulative incidence of IHD in subjects with and those without MAFLD (n=4581) and CKD (n=990; stages 1/2/3/4-5, 198/398/375/19). Multivariable Cox proportional hazard model analyses showed that coexistence of MAFLD and CKD, but not MAFLD or CKD alone, was an independent predictor for development of IHD after adjustment for age, sex, current smoking habit, family history of IHD, overweight/obesity, diabetes, hypertension, and dyslipidemia (hazard ratio, 1.51 [95% CI, 1.02-2.22]). The addition of the combination of MAFLD and CKD to traditional risk factors for IHD significantly improved the discriminatory capability. Conclusions The coexistence of MAFLD and CKD predicts new onset of IHD better than does MAFLD or CKD alone.

Prostanoid FP and EP2 Receptor Agonists Induce Epithelial and Subepithelial Fibrogenetic Changes in Human Conjunctival Fibroblasts in Different Manners.

Yuri Tsugeno, Tatsuya Sato, Megumi Watanabe, Masato Furuhashi, Hiroshi Ohguro

Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics 39 ( 6 ) 404 - 414 2023.07 [International journal]

Purpose: To examine the effects of prostanoid FP and EP2 receptor agonists, PGF2α and Omidenepag (OMD), respectively, on the transforming growth factor beta (TGF-β2) induced conjunctival fibrogenesis. Methods: Two-dimension (2D) and three-dimension (3D) cultures of these fibroblasts were subjected to following analyses: (1) planar proliferation evaluated by transendothelial electron resistance (TEER) measurements, (2) real-time metabolic analyses, (3) subepithelial proliferation evaluated by 3D spheroid' size and stiffness measurements, and (4) the mRNA expression of extracellular matrix (ECM) molecules and their modulators. Results: TGF-β2 induced increase in the planar proliferation was significantly decreased or enhanced by PGF2α or OMD, respectively. The proportion of oxygen consumption required to drive ATP synthesis compared with that driving proton leakage was increased by PGF2α and OMD independently with TGF-β2. In contrast, maximal mitochondrial respiration was decreased by PGF2α and OMD, and the OMD-induced effect was further enhanced by the presence of TGF-β2. In addition, the TGF-β2 dependent increase in the glycolytic capacity was cancelled by PGF2α and/or OMD. Alternatively, subepithelial proliferation, as evidenced by the stiffness of the 3D spheroids, was substantially increased by both PGF2α and OMD, and these were differently modulated by TGF-β2. The expression of several related factors as above fluctuated among the conditions for both 2D and 3D and TGF-β2 untreated or treated cultures. Conclusion: The present findings indicate that the prostanoid FP or the EP2 receptor agonist may solely and differently induce the planar and subepithelial proliferation of HconF cells and these were also modulated by TGF-β2.

TGF-β Isoforms Affect the Planar and Subepithelial Fibrogenesis of Human Conjunctival Fibroblasts in Different Manners

Megumi Watanabe, Yuri Tsugeno, Tatsuya Sato, Araya Umetsu, Nami Nishikiori, Masato Furuhashi, Hiroshi Ohguro

Biomedicines 2023.07 [Refereed]

mTOR Inhibitors Modulate the Physical Properties of 3D Spheroids Derived from H9c2 Cells

Megumi Watanabe, Toshiyuki Yano, Tatsuya Sato, Araya Umetsu, Megumi Higashide, Masato Furuhashi, Hiroshi Ohguro

International Journal of Molecular Sciences 2023.07 [Refereed]

Simultaneous Effects of a Selective EP2 Agonist, Omidenepag, and a Rho-Associated Coiled-Coil Containing Protein Kinase Inhibitor, Ripasudil, on Human Orbital Fibroblasts.

Fumihito Hikage, Megumi Watanabe, Tatsuya Sato, Araya Umetsu, Yuri Tsugeno, Masato Furuhashi, Hiroshi Ohguro

Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics 2023.06 [International journal]

Purpose: To assess the combined effects of omidenepag (OMD), a selective EP2 agonist, and ripasudil (Rip), an inhibitor of rho-associated coiled-coil containing protein kinases, on the human orbital adipose tissue, two-dimensional (2D) or three-dimensional (3D) cultures of human orbital fibroblasts (HOFs) were employed. Methods: Cellular metabolic functions (2D), physical (3D), lipid staining (3D), and quantitative polymerase chain reaction for adipogenesis-related genes, PPARγ and AP2, and extracellular matrix (ECM) molecules, including collagen (COL)1, 4, and 6, and fibronectin (FN) (3D) were evaluated in the presence of OMD (100 nM) and/or Rip (10 μM). Results: Real-time metabolic analyses revealed that the adipogenic differentiation (DIF+) with OMD significantly shifted an energetic state toward energetic, whereas DIF+ with Rip significantly shifted that toward quiescent. In the case of both drugs upon DIF+, the metabolic effect of OMD was predominant. DIF+ induced enlargement and stiffed 3D spheroid with increased lipid staining and mRNA expression of adipogenesis-related genes, COL4 and COL6, and decreased the expression of COL1. In the presence of OMD and/or Rip to DIF+, (1) the sizes were further increased by Rip and the stiffness was significantly decreased by OMD or Rip and (2) COL4 or AP2 expression was substantially increased by OMD or Rip, respectively. Conclusion: The results presented herein indicate that the metabolic effects of OMD and Rip exerted opposing effects and the effects of OMD toward Ap2 and ECM expressions were distinct from those of Rip, but the effects of OMD toward the physical aspects and adipogenesis of the 3D cultured HOFs were similar to the effects of Rip.

Impact of Radar Chart-Based Information Sharing in a Multidisciplinary Team on In-Hospital Outcomes and Prognosis in Older Patients With Heart Failure

Satoshi Katano, Toshiyuki Yano, Ryo Numazawa, Ryohei Nagaoka, Kotaro Yamano, Yusuke Fujisawa, Suguru Honma, Ayako Watanabe, Katsuhiko Ohori, Hidemichi Kouzu, Takefumi Fujito, Tomoyuki Ishigo, Hayato Kunihara, Hiroya Fujisaki, Masaki Katayose, Akiyoshi Hashimoto, Masato Furuhashi

Circulation Reports ( Japanese Circulation Society ) 2023.06

Independent Association Between Use of Proton Pump Inhibitors and Muscle Wasting in Patients with Heart Failure: A Single-Center, Ambispective, Observational Study

Katsuhiko Ohori, Toshiyuki Yano, Satoshi Katano, Ryohei Nagaoka, Ryo Numazawa, Kotaro Yamano, Yusuke Fujisawa, Hidemichi Kouzu, Masayuki Koyama, Nobutaka Nagano, Takefumi Fujito, Ryo Nishikawa, Wataru Ohwada, Masato Furuhashi

Drugs & Aging ( Springer Science and Business Media LLC ) 2023.06

性教育を身近な指導にパンフレットを用いるメリットと課題

米川葵, 渡辺絢子, 中島直美, 宮越郁子, 石郷友之, 加藤貴志, 片野唆敏, 小山雅之, 矢野俊之, 橋本暁佳, 古橋眞人

日本肺高血圧・肺循環学会学術集会抄録集 ( 日本肺高血圧・肺循環学会 ) 8回 153 - 153 2023.06

Three-Dimensional Spheroid Configurations and Cellular Metabolic Properties of Oral Squamous Carcinomas Are Possible Pharmacological and Pathological Indicators.

Sho Miyamoto, Nami Nishikiori, Tatsuya Sato, Megumi Watanabe, Araya Umetsu, Yuri Tsugeno, Fumihito Hikage, Takashi Sasaya, Hirotaka Kato, Kazuhiro Ogi, Masato Furuhashi, Hiroshi Ohguro, Akihiro Miyazaki

Cancers 15 ( 10 ) 2023.05 [International journal]

The objective of the current study was to elucidate the clinicopathological significance and appearance of in vitro three-dimension (3D) spheroid models of oral malignant tumors that were prepared from four pathologically different squamous cell carcinoma (OSCC; low-grade; SSYP and MO-1000, intermediate-grade; LEM2) and oral adenosquamous carcinoma (OASC; high-grade; Mesimo) obtained from patients with different malignant stages. To characterize the biological significance of these cell lines themselves, two-dimensional (2D) cultured cells were subjected to cellular metabolic analysis by a Seahorse bioanalyzer alongside the measurement of the cytotoxicity of cisplatin (CDDP). The appearance of their 3D spheroids was then observed by phase contrast microscopy, and both 2D and 3D cultured cells were subject to trypsin digestion and qPCR analysis of factors related to oncogenic signaling and other related analyses. ATP-linked respiration and proton leaking were significantly different among the four cell lines, and the malignant stages of these cultures were significantly associated with increased ATP-linked respiration and decreased proton leakage. Alternatively, the appearances of these 3D spheroids were also significantly diverse among them, and their differences increased in the order of LEM2, MO-1000, SSYP, and Mesimo. Interestingly, these orders were exactly the same in that the efficacies of CDDP-induced cytotoxicity increased in the same order. qPCR analysis indicated that the levels of expression of oncogenic signaling-related factors varied among these four cell lines, and the values for fibronectin and a key regulator of mitochondrial biogenesis, PGC-1α, were prominently elevated in cultures of the worst malignant Mesimo cells. In addition, although 0.25% trypsin-induced destruction was comparable among all four 2D cultured cells, the values for the 3D spheroids were also substantially varied among these cultures. The findings reported herein indicate that cellular metabolic functions and 3D spheroid architectures may be valuable and useful indicators for estimating the pathological and drug-sensitive aspects of OSCC and OASC malignancies.

IgA腎症での糸球体障害における脂肪酸結合蛋白4の関与についての検討

田中希尚, 茂庭仁人, 禾千絵子, 淺沼克彦, 鈴木祐介, 古橋眞人

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 65 ( 3 ) 271 - 271 2023.05

Chlamydia pneumoniae Lung Infection in Mice Induces Fatty Acid-Binding Protein 4-Dependent White Adipose Tissue Pathology.

Yusuke Kurihara, Nirwana Fitriani Walenna, Kazunari Ishii, Toshinori Soejima, Bin Chou, Michinobu Yoshimura, Ryo Ozuru, Akinori Shimizu, Ryota Itoh, Masato Furuhashi, Gökhan S Hotamisligil, Kenji Hiromatsu

Journal of immunology (Baltimore, Md. : 1950) 210 ( 8 ) 1086 - 1097 2023.04 [International journal]

Fatty acid-binding protein 4 (FABP4) is a critical immune-metabolic modulator, mainly expressed in adipocytes and macrophages, secreted from adipocytes in association with lipolysis, and plays essential pathogenic roles in cardiovascular and metabolic diseases. We previously reported Chlamydia pneumoniae infecting murine 3T3-L1 adipocytes and causing lipolysis and FABP4 secretion in vitro. However, it is still unknown whether C. pneumoniae intranasal lung infection targets white adipose tissues (WATs), induces lipolysis, and causes FABP4 secretion in vivo. In this study, we demonstrate that C. pneumoniae lung infection causes robust lipolysis in WAT. Infection-induced WAT lipolysis was diminished in FABP4-/- mice or FABP4 inhibitor-pretreated wild-type mice. Infection by C. pneumoniae in wild-type but not FABP4-/- mice induces the accumulation of TNF-α- and IL-6-producing M1-like adipose tissue macrophages in WAT. Infection-induced WAT pathology is augmented by endoplasmic reticulum (ER) stress/the unfolded protein response (UPR), which is abrogated by treatment with azoramide, a modulator of the UPR. C. pneumoniae lung infection is suggested to target WAT and induce lipolysis and FABP4 secretion in vivo via ER stress/UPR. FABP4 released from infected adipocytes may be taken up by other neighboring intact adipocytes or adipose tissue macrophages. This process can further induce ER stress activation and trigger lipolysis and inflammation, followed by FABP4 secretion, leading to WAT pathology. A better understanding of the role of FABP4 in C. pneumoniae infection-induced WAT pathology will provide the basis for rational intervention measures directed at C. pneumoniae infection and metabolic syndrome, such as atherosclerosis, for which robust epidemiologic evidence exists.

代謝異常関連脂肪肝疾患(MAFLD)は収縮期血圧の経時的な上昇と関連する線形混合効果モデルを用いた解析

遠藤圭佑, 森和真, 田中希尚, 佐藤達也, 大西浩文, 塙なぎさ, 古橋眞人

糖尿病 ( (一社)日本糖尿病学会 ) 66 ( Suppl.1 ) S - 264 2023.04

Downregulation of BCKDH Activity and Uncoupling from AMP Deaminase Underlie Substrate Inflexibility in Type 2 Diabetic Hearts(タイトル和訳中)

小川俊史, 神津英至, 長南新太, 清水将輝, 戸田悠貴, 大和田渉, 佐藤達也, 矢野俊之, 久野篤史, 丹野雅也, 三浦哲嗣, 古橋眞人

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 87回 OJ03 - 2 2023.03

心不全患者の骨粗鬆症を予測する新たなリスクスコアの開発と精度の検証

沼澤瞭, 片野唆敏, 長岡凌平, 藤澤佑輔, 山埜光太郎, 大堀克彦, 渡辺絢子, 石郷友之, 西川諒, 大和田渉, 永野伸卓, 藤戸健史, 小山雅之, 神津英至, 橋本暁佳, 矢野俊之, 古橋眞人

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 87回 CA1 - 2 2023.03

Impact of Iron Metabolism on Physical Function in Patients with Heart Failure(タイトル和訳中)

大堀克彦, 矢野俊之, 片野唆敏, 長岡凌平, 沼澤瞭, 山埜光太郎, 藤沢祐輔, 神津英至, 小山雅之, 永野伸卓, 藤戸健史, 西川諒, 大和田渉, 古橋眞人

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 87回 PJ038 - 3 2023.03

BCKDH活性のダウンレギュレーションとAMPデアミナーゼとのアンカップリングが2型糖尿病性心臓における基質柔軟性の欠如を引き起こす(Downregulation of BCKDH Activity and Uncoupling from AMP Deaminase Underlie Substrate Inflexibility in Type 2 Diabetic Hearts)

小川俊史, 神津英至, 長南新太, 清水将輝, 戸田悠貴, 大和田渉, 佐藤達也, 矢野俊之, 久野篤史, 丹野雅也, 三浦哲嗣, 古橋眞人

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 87回 OJ03 - 2 2023.03

3D Spheroid Configurations Are Possible Indictors for Evaluating the Pathophysiology of Melanoma Cell Lines.

Hiroshi Ohguro, Megumi Watanabe, Tatsuya Sato, Fumihito Hikage, Masato Furuhashi, Masae Okura, Tokimasa Hida, Hisashi Uhara

Cells 12 ( 5 ) 2023.02 [International journal]

To study the molecular mechanisms responsible for inducing the spatial proliferation of malignant melanomas (MM), three-dimension (3D) spheroids were produced from several MM cell lines including SK-mel-24, MM418, A375, WM266-4, and SM2-1, and their 3D architectures and cellular metabolisms were evaluated by phase-contrast microscopy and Seahorse bio-analyzer, respectively. Several transformed horizontal configurations were observed within most of these 3D spheroids, and the degree of their deformity was increased in the order: WM266-4, SM2-1, A375, MM418, and SK-mel-24. An increased maximal respiration and a decreased glycolytic capacity were observed within the lesser deformed two MM cell lines, WM266-4 and SM2-1, as compared with the most deformed ones. Among these MM cell lines, two distinct cell lines, WM266-4 and SK-mel-24, whose 3D appearances were the closest and farthest, respectively, from being horizontally circular-shaped, were subjected to RNA sequence analyses. Bioinformatic analyses of the differentially expressed genes (DEGs) identified KRAS and SOX2 as potential master regulatory genes for inducing these diverse 3D configurations between WM266-4 and SK-mel-24. The knockdown of both factors altered the morphological and functional characteristics of the SK-mel-24 cells, and in fact, their horizontal deformity was significantly reduced. A qPCR analysis indicated that the levels of several oncogenic signaling related factors, including KRAS and SOX2, PCG1α, extracellular matrixes (ECMs), and ZO1 had fluctuated among the five MM cell lines. In addition, and quite interestingly, the dabrafenib and trametinib resistant A375 (A375DT) cells formed globe shaped 3D spheroids and showed different profiles in cellular metabolism while the mRNA expression of these molecules that were tested as above were different compared with A375 cells. These current findings suggest that 3D spheroid configuration has the potential for serving as an indicator of the pathophysiological activities associated with MM.

TGF-β-3 Induces Different Effects from TGF-β-1 and -2 on Cellular Metabolism and the Spatial Properties of the Human Trabecular Meshwork Cells

Megumi Watanabe, Tatsuya Sato, Yuri Tsugeno, Megumi Higashide, Masato Furuhashi, Hiroshi Ohguro

International Journal of Molecular Sciences 2023.02 [Refereed]

Comparison of Phenotypes in Subcutaneous Fat and Perivascular Adipose Tissue Surrounding the Saphenous Vein in Coronary Artery Bypass Grafting

Takuma Mikami, Masato Furuhashi, Ryosuke Numaguchi, Itaru Hosaka, Akiko Sakai, Marenao Tanaka, Toshiro Ito, Toshiyuki Maeda, Taku Sakurada, Satoshi Muraki, Yousuke Yanase, Hiroshi Sato, Joji Fukada, Yukihiko Tamiya, Yutaka Iba, Nobuyoshi Kawaharada

Circulation Journal ( Japanese Circulation Society ) 2023.02

Thyroid-stimulating hormone高値は男性の新規CKD発症と関連する

遠藤圭佑, 田中希尚, 佐藤達也, 塙なぎさ, 大西浩文, 古橋眞人

日本内科学会雑誌 ( (一社)日本内科学会 ) 112 ( 臨増 ) 176 - 176 2023.02

Metabolic dysfunction-associated fatty liver disease is associated with an increase in systolic blood pressure over time: linear mixed-effects model analyses

Kazuma Mori, Marenao Tanaka, Itaru Hosaka, Takuma Mikami, Keisuke Endo, Nagisa Hanawa, Hirofumi Ohnishi, Masato Furuhashi

Hypertension Research ( Springer Science and Business Media LLC ) 2023.01

Glomerular expression and urinary excretion of fatty acid-binding protein 4 in IgA nephropathy

Marenao Tanaka, Norihito Moniwa, Chieko Nogi, Toshiki Kano, Megumi Matsumoto, Akiko Sakai, Takuto Maeda, Hideki Takizawa, Yayoi Ogawa, Katsuhiko Asanuma, Yusuke Suzuki, Masato Furuhashi

Journal of Nephrology ( Springer Science and Business Media LLC ) 2023.01

Unexpected Crosslinking Effects of a Human Thyroid Stimulating Monoclonal Autoantibody, M22, with IGF1 on Adipogenesis in 3T3L-1 Cells

Araya Umetsu, Tatsuya Sato, Megumi Watanabe, Yosuke Ida, Masato Furuhashi, Yuri Tsugeno, Hiroshi Ohguro

International Journal of Molecular Sciences 2023.01 [Refereed]

G-Protein-Coupled Receptors Mediate Modulations of Cell Viability and Drug Sensitivity by Aberrantly Expressed Recoverin 3 within A549 Cells

Hanae Ichioka, Yoshihiko Hirohashi, Tatsuya Sato, Masato Furuhashi, Megumi Watanabe, Yosuke Ida, Fumihito Hikage, Toshihiko Torigoe, Hiroshi Ohguro

International Journal of Molecular Sciences 24 ( 1 ) 2023.01 [Refereed] [International journal]

To elucidate the currently unknown molecular mechanisms responsible for the aberrant expression of recoverin (Rec) within cancerous cells, we examined two-dimensional (2D) and three-dimensional (3D) cultures of Rec-negative lung adenocarcinoma A549 cells which had been transfected with a plasmid containing human recoverin cDNA (A549 Rec) or an empty plasmid as a mock control (A549 MOCK). Using these cells, we measured cytotoxicity by several anti-tumor agents (2D), cellular metabolism including mitochondrial and glycolytic functions by a Seahorse bio-analyzer (2D), the physical properties, size and stiffness of the 3D spheroids, trypsin sensitivities (2D and 3D), and RNA sequencing analysis (2D). Compared with the A549 MOCK, the A549 Rec cells showed (1) more sensitivity toward anti-tumor agents (2D) and a 0.25% solution of trypsin (3D); (2) a metabolic shift from glycolysis to oxidative phosphorylation; and (3) the formation of larger and stiffer 3D spheroids. RNA sequencing analysis and bioinformatic analyses of the differentially expressed genes (DEGs) using Gene Ontology (GO) enrichment analysis suggested that aberrantly expressed Rec is most likely associated with several canonical pathways including G-protein-coupled receptor (GPCR)-mediated signaling and signaling by the cAMP response element binding protein (CREB). The findings reported here indicate that the aberrantly expressed Rec-induced modulation of the cell viability and drug sensitivity may be GPCR mediated.

Potential favorable action of sodium-glucose cotransporter-2 inhibitors on sudden cardiac death: a brief overview.

Tatsuya Sato, Hidemichi Kouzu, Toshiyuki Yano, Ichiro Sakuma, Masato Furuhashi, Noritsugu Tohse

Frontiers in cardiovascular medicine 10 1159953 - 1159953 2023 [International journal]

The primary pharmacological action of sodium-glucose co-transporter 2 (SGLT2) inhibitors is to inhibit the reabsorption of glucose and sodium ions from the proximal tubules of the kidney and to promote urinary glucose excretion. Notably, several clinical trials have recently demonstrated potent protective effects of SGLT2 inhibitors in patients with heart failure (HF) or chronic kidney disease (CKD), regardless of the presence or absence of diabetes. However, the impact of SGLT2 inhibitors on sudden cardiac death (SCD) or fatal ventricular arrhythmias (VAs), the pathophysiology of which is partly similar to that of HF and CKD, remains undetermined. The cardiorenal protective effects of SGLT2 inhibitors have been reported to include hemodynamic improvement, reverse remodeling of the failing heart, amelioration of sympathetic hyperactivity, correction of anemia and impaired iron metabolism, antioxidative effects, correction of serum electrolyte abnormalities, and antifibrotic effects, which may lead to prevent SCD and/or VAs. Recently, as possible direct cardiac effects of SGLT2 inhibitors, not only inhibition of Na+/H+ exchanger (NHE) activity, but also suppression of late Na+ current have been focused on. In addition to the indirect cardioprotective mechanisms of SGLT2 inhibitors, suppression of aberrantly increased late Na+ current may contribute to preventing SCD and/or VAs via restoration of the prolonged repolarization phase in the failing heart. This review summarizes the results of previous clinical trials of SGLT2 inhibitors for prevention of SCD, their impact on the indices of electrocardiogram, and the possible molecular mechanisms of their anti-arrhythmic effects.

Case report: Transient lactate elevation by intravenous insulin infusion therapy for diabetic ketoacidosis in a patient with mitochondrial DNA 3243 A > G mutation: A glycolysis rebooting syndrome?

Wataru Ohwada, Hidemichi Kouzu, Tatsuya Sato, Kahomi Sazawa, Azumi Matsui, Nobutaka Nagano, Masayuki Koyama, Noriko Ogasawara, Akifumi Takada, Toshiyuki Yano, Masato Furuhashi

Frontiers in cardiovascular medicine 10 1144925 - 1144925 2023 [International journal]

Mitochondrial disease, most cases of which are caused by mitochondrial DNA (mtDNA) mutation, is present with multiple phenotypes including diabetes mellitus, sensorineural hearing loss, cardiomyopathy, muscle weakness, renal dysfunction, and encephalopathy, depending on the degree of heteroplasmy. While mitochondria play an important role in intracellular glucose and lactate metabolism in insulin-sensitive tissues such as muscles, appropriate strategies for glycemic control have not yet been established in a patient with mitochondrial disease, which is often complicated by myopathy. Here, we describe the history of a 40-year-old man with mtDNA 3243A > G who had sensorineural hearing loss, cardiomyopathy, muscle wasting, and diabetes mellitus with stage 3 chronic kidney disease. He developed mild diabetic ketoacidosis (DKA) in the process of treatment for poor glycemic control with severe latent hypoglycemia. According to the standard therapy for DKA, he was treated with continuous intravenous insulin infusion therapy, which unexpectedly resulted in an abrupt and transient elevation in blood lactate levels without exacerbation of heart failure and kidney function. Since blood lactate levels are determined by the balance between lactate production and consumption, an abrupt and transient lactate elevation following intravenous insulin injection therapy may reflect not only enhanced glycolysis in insulin-sensitive tissues with mitochondrial dysfunction but also decreased lactate consumption in the sarcopenic skeletal muscle and failing heart. Intravenous insulin infusion therapy in patients with mitochondrial disease may unmask derangements of intracellular glucose metabolism in response to insulin signaling.

Elevated circulating level of β-aminoisobutyric acid (BAIBA) in heart failure patients with type 2 diabetes receiving sodium-glucose cotransporter 2 inhibitors

Satoshi Katano, Toshiyuki Yano, Hidemichi Kouzu, Ryohei Nagaoka, Ryo Numazawa, Kotaro Yamano, Yusuke Fujisawa, Katsuhiko Ohori, Nobutaka Nagano, Takefumi Fujito, Ryo Nishikawa, Wataru Ohwada, Masaki Katayose, Tatsuya Sato, Atsushi Kuno, Masato Furuhashi

Cardiovascular Diabetology ( Springer Science and Business Media LLC ) 21 ( 1 ) 2022.12

Abstract Aims The mechanism by which a sodium-glucose cotransporter inhibitor (SGLT2i) induces favorable effects on diabetes and cardiovascular diseases including heart failure (HF) remains poorly understood. Metabolomics including amino acid profiling enables detection of alterations in whole body metabolism. The aim of this study was to determine whether plasma amino acid profiles are modulated by SGLT2i use in HF patients with type 2 diabetes mellitus (T2DM). Methods We retrospectively examined 81 HF patients with T2DM (68 ± 11 years old; 78% male). Plasma amino acid concentrations in a fasting state after stabilization of HF were determined using ultraperformance liquid chromatography. To minimize potential selection bias in the retrospective analyses, the differences in baseline characteristics between patients receiving an SGLT2i and patients not receiving an SGLT2i were controlled by using an inverse probability of treatment weighting (IPTW)-adjusted analysis. Results Of amino acids measurable in the present assay, plasma β-aminoisobutyric acid (BAIBA), an exercise-induced myokine-like molecule also known as 3-aminoisobutyric acid or 3-amino-2-methyproponic acid, was detected in 77% of all patients and the proportion of patients in whom plasma BAIBA was detected was significantly higher in patients receiving an SGLT2i than in patients not receiving an SGLT2i (93% vs. 67%, p = 0.01). Analyses in patients in whom plasma BAIBA was detected showed that plasma BAIBA concentration was significantly higher in patients receiving an SGLT2i than in patients not receiving an SGLT2i (6.76 ± 4.72 vs. 4.56 ± 2.93 nmol/ml, p = 0.03). In multivariate logistic regression analyses that were adjusted for age and sex, SGLT2i use was independently associated with BAIBA detection. The independent association between BAIBA and SGLT2i use remained after inclusion of body mass index, HF with reduced ejection fraction, ischemic etiology, renal function, NT-proBNP, albumin, hemoglobin, and HbA1c into the Cox proportional hazards model. When the differences in baseline characteristics between patients receiving an SGLT2i and patients not receiving an SGLT2i were controlled by using an IPTW-adjusted analysis, least squares mean of plasma BAIBA concentration was significantly higher in patients receiving an SGLT2i than in patients not receiving an SGLT2i. Conclusion SGLT2i use is closely associated with increased circulating BAIBA concentration in HF patients with T2DM.

尿酸とCOVID-19の原因ウイルスSARS-CoV-2の宿主側受容体ACE2との関連についての検討

古橋眞人, 田中希尚, 東浦幸村, 森和真, 小山雅之, 大西浩文, 齋藤重幸, 島本和明

痛風と尿酸・核酸 ( (一社)日本痛風・尿酸核酸学会 ) 46 ( 2 ) 160 - 160 2022.12

血清尿酸値は女性において収縮期血圧の経時的な上昇と関連する線形混合効果モデルでの解析

森和真, 田中希尚, 東浦幸村, 小山雅之, 塙なぎさ, 大西浩文, 古橋眞人

痛風と尿酸・核酸 ( (一社)日本痛風・尿酸核酸学会 ) 46 ( 2 ) 166 - 166 2022.12

Addition of ROCK Inhibitors Alleviates Prostaglandin-Induced Inhibition of Adipogenesis in 3T3L-1 Spheroids.

Yosuke Ida, Tatsuya Sato, Araya Umetsu, Megumi Watanabe, Masato Furuhashi, Fumihito Hikage, Hiroshi Ohguro

Bioengineering (Basel, Switzerland) 9 ( 11 ) 2022.11 [International journal]

To elucidate the additive effects of the ROCK inhibitors (ROCK-i), ripasudil (Rip) and Y27632 on bimatoprost acid (BIM-A), a prostaglandin analog (PG), on adipose tissue, two- and three-dimensional (2D or 3D) cultures of 3T3-L1 cells, the most well characterized cells in the field of lipid research, were used. The cells were subjected to a variety of analyses including lipid staining, real-time cellular metabolic analysis, the mRNA expressions of genes related to adipogenesis and extracellular matrices (ECMs) as well as the sizes and physical properties of the 3D spheroids by a micro-squeezer. BIM-A induced strong inhibitory effects on most of the adipogenesis-related changes in the 2D and 3D cultured 3T3-L1 cells, including (1) the enlargement and softening of the 3D spheroids, (2) a dramatic enhancement in lipid staining and the expression of adipogenesis-related genes, and (3) a decrease in mitochondrial and glycolytic metabolic function. By adding ROCK-i to the BIM-A, most of these BIM-A-induced effects were cancelled. The collective findings reported herein suggest that ROCK-i eliminated the PG-induced suppression of adipogenesis in the 3T3-L1 cells, accompanied by the formation of enlarged 3D spheroids. Such effects of adding ROCK-i to a PG in preadipocytes on cellular properties appear to be associated with the suppression of PG-induced adverse effects, and provide additional insight into our understanding of lipid-related research.

The Selective α1 Antagonist Tamsulosin Alters ECM Distributions and Cellular Metabolic Functions of ARPE 19 Cells in a Concentration-Dependent Manner.

Yosuke Ida, Tatsuya Sato, Megumi Watanabe, Araya Umetsu, Yuri Tsugeno, Masato Furuhashi, Fumihito Hikage, Hiroshi Ohguro

Bioengineering (Basel, Switzerland) 9 ( 10 ) 2022.10 [International journal]

The purpose of the present study was to examine the effect of the selective α1 antagonist tamsulosin (TAM) on human retinal pigment epithelium cells, ARPE 19. Two-dimension (2D) and three-dimension (3D) cultured ARPE 19 cells were used in the following characterizations: (1) ultrastructure by scanning electron microscopy (SEM) (2D); (2) barrier functions by transepithelial electrical resistance (TEER) measurements, and FITC-dextran permeability (2D); (3) real time cellular metabolisms by Seahorse Bioanalyzer (2D); (4) physical properties, size and stiffness measurements (3D); and (5) expression of extracellular matrix (ECM) proteins, including collagen1 (COL1), COL4, COL6 and fibronectin (FN) by qPCR and immunohistochemistry (2D and 3D). TAM induced significant effects including: (1) alteration of the localization of the ECM deposits; (2) increase and decrease of the TEER values and FITC-dextran permeability, respectively; (3) energy shift from glycolysis into mitochondrial oxidative phosphorylation (OXPHOS); (4) large and stiffened 3D spheroids; and (5) down-regulations of the mRNA expressions and immune labeling of most ECM proteins in a concentration-dependent manner. However, in some ECM proteins, COL1 and COL6, their immunolabeling intensities were increased at the lowest concentration (1 μM) of TAM. Such a discrepancy between the gene expressions and immunolabeling of ECM proteins may support alterations of ECM localizations as observed by SEM. The findings reported herein indicate that the selective α1 antagonist, TAM, significantly influenced ECM production and distribution as well as cellular metabolism levels in a concentration-dependent manner.

Fatty liver index(FLI)の高値は虚血性心疾患の独立した予測因子である

森和真, 田中希尚, 東浦幸村, 塙なぎさ, 大西浩文, 古橋眞人

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 44回 60 - 60 2022.10

Fatty liver index(FLI)の高値は虚血性心疾患の独立した予測因子である

森和真, 田中希尚, 東浦幸村, 塙なぎさ, 大西浩文, 古橋眞人

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 44回 60 - 60 2022.10

FGF-2 enhances fibrogenetic changes in TGF-β2 treated human conjunctival fibroblasts.

Yuri Tsugeno, Masato Furuhashi, Tatsuya Sato, Megumi Watanabe, Araya Umetsu, Soma Suzuki, Yosuke Ida, Fumihito Hikage, Hiroshi Ohguro

Scientific reports 12 ( 1 ) 16006 - 16006 2022.09 [International journal]

The objective of the current study was to examine the effects of fibroblast growth factor-2 (FGF-2) on conjunctival fibrogenesis that was induced by the presence of transforming growth factor-β2 (TGF-β2). Two-dimension (2D) and three-dimension (3D) cultured human conjunctival fibroblasts (HconF) were used for this purpose. The 2D and 3D cultured HconF were characterized by transendothelial electrical resistance (TEER) and FITC dextran permeability measurements (2D), real-time metabolic analyses (2D), size and stiffness measurements (3D), and the mRNA expression of extracellular matrix molecules, their modulators, Tissue inhibitor of metalloproteinases and matrix metalloproteinases and ER-stress related genes (2D and 3D). FGF-2 significantly increased planar proliferation, as evidenced by TEER values and FITC dextran permeability, and shifted glucose metabolism to the energetic phenotype of 2D HconF cells, and the stiffness of the 3D spheroids, and these effects were further enhanced in the presence of TGF-β2. Analyses of the expression of possible candidate molecules involved in cell architecture and stress indicated that some additive effects caused by both factors were also recognized in some of these molecules. The findings reported herein indicate that the FGF-2, either along or additively with TGF- β2 increased the fibrogenetic changes on the plane as well as in the spatial space of HconF cells.

All Trans-Retinoic Acids Facilitate the Remodeling of 2D and 3D Cultured Human Conjunctival Fibroblasts.

Yuri Tsugeno, Tatsuya Sato, Megumi Watanabe, Megumi Higashide, Masato Furuhashi, Araya Umetsu, Soma Suzuki, Yosuke Ida, Fumihito Hikage, Hiroshi Ohguro

Bioengineering (Basel, Switzerland) 9 ( 9 ) 2022.09 [International journal]

Vitamin A derivative, all-trans-retinoic acid (ATRA), is known to be a potent regulator of the growth and differentiation of various types of cells. In the present study, the unidentified effects of ATRA on superficial and vertical spreading conjunctival scarring were examined. The study involved the use of two-dimensional (2D) and three-dimensional (3D) cultures of human conjunctival fibroblast (HconF) cells in the presence or absence of TGF-β2. The effects of ATRA (1 μM) on superficial or vertical spreading conjunctival scarring were evaluated by the barrier function by trans-endothelial electrical resistance (TEER) and FITC dextran permeability measurements and real-time metabolic analysis, as well as the physical properties, namely, the size and stiffness, of 3D spheroids, respectively. In addition, the expressions of several related molecules, including extracellular matrix (ECM) molecules, ECM modulators including a tissue inhibitor of metalloproteinases (TIMPs), matrix metalloproteinases (MMPs), and ER stress-related factors, were examined. ATRA significantly induced (1) an increase in TEER values and a decrease in FITC dextran permeability, respectively, in the 2D monolayers, and (2) relatively and substantially increased the size and stiffness, respectively, of the 3D spheroids. These ATRA-induced effects were further enhanced in the TGF-β2-treated cells, whereas the TGF-β2-induced enhancement in glycolytic capacity was canceled by the presence of ATRA. Consistent with these physical and morphological effects, the mRNA expressions of several molecules were significantly but differently induced between 2D and 3D cultures by ATRA, although the presence of TGF-β2 did not substantially affect these gene expression levels. The findings reported in this study indicate that ATRA may exacerbate both superficial and vertical conjunctival fibrosis spreading independently of TGF-β2-induced changes.

All-trans Retinoic Acids Synergistically and Beneficially Affect In Vitro Glaucomatous Trabecular Meshwork (TM) Models Using 2D and 3D Cell Cultures of Human TM Cells.

Megumi Watanabe, Tatsuya Sato, Yuri Tsugeno, Megumi Higashide, Masato Furuhashi, Araya Umetsu, Soma Suzuki, Yosuke Ida, Fumihito Hikage, Hiroshi Ohguro

International journal of molecular sciences 23 ( 17 ) 2022.08 [International journal]

We report herein on the effects of all-trans retinoic acid (ATRA) on two-dimensional (2D) and three-dimensional (3D) cultures of human trabecular meshwork (HTM) cells that were treated with transforming growth factor β2 (TGF-β2). In the presence of 5 ng/mL TGF-β2, the effects of ATRA on the following were observed: (1) the barrier function of the 2D HTM monolayers, as determined by trans-endothelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC) dextran permeability measurements; (2) a Seahorse cellular bio-metabolism analysis; (3) physical properties, including the size and stiffness, of 3D spheroids; (4) the gene expression of extracellular matrix (ECM) molecules, ECM modulators including tissue inhibitor of metalloproteinases (TIMPs), matrix metalloproteinases (MMPs), tight junction (TJ)-related molecules, and endoplasmic reticulum (ER)-stress-related factors. ATRA significantly inhibited the TGF-β2-induced increase in the TEER values and FITC dextran permeability of the 2D monolayers, while an ATRA monotreatment induced similar effects as TGF-β2. A real-time metabolic analysis revealed that ATRA significantly inhibited the TGF-β2-induced shift in metabolic reserve from mitochondrial oxidative phosphorylation to glycolysis in 2D HTM cells, whereas ATRA alone did not induce significant metabolic changes. In contrast, ATRA induced the formation of substantially downsized and softer 3D spheroids in the absence and presence of TGF-β2. The different effects induced by ATRA toward 2D and 3D HTM cells were also supported by the qPCR analysis of several proteins as above. The findings reported here indicate that ATRA may induce synergistic and beneficial effects on TGF-β2-treated 2D- and 3D-cultured HTM cells; those effects varied significantly between the 2D and 3D cultures.

Brimonidine Modulates the ROCK1 Signaling Effects on Adipogenic Differentiation in 2D and 3D 3T3-L1 Cells

Araya Umetsu, Yosuke Ida, Tatsuya Sato, Megumi Watanabe, Yuri Tsugeno, Masato Furuhashi, Fumihito Hikage, Hiroshi Ohguro

Bioengineering 2022.07 [Refereed]

An α2-Adrenergic Agonist, Brimonidine, Beneficially Affects the TGF-β2-Treated Cellular Properties in an In Vitro Culture Model

Megumi Watanabe, Tatsuya Sato, Yuri Tsugeno, Megumi Higashide, Masato Furuhashi, Araya Umetsu, Soma Suzuki, Yosuke Ida, Fumihito Hikage, Hiroshi Ohguro

Bioengineering ( {MDPI} {AG} ) 9 ( 7 ) 310 - 310 2022.07 [Refereed]

Human Trabecular Meshwork (HTM) Cells Treated with TGF-β2 or Dexamethasone Respond to Compression Stress in Different Manners.

Megumi Watanabe, Tatsuya Sato, Yuri Tsugeno, Araya Umetsu, Soma Suzuki, Masato Furuhashi, Yosuke Ida, Fumihito Hikage, Hiroshi Ohguro

Biomedicines 10 ( 6 ) 2022.06 [International journal]

To characterize our recently established in vitro glaucomatous human trabecular meshwork (HTM) models using dexamethasone (DEX)- or TGF-β2-treated HTM cells, (1) two-dimensional (2D) cultured HTM cells were characterized by means of the real-time cellular metabolism analysis using a Seahorse analyzer, and (2) the effects of mechanical compression stresses toward the three-dimensional (3D) HTM spheroids were evaluated by analyzing the gene expression of several ECM proteins, inflammatory cytokines, and ER stress-related factors of those 3D HTM spheroid models. The results indicated that (1) the real-time cellular metabolism analysis indicated that TGF-β2 significantly induced an energy shift from mitochondrial oxidative phosphorylation (OXPHOS) into glycolysis, and DEX induced similar but lesser effects. In contrast, ROCK2 inhibition by KD025 caused a substantial reverse energy shift from glycolysis into OXPHOS. (2) Upon direct compression stresses toward the untreated control 3D HTM spheroids, a bimodal fluctuation of the mRNA expressions of ECM proteins was observed for 60 min, that is, initial significant upregulation (0-10 min) and subsequent downregulation (10-30 min) followed by another upregulation (30-60 min); those of inflammatory cytokines and ER stress-related factors were also bimodally changed. However, such compression stresses for 30 min toward TGF-β2- or DEX-treated 3D HTM spheroids induced downregulation of most of those of inflammatory cytokines and ER stress-related factors in addition to upregulation of COL1 and downregulation of FN. The findings presented herein indicate that (1) OXPHOS of the HTM cells was decreased or increased by TGF-β2 or DEX stimulation or ROCK2 inhibition, and (2) mechanical compression stresses toward 3D HTM spheroids may replicate acute, subacute, and chronic HTM models affected by elevated intraocular pressures.

Metabolic dysfunction-associated fatty liver disease predicts new onset of chronic kidney disease better than does fatty liver or nonalcoholic fatty liver disease.

Marenao Tanaka, Kazuma Mori, Satoko Takahashi, Yukimura Higashiura, Hirofumi Ohnishi, Nagisa Hanawa, Masato Furuhashi

Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association 2022.05 [International journal]

BACKGROUND: Possible associations of chronic kidney disease (CKD) with fatty liver (FL) and nonalcoholic fatty liver disease (NAFLD) have recently been focused on. Metabolic dysfunction-associated fatty liver disease (MAFLD), defined as FL with overweight/obesity, type 2 diabetes mellitus or metabolic abnormalities, has been proposed as a new feature of chronic liver disease. However, the relationship between MAFLD and new onset of CKD has not been fully addressed. METHODS: We investigated the associations of FL, NAFLD and MAFLD with the development of CKD, defined as estimated glomerular filtration rate (eGFR) < 60 mL/min/1.73m2 or positive for urinary protein, over a 10-year period in 28 890 Japanese subjects who received annual health examinations. After exclusion of subjects with no data for abdominal ultrasonography and subjects with CKD at baseline, a total of 13 159 subjects (men/women: 8581/4578, mean age: 48 years) were recruited. RESULTS: The prevalences of FL, NAFLD and MAFLD were 34.6% (men/women: 45.1%/15.1%), 32.8% (men/women: 42.7%/14.5%) and 32.3% (men/women: 42.4%/13.4%), respectively. During the 10-year follow-up period, 2163 subjects (men/women: 1475/688) had new onset of CKD. Multivariable Cox proportional hazard model analyses showed that MAFLD (hazard ratio [95% confidence interval]: 1.12 [1.02-1.26], p = 0.027), but not FL or NAFLD, was an independent risk factor for new onset of CKD after adjustment of age, sex, eGFR, current smoking habit, ischemic heart disease, diabetes mellitus, overweight/obesity, hypertension and dyslipidemia. The addition of MAFLD (continuous net reclassification improvement [NRI]: 0.154, integrated discrimination improvement [IDI]: 0.0024) to traditional risk factors without metabolic abnormalities significantly improved the discriminatory capacity better than did the addition of FL (NRI: 0.138, IDI: 0.0018) or NAFLD (NRI: 0.132, IDI: 0.0017). CONCLUSIONS: MAFLD is modestly and independently associated with new onset of CKD and predicts the risk for development of CKD better than does FL or NAFLD.

High fatty liver index is an independent predictor of ischemic heart disease during a 10-year period in a Japanese population.

Kazuma Mori, Marenao Tanaka, Yukimura Higashiura, Nagisa Hanawa, Hirofumi Ohnishi, Masato Furuhashi

Hepatology research : the official journal of the Japan Society of Hepatology 52 ( 8 ) 687 - 698 2022.05 [International journal]

AIM: Fatty liver index (FLI), which is calculated by using body mass index, waist circumference and levels of γ-glutamyl transferase and triglycerides, is a validated surrogate marker of nonalcoholic fatty liver disease. We retrospectively investigated the relationship between FLI and the development of ischemic heart disease (IHD) during a 10-year period. METHODS: Among subjects who received annual health checkups (n = 28 990), a total of 18 851 subjects (men/women: 11 659/7192) were enrolled after exclusion of subjects with missing data and those with IHD at baseline. RESULTS: FLI at baseline was significantly higher in men than in women. During the 10-year period, 450 men (3.9%) and 123 women (1.7%) had new onset of IHD determined by a self-reported questionnaire survey. Multivariable Cox proportional hazard models with a restricted cubic spline showed that hazard risk (HR) for the development of IHD increased with a higher FLI at baseline after adjustment of age, sex, current smoking habit, family history of IHD and diagnosis of diabetes mellitus, hypertension, dyslipidemia and chronic kidney disease at baseline. There was no significant interaction between FLI and sex for the adjusted HR. When divided by tertiles of FLI at baseline (T1∼T3), the adjusted risk for development of IHD in the T3 group (HR [95% confidence interval]: 1.34 [1.05-1.71]) was significantly higher than that in the T1 group as the reference. The addition of FLI into traditional risk factors for IHD significantly improved the discriminatory capability. CONCLUSIONS: A high level of FLI is an independent predictor of new onset of IHD during a 10-year period.

Hypoxia Differently Affects TGF-β2-Induced Epithelial Mesenchymal Transitions in the 2D and 3D Culture of the Human Retinal Pigment Epithelium Cells.

Soma Suzuki, Tatsuya Sato, Megumi Watanabe, Megumi Higashide, Yuri Tsugeno, Araya Umetsu, Masato Furuhashi, Yosuke Ida, Fumihito Hikage, Hiroshi Ohguro

International journal of molecular sciences 23 ( 10 ) 2022.05 [International journal]

The hypoxia associated with the transforming growth factor-β2 (TGF-β2)-induced epithelial mesenchymal transition (EMT) of human retinal pigment epithelium (HRPE) cells is well recognized as the essential underlying mechanism responsible for the development of proliferative retinal diseases. In vitro, three-dimensional (3D) models associated with spontaneous O2 gradients can be used to recapitulate the pathological levels of hypoxia to study the effect of hypoxia on the TGF-β2-induced EMT of HRPE cells in detail, we used two-dimensional-(2D) and 3D-cultured HRPE cells. TGF-β2 and hypoxia significantly and synergistically increased the barrier function of the 2D HRPE monolayers, as evidenced by TEER measurements, the downsizing and stiffening of the 3D HRPE spheroids and the mRNA expression of most of the ECM proteins. A real-time metabolic analysis indicated that TGF-β2 caused a decrease in the maximal capacity of mitochondrial oxidative phosphorylation in the 2D HRPE cells, whereas, in the case of 3D HRPE spheroids, TGF-β2 increased proton leakage. The findings reported herein indicate that the TGF-β2-induced EMT of both the 2D and 3D cultured HRPE cells were greatly modified by hypoxia, but during these EMT processes, the metabolic plasticity was different between 2D and 3D HRPE cells, suggesting that the mechanisms responsible for the EMT of the HRPE cells may be variable during their spatial spreading.

Reply to the comments of Naharci <i>et al</i> . on “Circulating level of fatty acid‐binding protein 4 is an independent predictor of metabolic dysfunction‐associated fatty liver disease in middle‐aged and elderly individuals”

Marenao Tanaka, Masato Furuhashi

Journal of Diabetes Investigation ( Wiley ) 13 ( 5 ) 928 - 929 2022.05

代謝異常関連脂肪肝疾患(MAFLD)とCKD発症リスクの関連についての検討

田中希尚, 高橋聖子, 森和真, 東浦幸村, 大西浩文, 塙なぎさ, 古橋眞人

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 64 ( 3 ) 223 - 223 2022.05

代謝異常関連脂肪肝疾患(MAFLD)とCKD発症リスクの関連についての検討

田中希尚, 高橋聖子, 森和真, 東浦幸村, 大西浩文, 塙なぎさ, 古橋眞人

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 64 ( 3 ) 223 - 223 2022.05

Increased LDL cholesterol level is associated with deterioration of renal function in males

Arata Osanami, Marenao Tanaka, Masato Furuhashi, Hirofumi Ohnishi, Nagisa Hanawa, Tomohisa Yamashita, Norihito Moniwa, Tetsuji Miura

Clinical Kidney Journal ( Oxford University Press (OUP) ) 2022.04

Abstract Background Relationships between levels of serum lipid fractions and the time course of renal function are discrepant in the literature. Here we examined this issue by analyses of healthy subjects in a cohort. Methods Of all subjects who received health examinations at Keijinkai Maruyama Clinic, Sapporo in 2006, subjects with hypertension, diabetes mellitus or chronic kidney disease (CKD) and those taking medication for dyslipidemia were excluded, and a total of 5,586 subjects (male/female: 3,563/2,023, mean age: 43 ± 8 years) were followed for 10 years. Results Linear mixed effect models showed that baseline low-density lipoprotein (LDL)-cholesterol level was negatively associated with estimated glomerular filtration rate (eGFR) during the 10-year follow-up period after adjustment of confounders. Interactions between the follow-up year and baseline level of LDL-cholesterol or high-density lipoprotein (HDL)-cholesterol for eGFR values during the follow-up period were significant in males but not in females. There were no significant interactions for eGFR between the follow-up year and baseline levels of total cholesterol, triglycerides, or HDL-cholesterol/triglycerides ratio. During the follow-up period, 346 males and 223 females developed CKD. When male subjects were divided into subgroups according to tertiles of baseline levels of LDL-cholesterol, the adjusted risk for CKD in the third tertial group was significantly higher than that in the first tertile group as a reference (hazard ratio [95% confidence interval]: 1.39 [1.02-1.90], p = 0.035). Such a difference was not observed for LDL-cholesterol tertiles in females or HDL-cholesterol tertiles in both sexes. Conclusions A high LDL-cholesterol level may be a risk factor of new-onset CKD in apparently healthy males.

High fibrosis-4 (FIB-4) index predicts new onset of ischemic heart disease during a 10-year period in a general population

Yukimura Higashiura, Marenao Tanaka, Kazuma Mori, Takuma Mikami, Itaru Hosaka, Hirofumi Ohnishi, Nagisa Hanawa, Masato Furuhashi

European Heart Journal Open ( Oxford University Press (OUP) ) 2022.04

Abstract Aims The fibrosis-4 (FIB-4) index, calculated by using age, platelet count and levels of aspartate aminotransferase and alanine aminotransferase, is a noninvasive indicator for detection of liver fibrosis. Advanced hepatic fibrosis is associated with morbidity and mortality in patients with nonalcoholic fatty liver disease. However, the relationship between liver fibrosis and the development of ischemic heart disease (IHD) has not fully been addressed. Methods and Results We investigated the association between FIB-4 index and new onset of IHD during a 10-year period in a general population of subjects who received annual health examinations (n = 28,990). After exclusion of subjects with missing data and those with a history of IHD at baseline, a total of 13,448 subjects (men/women: 8,774/4,674, mean age: 48 years) were included. During the 10-year period, 378 men (4.3%) and 77 women (1.6%) had new onset of IHD. Multivariable Cox proportional hazard models with a restricted cubic spline showed that hazard risk for the development of IHD increased with a higher FIB-4 index at baseline after adjustment of age, sex, fatty liver (FL) determined by ultrasonography, estimated glomerular filtration rate, habits of current smoking and alcohol drinking, family history of IHD, and diagnosis of hypertension, diabetes mellitus and dyslipidemia. When divided by FL, FIB-4 index was an independent predictor for the development of IHD in subjects with FL but not in those without FL. The addition of FIB-4 index to traditional risk factors for IHD significantly improved the discriminatory capability. Conclusion A high level of FIB-4 index predicts new onset of IHD during a 10-year period.

Fibrosis-4(FIB-4)Indexは新規の虚血性心疾患発症の独立した予測因子である

東浦幸村, 古橋眞人, 田中希尚, 森和真, 高橋聖子, 大西浩文, 塙なぎさ

糖尿病 ( (一社)日本糖尿病学会 ) 65 ( Suppl.1 ) S - 187 2022.04

Plasma Tsukushi Concentration Is Associated with High Levels of Insulin and FGF21 and Low Level of Total Cholesterol in a General Population without Medication.

Masato Furuhashi, Yukimura Higashiura, Akiko Sakai, Masayuki Koyama, Marenao Tanaka, Shigeyuki Saitoh, Kazuaki Shimamoto, Hirofumi Ohnishi

Metabolites 12 ( 3 ) 2022.03 [International journal]

Tsukushi (TSK) is a member of the small leucine-rich proteoglycan family that controls developmental processes and organogenesis. TSK was also identified as a new hepatokine, which is mainly expressed in the liver, and is secreted by hepatocytes, to regulate energy and glycolipid metabolism in response to nonalcoholic fatty liver disease. However, the role of plasma TSK, especially its role in the general population, has not been fully addressed. We investigated the associations between plasma TSK concentration and several metabolic markers, including fibroblast growth factor 21 (FGF21), a hepatokine, and adiponectin, an adipokine, in 253 subjects (men/women: 114/139) with no medication in the Tanno-Sobetsu Study, which employed a population-based cohort. There was no significant sex difference in plasma TSK concentration, and the level was positively correlated with the fatty liver index (FLI) (r = 0.131, p = 0.038), levels of insulin (r = 0.295, p < 0.001) and levels of FGF21 (r = 0.290, p < 0.001), and was negatively correlated with the total cholesterol level (r = -0.124, p = 0.049). There was no significant correlation between the TSK level and body mass index, waist circumference, adiponectin, high-density lipoprotein cholesterol or total bile acids. The multivariable regression analysis showed that high levels of insulin and FGF21 and a low level of total cholesterol were independent determinants of plasma TSK concentration, after adjustment for age, sex and FLI. In conclusion, plasma TSK concentration is independently associated with high levels of insulin and FGF21, a hepatokine, and a low level of total cholesterol, but not with adiposity and adiponectin, in a general population of subjects who have not taken any medications.

日本人におけるFLIによるNAFLDの予測

高橋聖子, 田中希尚, 東浦幸村, 塙なぎさ, 大西浩文, 古橋眞人

日本内科学会雑誌 ( (一社)日本内科学会 ) 111 ( Suppl. ) 198 - 198 2022.02

日本人におけるFLIによるNAFLDの予測

高橋聖子, 田中希尚, 東浦幸村, 塙なぎさ, 大西浩文, 古橋眞人

日本内科学会雑誌 ( (一社)日本内科学会 ) 111 ( Suppl. ) 198 - 198 2022.02

A case of membranous nephropathy secondary to asymptomatic Graves’ disease

Norihito Moniwa, Yu Shioya, Yufu Gocho, Satoko Takahashi, Marenao Tanaka, Masato Furuhashi, Setsuko Kuroda, Taketsugu Hama, Yuko Shima, Yayoi Ogawa, Tetsuji Miura

CEN Case Reports ( Springer Science and Business Media LLC ) 2022.01

Differential Effects of DPP-4 Inhibitors, Anagliptin and Sitagliptin, on PCSK9 Levels in Patients with Type 2 Diabetes Mellitus who are Receiving Statin Therapy.

Masato Furuhashi, Ichiro Sakuma, Takeshi Morimoto, Yukimura Higashiura, Akiko Sakai, Megumi Matsumoto, Mio Sakuma, Michio Shimabukuro, Takashi Nomiyama, Osamu Arasaki, Koichi Node, Shinichiro Ueda

Journal of atherosclerosis and thrombosis 29 ( 1 ) 24 - 37 2022.01 [Domestic journal]

AIM: Proprotein convertase subtilisin/kexin type 9 (PCSK9) degrades the low-density lipoprotein (LDL) receptor, leading to hypercholesterolemia and cardiovascular risk. Treatment with a statin leads to a compensatory increase in circulating PCSK9 level. Anagliptin, a dipeptidyl peptidase-4 (DPP-4) inhibitor, was shown to decrease LDL cholesterol (LDL-C) levels to a greater extent than that by sitagliptin, another DPP-4 inhibitor, in the Randomized Evaluation of Anagliptin versus Sitagliptin On low-density lipoproteiN cholesterol in diabetes (REASON) trial. We investigated PCSK9 concentration in type 2 diabetes mellitus (T2DM) and the impact of treatment with anagliptin or sitagliptin on PCSK9 level as a sub-analysis of the REASON trial. METHODS: PCSK9 concentration was measured at baseline and after 52 weeks of treatment with anagliptin (n=122) or sitagliptin (n=128) in patients with T2DM who were receiving statin therapy. All of the included patients had been treated with a DPP-4 inhibitor prior to randomization. RESULTS: Baseline PCSK9 level was positively, but not significantly, correlated with LDL-C and was independently associated with platelet count and level of triglycerides. Concomitant with reduction of LDL-C, but not hemoglobin A1c (HbA1c), by anagliptin, PCSK9 level was significantly increased by treatment with sitagliptin (218±98 vs. 242±115 ng/mL, P=0.01), but not anagliptin (233±97 vs. 250±106 ng/mL, P=0.07). CONCLUSIONS: PCSK9 level is independently associated with platelet count and level of triglycerides, but not LDL-C, in patients with T2DM. Anagliptin reduces LDL-C level independent of HbA1c control in patients with T2DM who are on statin therapy possibly by suppressing excess statin-mediated PCSK9 induction and subsequent degradation of the LDL receptor.

血清尿酸値は女性において収縮期血圧の経時的な上昇と関連する線形混合効果モデルでの解析

森和真, 田中希尚, 東浦幸村, 小山雅之, 塙なぎさ, 大西浩文, 古橋眞人

日本痛風・尿酸核酸学会総会プログラム・抄録集 ( (一社)日本痛風・尿酸核酸学会 ) 55回 86 - 86 2022.01

尿酸とCOVID-19の原因ウイルスSARS-CoV-2の宿主側受容体ACE2との関連についての検討

古橋眞人, 田中希尚, 東浦幸村, 森和真, 小山雅之, 大西浩文, 齋藤重幸, 島本和明

日本痛風・尿酸核酸学会総会プログラム・抄録集 ( (一社)日本痛風・尿酸核酸学会 ) 55回 80 - 80 2022.01

血清尿酸値は女性において収縮期血圧の経時的な上昇と関連する線形混合効果モデルでの解析

森和真, 田中希尚, 東浦幸村, 小山雅之, 塙なぎさ, 大西浩文, 古橋眞人

日本痛風・尿酸核酸学会総会プログラム・抄録集 ( (一社)日本痛風・尿酸核酸学会 ) 55回 86 - 86 2022.01

尿酸とCOVID-19の原因ウイルスSARS-CoV-2の宿主側受容体ACE2との関連についての検討

古橋眞人, 田中希尚, 東浦幸村, 森和真, 小山雅之, 大西浩文, 齋藤重幸, 島本和明

日本痛風・尿酸核酸学会総会プログラム・抄録集 ( (一社)日本痛風・尿酸核酸学会 ) 55回 80 - 80 2022.01

Circulating level of FABP4 is an independent predictor of metabolic dysfunction‐associated fatty liver disease in middle‐aged and elderly individuals

Marenao Tanaka, Satoko Takahashi, Yukimura Higashiura, Akiko Sakai, Masayuki Koyama, Shigeyuki Saitoh, Kazuaki Shimamoto, Hirofumi Ohnishi, Masato Furuhashi

Journal of Diabetes Investigation ( Wiley ) 2021.12

Fatty liver index is independently associated with deterioration of renal function during a 10-year period in healthy subjects

Satoko Takahashi, Marenao Tanaka, Masato Furuhashi, Norihito Moniwa, Masayuki Koyama, Yukimura Higashiura, Arata Osanami, Yufu Gocho, Hirofumi Ohnishi, Keita Numata, Takashi Hisasue, Nagisa Hanawa, Tetsuji Miura

Scientific Reports ( Springer Science and Business Media {LLC} ) 11 ( 1 ) 2021.12

AbstractA potential link between chronic kidney disease (CKD) and non-alcoholic fatty liver disease (NAFLD) has been suggested. We investigated the relationship between fatty liver index (FLI), a noninvasive and simple predictor of NAFLD, and the development of CKD defined as estimated glomerular filtration rate < 60 mL/min/1.73 m2 or positive for urinary protein during a 10-year follow-up period in subjects who received annual health examinations (n = 28,890). After exclusion of CKD at baseline, a total of 14,163 subjects (male/female: 9077/5086) were recruited. During the 10-year period, 1458 males (16.1%) and 737 females (14.5%) had new onset of CKD. Multivariable Cox proportional hazard models with a restricted cubic spline showed that hazard ratios (HRs) of CKD development increased with a higher FLI at baseline in both males and females after adjustment of confounders. When divided by tertiles of FLI level at baseline (T1 ~ T3), the adjusted risk of CKD development in the T3 group (HR [95% confidence interval], male/female: 1.33 [1.16–1.54]/1.33 [1.08–1.63]) was significantly higher than that in both sexes in the T1 group as the reference. The addition of FLI into traditional risk factors significantly improved the discriminatory capability for predicting CKD. In conclusion, a high level of FLI predicts the development of CKD in both sexes in a general population.

High level of fatty liver index predicts new onset of diabetes mellitus during a 10-year period in healthy subjects

Yukimura Higashiura, Masato Furuhashi, Marenao Tanaka, Satoko Takahashi, Masayuki Koyama, Hirofumi Ohnishi, Keita Numata, Takashi Hisasue, Nagisa Hanawa, Norihito Moniwa, Kazufumi Tsuchihashi, Tetsuji Miura

Scientific Reports ( Springer Science and Business Media {LLC} ) 11 ( 1 ) 2021.12

AbstractFatty liver index (FLI), a predictor of nonalcoholic fatty liver disease, has been reported to be associated with several metabolic disorders. This study aimed to evaluate the relationship between FLI and new onset of diabetes mellitus (DM). We investigated the association of FLI with new onset of DM during a 10-year period in subjects who received annual health examinations (n = 28,990). After exclusion of subjects with DM at baseline and those with missing data, a total of 12,290 subjects (male/female: 7925/4365) who received health examinations were recruited. FLI was significantly higher in males than in females. During the 10-year period, DM was developed in 533 males (6.7%) and 128 females (2.9%). Multivariable Cox proportional hazard models with a restricted cubic spline showed that the risk of new onset of DM increased with a higher FLI at baseline in both sexes after adjustment of age, fasting plasma glucose, habits of alcohol drinking and current smoking, family history of DM and diagnosis of hypertension and dyslipidemia at baseline. When the subjects were divided into subgroups according to tertiles of FLI level at baseline (T1–T3) in the absence and presence of impaired fasting glucose (IFG), hazard ratios after adjustment of the confounders gradually increased from T1 to T3 and from the absence to presence of IFG in both male and female subjects. In conclusion, a high level of FLI predicts new onset of DM in a general population of both male and female individuals.

Detection of significantly high vitreous concentrations of fatty acid-binding protein 4 in patients with proliferative diabetic retinopathy

Kaku Itoh, Masato Furuhashi, Yosuke Ida, Hiroshi Ohguro, Megumi Watanabe, Soma Suzuki, Fumihito Hikage

Scientific Reports ( Springer Science and Business Media {LLC} ) 11 ( 1 ) 2021.12

AbstractThe fatty acid-binding protein4 (FABP4) and vascular endothelial growth factor A (VEGFA) play key roles in the metabolic and cardiovascular diseases, and proliferative diabetic retinopathy (PDR), respectively. To identify FABP4 in vitreous fluid in PDR, vitreous concentrations of FABP4 (V-FABP4) and VEGFA (V-VEGFA) from PDR (n = 20) and non-PDR (n = 20) patients were determined by Enzyme-Linked ImmunoSorbent Assays. The data, which included height and weight, systemic blood pressures, several blood biochemical parameters and blood flow at the optic nerve head (ONH) by laser speckle flowgraphy (LSFG) were collected. The levels of V-FABP4 and V-VEGFA were significantly higher in PDR patients than in non-PDR patients (P < 0.001) with a high positive correlation (r = 0.72, P < 0.001) between them. The findings were not affected by body mass index values and the presence of vitreous hemorrhaging. Among the clinical parameters, V-FABP4 correlated positively with creatinine and negatively with age and aspartate transaminase (AST) levels, while V-VEGFA correlated positively with fasting plasma glucose and hemoglobin A1c (HbA1c) levels but negatively with AST. Multiple regression analyses indicated that V-VEGFA, or V-FABP4, AST and HbA1c were independent predictors of V-FABP4 or V-VEGFA, respectively. Both were negatively correlated, but more evident in V-FABP4, with the ONH ocular blood flow.

Elevated circulating FABP4 concentration predicts cardiovascular death in a general population: a 12-year prospective study

Norie Saito, Masato Furuhashi, Masayuki Koyama, Yukimura Higashiura, Hiroshi Akasaka, Marenao Tanaka, Norihito Moniwa, Hirofumi Ohnishi, Shigeyuki Saitoh, Nobuyuki Ura, Kazuaki Shimamoto, Tetsuji Miura

Scientific Reports ( Springer Science and Business Media {LLC} ) 11 ( 1 ) OJ16 - 4 2021.12

AbstractFatty acid-binding protein 4 (FABP4) is secreted from adipose tissue and acts as an adipokine, and an elevated circulating FABP4 level is associated with metabolic disorders and atherosclerosis. However, little is known about the causal link between circulating FABP4 level and mortality in a general population. We investigated the relationship between FABP4 concentration and mortality including cardiovascular death during a 12-year period in subjects of the Tanno-Sobetsu Study, a population-based cohort (n = 721, male/female: 302/419). FABP4 concentration at baseline was significantly higher in female subjects than in male subjects. All-cause death occurred in 123 (male/female: 74/49) subjects, and 34 (male/female: 20/14) and 42 (male/female: 26/16) subjects died of cardiovascular events and cancer, respectively. When divided into 3 groups according to tertiles of FABP4 level at baseline by sex (T1–T3), Kaplan–Meier survival curves showed that there were significant differences in rates of all-cause death and cardiovascular death, but not cancer death, among the groups. Multivariable Cox proportional hazard model analysis with a restricted cubic spline showed that hazard ratio (HR) for cardiovascular death, but not that for all-cause death, significantly increased with a higher FABP4 level at baseline after adjustment of age and sex. The risk of cardiovascular death after adjustment of age, sex, body mass index and levels of brain natriuretic peptide and high-sensitivity C-reactive protein in the 3rd tertile (T3) group (HR: 4.96, 95% confidence interval: 1.20–22.3) was significantly higher than that in the 1st tertile (T1) group as the reference. In conclusion, elevated circulating FABP4 concentration predicts cardiovascular death in a general population.

Serum uric acid level is associated with an increase in systolic blood pressure over time in female subjects: Linear mixed-effects model analyses

Kazuma Mori, Masato Furuhashi, Marenao Tanaka, Yukimura Higashiura, Masayuki Koyama, Nagisa Hanawa, Hirofumi Ohnishi

Hypertension Research ( Springer Science and Business Media LLC ) 2021.11

Pan-rock and rock2 inhibitors affect dexamethasone-treated 2d-and 3d-cultured human trabecular meshwork (Htm) cells in opposite manners

Megumi Watanabe, Yosuke Ida, Masato Furuhashi, Yuri Tsugeno, Fumihito Hikage, Hiroshi Ohguro

Molecules ( MDPI ) 26 ( 21 ) 2021.11

Effects of a pan-ROCK-inhibitor, ripasudil (Rip), and a ROCK2 inhibitor, KD025 on dex-amethasone (DEX)-treated human trabecular meshwork (HTM) cells as a model of steroid-induced glaucoma were investigated. In the presence of Rip or KD025, DEX-treated HTM cells were subjected to permeability analysis of 2D monolayer by transendothelial electrical resistance (TEER) and FITC– dextran permeability, physical properties, size and stiffness analysis (3D), and qPCR of extracellular matrix (ECM), and their modulators. DEX resulted in a significant increase in the permeability, as well as a large and stiff 3D spheroid, and those effects were inhibited by Rip. In contrast, KD025 exerted opposite effects on the physical properties (down-sizing and softening). Furthermore, DEX induced several changes of gene expressions of ECM and their modulators were also modulated differently by Rip and KD025. The present findings indicate that Rip and KD025 induced opposite effects toward 2D and 3D cell cultures of DEX-treated HTM cells.

Autotaxin may have lysophosphatidic acid-unrelated effects on three-dimension (3d) cultured human trabecular meshwork (htm) cells

Megumi Watanabe, Masato Furuhashi, Yuri Tsugeno, Yosuke Ida, Fumihito Hikage, Hiroshi Ohguro

International Journal of Molecular Sciences ( MDPI ) 22 ( 21 ) 2021.11

Purpose: The objective of the current study was to evaluate the effects of the autotaxin (ATX)–lysophosphatidic acid (LPA) signaling axis on the human trabecular meshwork (HTM) in two-dimensional (2D) and three-dimensional (3D) cultures of HTM cells. Methods: The effects were characterized by transendothelial electrical resistance (TEER) and FITC-dextran permeability (2D), measurements of size and stiffness (3D), and the expression of several genes, including extracellular matrix (ECM) molecules, their modulators, and endoplasmic reticulum (ER) stress-related factors. Results: A one-day exposure to 200 nM LPA induced significant down-sizing effects of the 3D HTM spheroids, and these effects were enhanced slightly on longer exposure. The TEER and FITC-dextran permeability data indicate that LPA induced an increase in the barrier function of the 2D HTM monolayers. A one-day exposure to a 2 mg/L solution of ATX also resulted in a significant decrease in the sizes of the 3D HTM spheroids, and an increase in stiffness was also observed. The gene expression of several ECMs, their regulators and ER-stress related factors by the 3D HTM spheroids were altered by both ATX and LPA, but in different manners. Conclusions: The findings presented herein suggest that ATX may have additional roles in the human TM, in addition to the ATX–LPA signaling axis.

Fatty Liver Index(FLI)は新規高血圧発症の独立した予測因子である

東浦幸村, 古橋眞人, 田中希尚, 高橋聖子, 山下智久, 小山雅之, 大西浩文, 斎藤重幸, 三浦哲嗣, 塙なぎさ

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 43回 217 - 217 2021.10

一般住民におけるFatty Liver Index(FLI)と高血圧発症との関連

東浦幸村, 古橋眞人, 田中希尚, 大西浩文, 三浦哲嗣

日本動脈硬化学会総会プログラム・抄録集 ( (一社)日本動脈硬化学会 ) 53回 186 - 186 2021.10

Fatty Liver Index(FLI)は新規高血圧発症の独立した予測因子である

東浦幸村, 古橋眞人, 田中希尚, 高橋聖子, 山下智久, 小山雅之, 大西浩文, 斎藤重幸, 三浦哲嗣, 塙なぎさ

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 43回 217 - 217 2021.10

一般住民におけるFatty Liver Index(FLI)と高血圧発症との関連

東浦幸村, 古橋眞人, 田中希尚, 大西浩文, 三浦哲嗣

日本動脈硬化学会総会プログラム・抄録集 ( (一社)日本動脈硬化学会 ) 53回 186 - 186 2021.10

抗リン脂質抗体症候群の腎機能障害への関与が示唆されたループス腎炎の1例

赤澤史子, 田中希尚, 山下智久, 寺沢誠, 長南新太, 高橋聖子, 小松弘明, 安部功記, 矢野俊之, 古橋眞人, 土橋和文, 小川弥生

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 63 ( 6-E ) 676 - 676 2021.09

Distinct Regulation of U-ACE2 and P-ACE2 (Urinary and Plasma Angiotensin-Converting Enzyme 2) in a Japanese General Population

Masato Furuhashi, Akiko Sakai, Marenao Tanaka, Yukimura Higashiura, Kazuma Mori, Masayuki Koyama, Hirofumi Ohnishi, Shigeyuki Saitoh, Kazuaki Shimamoto

Hypertension ( Ovid Technologies (Wolters Kluwer Health) ) 2021.08

Severe acute respiratory syndrome coronavirus 2 in coronavirus disease 2019 invades the host through ACE (angiotensin-converting enzyme) 2 as the host cellular receptor for a viral spike protein. ACE2 converts angiotensin II to angiotensin-(1–7) and cleaved ACE2 is detectable in urine and plasma. However, regulation of U-ACE2 (urinary ACE2) and P-ACE2 (plasma ACE2) and their alterations by renin-angiotensin-aldosterone system inhibitors remain unclear. We simultaneously investigated U-ACE2 and P-ACE2 in 605 Japanese participants (male/female: 280/325, mean age: 65±15 years) in the Tanno-Sobetsu cohort study in 2017. Males had significantly lower U-ACE2 and higher P-ACE2 than did females. There was no significant correlation between U-ACE2 and P-ACE2. P-ACE2 was significantly lower in subjects treated with renin-angiotensin-aldosterone system inhibitors than in those not treated with renin-angiotensin-aldosterone system inhibitors, but there was no significant difference in U-ACE2 between the groups. Multivariable regression analyses showed that female sex, high levels of systolic blood pressure, hemoglobin A1c, and urinary albumin-to-creatinine ratio, and low uric acid level were independent predictors of high U-ACE2 level and that high levels of γ-glutamyl transpeptidase, estimated glomerular filtration rate, and uric acid were independent predictors of high P-ACE2 level. In conclusion, U-ACE2 and P-ACE2 are distinctly regulated and the use of renin-angiotensin-aldosterone system inhibitors is not an independent predictor of their levels in a Japanese general population. U-ACE2 is associated with high blood pressure, high glucose level, and microalbuminuria, and low urate level, whereas P-ACE2 is associated with liver dysfunction, high glomerular filtration rate, and high urate level.

Independent Association of Fatty Liver Index With Left Ventricular Diastolic Dysfunction in Subjects Without Medication

Masato Furuhashi, Atsuko Muranaka, Satoshi Yuda, Marenao Tanaka, Masayuki Koyama, Mina Kawamukai-Nishida, Satoko Takahashi, Yukimura Higashiura, Daisuke Miyamori, Ryo Nishikawa, Hirofumi Ohnishi, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

The American Journal of Cardiology ( Elsevier BV ) 2021.08

Elevated Fatty Liver Index Is Independently Associated With New Onset of Hypertension During a 10‐Year Period in Both Male and Female Subjects

Yukimura Higashiura, Masato Furuhashi, Marenao Tanaka, Satoko Takahashi, Kazuma Mori, Daisuke Miyamori, Masayuki Koyama, Hirofumi Ohnishi, Norihito Moniwa, Keita Numata, Takashi Hisasue, Nagisa Hanawa, Tetsuji Miura

Journal of the American Heart Association ( Ovid Technologies (Wolters Kluwer Health) ) 10 ( 14 ) 2021.07

Background Fatty liver index (FLI), a predictor of nonalcoholic fatty liver disease, has been reported to be associated with several metabolic disorders. Because of a sex difference in FLI level, we hypothesized that FLI is associated with development of hypertension to a greater extent in men or women. Methods and Results We investigated the relationship between FLI and development of hypertension during a 10‐year period in a general population of subjects who received annual health examinations (n=28 990). After exclusion (44.9%) of subjects with missing data and those with hypertension at baseline, a total of 15 965 subjects (men/women: 9466/6499) were included. FLI level was significantly higher in men than in women. During the 10‐year period, 2304 men (24.3%) and 745 women (11.5%) had new onset of hypertension. Multivariable Cox proportional hazard models with a restricted cubic spline showed that the hazard ratios (HRs) for development of hypertension after adjustment of age, systolic blood pressure, estimated glomerular filtration rate, habits of smoking and alcohol drinking, family history of hypertension, and diagnosis of diabetes mellitus and dyslipidemia increased gradually with increase in FLI in men and increased rapidly and then slowly with increase in FLI in women. There was a significant interaction between FLI and sex for the risk of hypertension in all of the subjects ( P =0.049). The addition of FLI to traditional risk factors significantly improved the discriminatory capability. Conclusions A high level of FLI predicts the development of hypertension in both men and women, although distribution patterns of HRs were different between sexes.

Seasonal variation of serum 25-hydroxyvitamin D level in hemodialysis patients in the northernmost island of Japan

Yufu Gocho, Marenao Tanaka, Hirohito Sugawara, Masato Furuhashi, Norihito Moniwa, Tomohisa Yamashita, Hideki Takizawa, Hiroya Mukai, Kouhei Ohno, Takuto Maeda, Arata Osanami, Hirofumi Ohnishi, Hiroaki Komatsu, Kazuma Mori, Tetsuji Miura

Clinical and Experimental Nephrology ( Springer Science and Business Media {LLC} ) 2021.07

Screening of the Drug-Induced Effects of Prostaglandin EP2 and FP Agonists on 3D Cultures of Dexamethasone-Treated Human Trabecular Meshwork Cells

Megumi Watanabe, Yosuke Ida, Masato Furuhashi, Yuri Tsugeno, Hiroshi Ohguro, Fumihito Hikage

Biomedicines 2021.07

Involvement of necroptosis in contrast-induced nephropathy in a rat CKD model

Satoru Shibata, Norihito Moniwa, Atsushi Kuno, Ayumu Kimura, Wataru Ohwada, Hirohito Sugawara, Yufu Gocho, Marenao Tanaka, Toshiyuki Yano, Masato Furuhashi, Masaya Tanno, Takayuki Miki, Tetsuji Miura

Clinical and Experimental Nephrology ( Springer Science and Business Media {LLC} ) 25 ( 7 ) 708 - 717 2021.07 [Domestic journal]

BACKGROUND: The risk of contrast-induced nephropathy (CIN) is high in patients with chronic kidney disease (CKD). However, the mechanism of CIN in CKD is not fully understood. Here, we prepared a clinically relevant model of CIN and examined the role of necroptosis, which potentially cross-talks with autophagy, in CIN. METHODS: In Sprague-Dawley rats, CKD was induced by subtotal nephrectomy (SNx, 5/6 nephrectomy) 4 weeks before induction of CIN. CIN was induced by administration of a contrast medium (CM), iohexol, following administration of indomethacin and N-omega-Nitro-L-arginine methyl ester. Renal function and tissue injuries were assessed 48 h after CM injection. RESULTS: Serum creatinine (s-Cre) and BUN were increased from 0.28 ± 0.01 to 0.52 ± 0.02 mg/dl and from 15.1 ± 0.7 to 29.2 ± 1.2 mg/dl, respectively, after SNx alone. CM further increased s-Cre and BUN to 0.69 ± 0.03 and 37.2 ± 2.1, respectively. In the renal tissue after CM injection, protein levels of receptor-interacting serine/threonine-protein kinase (RIP) 1, RIP3, cleaved caspase 3, and caspase 8 were increased by 64 ~ 212%, while there was reduction in LC3-II and accumulation of p62. Necrostatin-1, an RIP1 inhibitor, administered before and 24 h after CM injection significantly suppressed elevation of s-Cre, BUN and urinary albumin levels, kidney injury molecule-1 expression and infiltration of CD68-positive macrophages in renal tissues after CM injection. CONCLUSION: The results suggest that necroptosis of proximal tubular cells contributes to CIN in CKD and that suppression of protective autophagy by pro-necroptotic signaling may also be involved.

Independent association of plasma xanthine oxidoreductase activity with hypertension in nondiabetic subjects not using medication

Masato Furuhashi, Yukimura Higashiura, Masayuki Koyama, Marenao Tanaka, Takayo Murase, Takashi Nakamura, Seigo Akari, Akiko Sakai, Kazuma Mori, Hirofumi Ohnishi, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

Hypertension Research ( Springer Science and Business Media {LLC} ) 44 ( 9 ) 1213 - 1220 2021.06 [International journal]

Xanthine oxidoreductase (XOR), a rate-limiting and catalyzing enzyme of uric acid formation in purine metabolism, is involved in reactive oxygen species generation. Plasma XOR activity has been shown to be a novel metabolic biomarker related to obesity, liver dysfunction, hyperuricemia, dyslipidemia, and insulin resistance. However, the association between plasma XOR activity and hypertension has not been fully elucidated. We investigated the association of hypertension with plasma XOR activity in 271 nondiabetic subjects (male/female: 119/152) who had not taken any medications in the Tanno-Sobetsu Study, a population-based cohort. Males had higher plasma XOR activity than females. Plasma XOR activity was positively correlated with mean arterial pressure (r = 0.128, P = 0.036). When the subjects were divided by the presence and absence of hypertension into an HT group (male/female: 34/40) and a non-HT group (male/female: 85/112), plasma XOR activity in the HT group was significantly higher than that in the non-HT group (median: 39 vs. 28 pmol/h/mL, P = 0.028). There was no significant difference in uric acid levels between the two groups. Multivariable logistic regression analysis showed that plasma XOR activity (odds ratio: 1.091 [95% confidence interval: 1.023-1.177] per 10 pmol/h/mL, P = 0.007) was an independent determinant of the risk for hypertension after adjustment for age, sex, current smoking and alcohol consumption, estimated glomerular filtration rate, brain natriuretic peptide, and insulin resistance index. The interaction of sex with plasma XOR activity was not significant for the risk of hypertension. In conclusion, plasma XOR activity is independently associated with hypertension in nondiabetic individuals who are not taking any medications.

CKDモデルの造影剤腎症におけるネクロプトーシスの役割

柴田智, 茂庭仁人, 木村歩, 後町結, 田中希尚, 古橋眞人, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 63 ( 4 ) 447 - 447 2021.06

CKD発症予測におけるfatty liver indexの意義

高橋聖子, 田中希尚, 古橋眞人, 長南新太, 宮森大輔, 後町結, 塙なぎさ, 茂庭仁人, 大西浩文, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 63 ( 4 ) 453 - 453 2021.06

血液透析患者におけるビタミンDの季節変動に関する検討

田中希尚, 後町結, 菅原浩仁, 茂庭仁人, 山下智久, 古橋眞人, 滝沢英毅, 向博也, 大野紘平, 三浦哲嗣

日本透析医学会雑誌 ( (一社)日本透析医学会 ) 54 ( Suppl.1 ) 379 - 379 2021.05

Antiatherosclerotic Phenotype of Perivascular Adipose Tissue Surrounding the Saphenous Vein in Coronary Artery Bypass Grafting

Takuma Mikami, Masato Furuhashi, Akiko Sakai, Ryosuke Numaguchi, Ryo Harada, Syuichi Naraoka, Takeshi Kamada, Yukimura Higashiura, Marenao Tanaka, Shunsuke Ohori, Taku Sakurada, Masanori Nakamura, Yutaka Iba, Joji Fukada, Tetsuji Miura, Nobuyoshi Kawaharada

Journal of the American Heart Association ( Ovid Technologies (Wolters Kluwer Health) ) 10 ( 7 ) OJ32 - 2 2021.04

Background Perivascular adipose tissue (PVAT) is associated with metabolically driven chronic inflammation called metaflammation, which contributes to vascular function and the pathogenesis of vascular disease. The saphenous vein (SV) is commonly used as an essential conduit in coronary artery bypass grafting, but the long‐term patency of SV grafts is a crucial issue. The use of the novel “no‐touch” technique of SV harvesting together with its surrounding tissue has been reported to result in good long‑term graft patency of SV grafts. Herein, we investigated whether PVAT surrounding the SV (SV‐PVAT) has distinct phenotypes compared with other PVATs of vessels. Methods and Results Fat pads were sampled from 48 patients (male/female, 32/16; age, 72±8 years) with coronary artery disease who underwent elective coronary artery bypass grafting. Adipocyte size in SV‐PVAT was significantly larger than the sizes in PVATs surrounding the internal thoracic artery, coronary artery, and aorta. SV‐PVAT and PVAT surrounding the internal thoracic artery had smaller extents of fibrosis, decreased gene expression levels of fibrosis‐related markers, and less metaflammation, as indicated by a significantly smaller extent of cluster of differentiation 11c–positive M1 macrophage infiltration, higher gene expression level of adiponectin, and lower gene expression levels of inflammatory cytokines, than did PVATs surrounding the coronary artery and aorta. Expression patterns of adipocyte developmental and pattern‐forming genes were totally different among the PVATs of the vessels. Conclusions The phenotype of SV‐PVAT, which may result from inherent differences in adipocytes, is closer to that of PVAT surrounding the internal thoracic artery than that of PVAT surrounding the coronary artery or that of PVAT surrounding the aorta. SV‐PVAT has less metaflammation and consecutive adipose tissue remodeling, which may contribute to high long‐term patency of grafting when the no‐touch technique of SV harvesting is used.

Impact of atrial fibrillation on the risk of ischemic stroke in patients on hemodialysis: BOREAS-HD3 Study

Ayumu Kimura, Marenao Tanaka, Norihito Moniwa, Arata Osanami, Koki Abe, Daisuke Miyamori, Yufu Gocho, Satoru Shibata, Makoto Terasawa, Yusuke Okazaki, Tomohisa Yamashita, Masayuki Koyama, Masato Furuhashi, Hirofumi Ohnishi, Tetsuji Miura

Clinical and Experimental Nephrology ( Springer Science and Business Media {LLC} ) 25 ( 3 ) 297 - 304 2021.03

血中FABP4濃度の上昇からみた一般集団における心血管死の予測 12年間の前向き研究(Elevated Circulating FABP4 Concentration Predicts Cardiovascular Death in a General Population: A 12-year Prospective Study)

齋藤礼衣, 古橋眞人, 小山雅之, 東浦幸村, 赤坂憲, 田中希尚, 茂庭仁人, 大西浩文, 齋藤重幸, 浦信行, 島本和明, 三浦哲嗣

日本循環器学会学術集会抄録集 ( (一社)日本循環器学会 ) 85回 OJ16 - 4 2021.03

Fatty acid-binding protein 4 is an independent factor in the pathogenesis of retinal vein occlusion

Fumihito Hikage, Masato Furuhashi, Yosuke Ida, Hiroshi Ohguro, Megumi Watanabe, Soma Suzuki, Kaku Itoh

PLOS ONE ( Public Library of Science ({PLoS}) ) 16 ( 1 ) 2021.01

The main objective of current study was to identify the fatty acid-binding protein 4 (FABP4) expressed in both adipocytes and macrophages in vitreous fluid from patients with retinal vein occlusion (RVO). Patients with RVO (n = 14, CRVO; central RVO n = 5, BRVO; branch RVO n = 9) and non-RVO (macular hole or epiretinal membrane, n = 18) were surgically treated by a 25 or 27G vitrectomy. Undiluted vitreous fluid samples obtained as the result of surgery were subjected to enzyme-linked immunosorbent assays to measure the levels of FABP4 and vascular endothelial growth factor A (VEGFA).Data including ocular blood flow by laser speckle flow graphy (LSFG), height and weight, systemic blood pressures and several blood biochemistry values were collected. Among the LSFG mean blur rate (MBR) values of the optic nerve head (ONH) at baseline, MA (MBR of all area), MV (MBR of the vascular area), and MV-MT (MBR of the tissue area) were significantly decreased in patients with CRVO. The levels of V-FABP4 and V-VEGFA were relatively or significantly (P< 0.05) higher in the BRVO or CRVO patients compared to the non-RVO patients, respectively. A positive correlation (r = 0.36, P = 0.045) or a negative correlation (r = -0.51, P = 0.006) was observed between Log V-FABP4 and Log V-VEGF, or Log V-FABP4 and MV-MT at post-operative 1-week, respectively. Furthermore, neither of these factors were affected with respect to sex, body mass index and several clinical parameters that were collected, except that a positive correlation was observed for Log V-FABP4 with blood urea nitrogen. Stepwise multivariable regression analyses indicated that MV-MT at post-operative 1week was independently associated with Log V-FABP4 after adjustment for age and gender, and gender and Log V-FABP4 were independently associated with Log V-VEGFA after adjustment for age. The findings reported herein suggest that an independent factor, FABP4 may be synergistically involved in the pathogenesis of RVO with VEGFA.

Prediction of new onset of diabetes mellitus during a 10-year period by using a combination of levels of alanine aminotransferase and γ-glutamyl transferase

Daisuke Miyamori, Marenao Tanaka, Masato Furuhashi, Hirofumi Ohnishi, Masayuki Koyama, Arata Osanami, Yukimura Higashiura, Keita Numata, Takashi Hisasue, Nagisa Hanawa, Norihito Moniwa, Tetsuji Miura

Endocrine Journal ( Japan Endocrine Society ) 2021

Prediction and validation of nonalcoholic fatty liver disease by fatty liver index in a Japanese population

Satoko Takahashi, Marenao Tanaka, Yukimura Higashiura, Kazuma Mori, Nagisa Hanawa, Hirofumi Ohnishi, Masato Furuhashi

Endocrine Journal ( Japan Endocrine Society ) 2021

Circulating PCSK7 Level is Independently Associated with Obesity, Triglycerides Level and Fatty Liver Index in a General Population without Medication

Masato Furuhashi, Yu Kataoka, Ryo Nishikawa, Masayuki Koyama, Akiko Sakai, Yukimura Higashiura, Marenao Tanaka, Shigeyuki Saitoh, Kazuaki Shimamoto, Hirofumi Ohnishi

Journal of Atherosclerosis and Thrombosis ( Japan Atherosclerosis Society ) 2021

A Resuscitated Case of Acute Myocardial Infarction with both Familial Hypercholesterolemia Phenotype Caused by Possibly Oligogenic Variants of the <i>PCSK9</i> and <i>ABCG5</i> Genes and Type I CD36 Deficiency

Ryo Nishikawa, Masato Furuhashi, Mika Hori, Masatsune Ogura, Mariko Harada-Shiba, Takeshi Okada, Masahiro Koseki, Takeshi Kujiraoka, Hiroaki Hattori, Ryosuke Ito, Atsuko Muranaka, Nobuaki Kokubu, Tetsuji Miura

Journal of Atherosclerosis and Thrombosis ( Japan Atherosclerosis Society ) 2021 [Domestic journal]

A 56-year-old postmenopausal woman with out-of-hospital cardiac arrest caused by acute myocardial infraction was successfully resuscitated by intensive treatments and recovered without any neurological disability. She was diagnosed as having familial hypercholesterolemia (FH) based on a markedly elevated low-density lipoprotein cholesterol (LDL-C) level and family history of premature coronary artery disease. Genetic testing in her family members showed that a variant of the proprotein convertase subtilisin/kexin type 9 (PCSK9) gene (c.2004C＞A, p.S668R), which had been previously reported as having uncertain significance, was associated with FH, indicating that the variant is a potential candidate for the FH phenotype. Next-generation sequencing analysis for the proband also showed that there was a heterozygous mutation of the ATP-binding cassette sub-family G member 5 (ABCG5) gene (c.1166G＞A, R389H), which has been reported to increase LDL-C level and the risk of cardiovascular disease. She was also diagnosed as having type 1 CD36 deficiency based on a lack of myocardial uptake of 123 I-labeled 15-(p-iodophenyl)-3-R,S-methyl-pentadecanoic acid in scintigraphy and the absence of CD36 antigen in both monocytes and platelets in flow cytometry. She had a homozygous mutation of the CD36 gene (c.1126-5_1127delTTTAGAT), which occurs in a canonical splice site (acceptor) and is predicted to disrupt or distort the normal gene product. To our knowledge, this is the first report of a heterozygous FH phenotype caused by possibly oligogenic variants of the PCSK9 and ABCG5 genes complicated with type I CD36 deficiency caused by a novel homozygous mutation. Both FH phenotype and CD36 deficiency might have caused extensive atherosclerosis, leading to acute myocardial infarction in the present case.

Significance of urinary fatty acid-binding protein 4 level as a possible biomarker for the identification of minimal change disease in patents with nephrotic-range proteinuria

Marenao Tanaka, Masato Furuhashi, Norihito Moniwa, Takuto Maeda, Hideki Takizawa, Megumi Matsumoto, Akiko Sakai, Yukimura Higashiura, Yufu Gocho, Masayuki Koyama, Yayoi Ogawa, Tetsuji Miura

BMC Nephrology ( Springer Science and Business Media {LLC} ) 21 ( 1 ) 2020.12

Abstract Background Fatty acid-binding protein 4 (FABP4), but not FABP1 (liver-type FABP), is ectopically induced in injured glomerular endothelial cells, and urinary FABP4 (U-FABP4) level is associated with proteinuria and renal dysfunction in a general population. Methods The clinical significance of U-FABP4 was investigated in 81 patients (male/female: 43/38, age: 57 ± 17 years) who underwent kidney biopsy. Results U-FABP4 was negatively correlated with estimated glomerular filtration rate (eGFR) (r = − 0.56, P < 0.01) and was positively correlated with age, blood pressure, triglycerides, proteinuria (r = 0.58, P < 0.01), plasma FABP4 and urinary FABP1 (U-FABP1) (r = 0.52, P < 0.01). Multivariable regression analysis showed that eGFR, proteinuria and U-FABP1 were independent predictors of U-FABP4. The level of U-FABP4, but not that of proteinuria, eGFR or U-FABP1, in minimal change nephrotic syndrome (MCNS) was significantly lower than the level in membranous nephropathy (MN) and that in diabetic nephropathy. Receiver operating characteristic curve analysis indicated that U-FABP4 level ≤ 0.78 μg/gCr predicted MCNS in patients who had nephrotic-range proteinuria with a high level of accuracy. When divided by the median value of U-FABP4 at baseline in 33 of the 81 patients who could be followed up, the yearly change (post–pre) in eGFR in the low U-FABP4 group was significantly greater than that in the high U-FABP4 group (median: 11.0 vs. -5.0 mL/min/1.73m2/year). Conclusions U-FABP4 level is independently associated with proteinuria and renal dysfunction in patients with glomerular kidney disease. A low U-FABP4 level may predict MCNS in patients with nephrotic syndrome and would be a useful biomarker for differential diagnosis of MCNS and MN, which are common causes of nephrotic syndrome.

Treatment with anagliptin, a DPP-4 inhibitor, decreases FABP4 concentration in patients with type 2 diabetes mellitus at a high risk for cardiovascular disease who are receiving statin therapy

Masato Furuhashi, Ichiro Sakuma, Takeshi Morimoto, Yukimura Higashiura, Akiko Sakai, Megumi Matsumoto, Mio Sakuma, Michio Shimabukuro, Takashi Nomiyama, Osamu Arasaki, Koichi Node, Shinichiro Ueda

Cardiovascular Diabetology ( Springer Science and Business Media {LLC} ) 19 ( 1 ) 2020.12

Abstract Background Fatty acid-binding protein 4 (FABP4) acts as a novel adipokine, and elevated FABP4 concentration is associated with obesity, insulin resistance and atherosclerosis. Dipeptidyl peptidase-4 (DPP-4) inhibitors, a class of antidiabetic drugs, have distinct structures among the drugs, possibly leading to a drug class effect and each drug effect. Sitagliptin, a DPP-4 inhibitor, has been reported to decrease FABP4 concentration in drug-naïve and sulfonylurea-treated patients with type 2 diabetes mellitus. Anagliptin, another DPP-4 inhibitor, was shown to decrease low-density lipoprotein cholesterol (LDL-C) level to a greater extent than that by sitagliptin in the Randomized Evaluation of Anagliptin vs. Sitagliptin On low-density lipoproteiN cholesterol in diabetes (REASON) trial. Aim and methods As a sub-analysis study using data obtained from the REASON trial, we investigated the effects of treatment with anagliptin (n = 148, male/female: 89/59) and treatment with sitagliptin (n = 159, male/female: 93/66) for 52 weeks on FABP4 concentration in patients with type 2 diabetes mellitus at a high risk for cardiovascular events who were receiving statin therapy. Results The DPP-4 inhibitor had been administered in 82% of the patients in the anagliptin group and 81% of the patients in sitagliptin group prior to randomization. Serum FABP4 level was significantly decreased by 7.9% by treatment with anagliptin (P = 0.049) and was not significantly decreased by treatment with sitagliptin (P = 0.660). Change in FABP4 level was independently associated with basal FABP4 level and changes in waist circumference and creatinine after adjustment of age, sex and the treatment group. Conclusion Anagliptin decreases serum FABP4 concentration independent of change in hemoglobin A1c or LDL-C in patients with type 2 diabetes mellitus and dyslipidemia who are on statin therapy. Trial registration ClinicalTrials.gov number NCT02330406. Registered January 5, 2015, https://clinicaltrials.gov/ct2/show/NCT02330406

New insights into purine metabolism in metabolic diseases: role of xanthine oxidoreductase activity

Masato Furuhashi

American Journal of Physiology-Endocrinology and Metabolism ( American Physiological Society ) 319 ( 5 ) E827 - E834 2020.11

Xanthine oxidoreductase (XOR) consists of two different forms, xanthine dehydrogenase and xanthine oxidase (XO), and is a rate-limiting enzyme of uric acid production from hypoxanthine and xanthine. Uric acid is the end product of purine metabolism in humans and has a powerful antioxidant effect. The lack of ascorbic acid, known as vitamin C, in hominoids has been thought to cause a compensatory increase in uric acid as an antioxidant by unfunctional gene mutation of uricase to a pseudogene. Because XO is involved in an increase in reactive oxygen species (ROS) by generating superoxide and hydrogen peroxide, inadequate activation of XOR promotes oxidative stress-related tissue injury. Plasma XOR activity is associated with obesity, smoking, liver dysfunction, hyperuricemia, dyslipidemia, insulin resistance, and adipokines, indicating a novel biomarker of metabolic disorders. However, XOR activity in adipose tissue is low in humans unlike in rodents, and hypoxanthine is secreted from human adipose tissue. The concentration of hypoxanthine, but not xanthine, is independently associated with obesity in a general population, indicating differential regulation of hypoxanthine and xanthine. Treatment with an XOR inhibitor can decrease uric acid for preventing gout, reduce production of XO-related ROS, and promote reutilization of hypoxanthine and ATP production through the salvage pathway. It has recently been suggested that discontinuation of an XOR inhibitor causes adverse cardiovascular outcomes as XOR inhibitor withdrawal syndrome, possibly due to cardiac disturbance of conduction and contraction by reduced ATP production. New insights into purine metabolism, including the role of XOR activity in the past 5 yr, are mainly discussed in this review.

Independent and Distinct Associations of FABP4 and FABP5 With Metabolic Parameters in Type 2 Diabetes Mellitus

Masato Furuhashi, Ichiro Sakuma, Takeshi Morimoto, Yukimura Higashiura, Akiko Sakai, Megumi Matsumoto, Mio Sakuma, Michio Shimabukuro, Takashi Nomiyama, Osamu Arasaki, Koichi Node, Shinichiro Ueda

Frontiers in Endocrinology ( Frontiers Media {SA} ) 11 575557 - 575557 2020.09 [International journal]

Objective: Among fatty acid-binding proteins (FABPs), secreted forms of FABP4 and FABP5, which are expressed in adipocytes and macrophages, act as bioactive molecules. We investigated concentrations of FABP4 and FABP5 in patients with type 2 diabetes mellitus. Methods: As a sub-analysis study of the Randomized Evaluation of Anagliptin vs. Sitagliptin On low-density lipoproteiN cholesterol in diabetes (REASON) trial, 256 patients (male/female: 146/110, age: 68 ± 10 years) with type 2 diabetes mellitus and dyslipidemia who were receiving statin therapy were recruited. Patients who had been treated with a thiazolidinedione were excluded. Results: Several drugs which may modulate FABP4 levels including statins, dipeptidyl peptidase-4 inhibitors and angiotensin II receptor blockers had been administered in 100, 81, and 51% of the recruited patients, respectively. The level of FABP4, but not that of FABP5, was significantly higher in females than in males. Multivariable linear regression analysis demonstrated that waist circumference (β = 0.21), estimated glomerular filtration rate (β = -0.31), triglycerides (β = 0.16), and FABP5 (β = 0.39) were independent predictors of FABP4 level after adjusting age and sex. On the other hand, FABP5 level was independently associated with levels of FABP4 (β = 0.57) and high-density lipoprotein (HDL) cholesterol (β = -0.12). Conclusions: Concentrations of FABP4 and FABP5 are independent predictors of each other in patients with type 2 diabetes mellitus. There are distinct independent associations of FABP4 with renal dysfunction, adiposity and hypertriglyceridemia and there is a distinct independent association of FABP5 with a low HDL cholesterol level in type 2 diabetic patients with dyslipidemia at high risks for cardiovascular disease who are receiving statin therapy.

U-shaped relationship between serum uric acid level and decline in renal function during a 10-year period in female subjects: BOREAS-CKD2.

Kazuma Mori, Masato Furuhashi, Marenao Tanaka, Keita Numata, Takashi Hisasue, Nagisa Hanawa, Masayuki Koyama, Arata Osanami, Yukimura Higashiura, Masafumi Inyaku, Megumi Matsumoto, Norihito Moniwa, Hirofumi Ohnishi, Tetsuji Miura

Hypertension research : official journal of the Japanese Society of Hypertension ( Springer Science and Business Media {LLC} ) 44 ( 1 ) 107 - 116 2020.08 [Refereed] [International journal]

While hyperuricemia is recognized as a risk factor for chronic kidney disease (CKD), the risk of CKD in subjects with a low level of serum uric acid (UA) remains controversial. Here, we examined whether the association of CKD risk with serum UA level differs depending on the sex and age of subjects in a general population. Of subjects who received annual health checkups, we enrolled 6,779 subjects (male/female: 4,454/2,325; age: 45 ± 9 years) with data from a 10-year follow-up after excluding subjects taking anti-hyperuricemic drugs and those with CKD at baseline. During the follow-up period, 11.4% of the males and 11.7% of the females developed CKD. A significant interaction of sex, but not age, with the effect of baseline UA level on CKD risk was found. A restricted cubic spline analysis showed a U-shaped association of the baseline UA level with the risk of CKD in females. Multivariable Cox proportional hazard analyses for females showed that baseline UA levels in the 5th quintile (Q5, ≥5 mg/dL; HR: 1.68) and the 1st quintile (Q1, ≤3.5 mg/dL; HR: 1.73) were independent risk factors for CKD when compared with UA levels in the 4th quintile (Q4, 4.5-4.9 mg/dL). In males, restricted cubic spline analysis indicated increased CKD risk in subjects with a higher baseline UA level but not in those with a low UA level. In conclusion, a low UA level is a significant risk factor for CKD in females, while an elevated UA level increases the risk of CKD in both sexes.

Potential differential effects of renin-angiotensin system inhibitors on SARS-CoV-2 infection and lung injury in COVID-19.

Masato Furuhashi, Norihito Moniwa, Hideki Takizawa, Nobuyuki Ura, Kazuaki Shimamoto

Hypertension research : official journal of the Japanese Society of Hypertension 2020.05 [Refereed] [International journal]

糖尿病外来診療における慢性合併症評価の実態調査

小川俊史, 佐藤達也, 東浦幸村, 伊野祥哉, 矢野俊之, 古橋眞人, 丹野雅也, 三木隆幸, 斎藤重幸, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 63 ( 5 ) 356 - 356 2020.05

Low urine pH predicts new onset of diabetes mellitus during a 10-year period in men: BOREAS-DM1 study.

Yukimura Higashiura, Marenao Tanaka, Masato Furuhashi, Masayuki Koyama, Hirofumi Ohnishi, Keita Numata, Takashi Hisasue, Nagisa Hanawa, Norihito Moniwa, Tetsuji Miura

Journal of diabetes investigation 11 ( 6 ) 1490 - 1497 2020.04 [Refereed] [Domestic journal]

AIMS/INTRODUCTION: A low level of urine pH (U-pH) has been reported to be associated with metabolic disorders. However, the relationship between the incidence of diabetes mellitus and U-pH has not yet been fully addressed. MATERIALS AND METHODS: We investigated the relationship between U-pH and the development of diabetes mellitus during a 10-year period in a general population of individuals who received annual health examinations in 2006 (n = 28,990). After exclusion of individuals with missing data, and those with diabetes mellitus and/or chronic kidney disease at baseline, a total of 12,476 individuals (men/women: 8,027/4,449) who received health examinations at least once during the period from 2007 to 2016 were recruited. The recruited individuals were divided into four groups according to their U-pH levels: groups of U-pH ≤5.0, 5.5, 6.0 and ≥6.5. RESULTS: During a 10-year period, 521 men (6.5%) and 132 women (3.0%) had new onset of diabetes mellitus. The cumulative incidence of diabetes mellitus was 7.5% (men/women: 9.3%/4.4%) per 100 person-years. The hazard ratios (HRs) in the U-pH ≤5.0 (HR 1.93) and U-pH 5.5 groups (HR 1.46) were significantly higher than that in the U-pH ≥6.5 group as a reference for men, but not for women. After adjustment of age, obesity, fasting glucose, smoking and alcohol drinking habits, family history of diabetes mellitus, and use of drugs for hypertension and dyslipidemia, HR in the U-pH ≤5.0 group (HR 1.39) was significantly higher than that in the U-pH ≥6.5 group for men, but not for women. CONCLUSIONS: Low U-pH predicts new onset of diabetes mellitus in a general population of men.

ALT、γGTPの組み合わせによる糖尿病発症リスク評価の向上

宮森大輔, 田中希尚, 古橋眞人, 茂庭仁人, 三浦哲嗣, 小山雅之, 大西浩文, 沼田啓太, 久末章史, 塙なぎさ

日本内科学会雑誌 ( (一社)日本内科学会 ) 109 ( Suppl. ) 241 - 241 2020.02

ALT、γGTPの組み合わせによる糖尿病発症リスク評価の向上

宮森大輔, 田中希尚, 古橋眞人, 茂庭仁人, 三浦哲嗣, 小山雅之, 大西浩文, 沼田啓太, 久末章史, 塙なぎさ

日本内科学会雑誌 ( (一社)日本内科学会 ) 109 ( Suppl. ) 241 - 241 2020.02

Differential regulation of hypoxanthine and xanthine by obesity in a general population.

Masato Furuhashi, Masayuki Koyama, Yukimura Higashiura, Takayo Murase, Takashi Nakamura, Megumi Matsumoto, Akiko Sakai, Hirofumi Ohnishi, Marenao Tanaka, Shigeyuki Saitoh, Norihito Moniwa, Kazuaki Shimamoto, Tetsuji Miura

Journal of diabetes investigation 11 ( 4 ) 878 - 887 2020.01 [Refereed] [Domestic journal]

AIMS/INTRODUCTION: Uric acid is synthesized by oxidation of hypoxanthine and xanthine using a catalyzing enzyme, xanthine oxidoreductase (XOR), which can be a source of reactive oxygen species. Plasma XOR activity is a metabolic biomarker associated with obesity, hyperuricemia, liver dysfunction and insulin resistance. However, it has recently been reported that XOR activity in fat tissue is low in humans, unlike in rodents, and that hypoxanthine is secreted from human fat tissue. MATERIALS AND METHODS: The associations of obesity with hypoxanthine, xanthine and plasma XOR activity were investigated in 484 participants (men/women: 224/260) of the Tanno-Sobetsu Study. RESULTS: Levels of hypoxanthine, xanthine and plasma XOR activity were significantly higher in men than in women. In 59 participants with hyperuricemia, 11 (men/women: 11/0) participants were being treated with an XOR inhibitor and had a significantly higher level of xanthine, but not hypoxanthine, than that in participants without treatment. In all of the participants, hypoxanthine concentration in smokers was significantly higher than that in non-smokers. Stepwise and multivariate regression analyses showed that body mass index, smoking habit and xanthine were independent predictors of hypoxanthine after adjustment of age, sex and use of antihyperuricemic drugs. Whereas, alanine transaminase, hypoxanthine and plasma XOR activity were independent predictors for xanthine, and alanine transaminase, triglycerides and xanthine were independent predictors for plasma XOR activity. CONCLUSIONS: The concentration of hypoxanthine, but not that of xanthine, is independently associated with obesity and smoking habit, indicating differential regulation of hypoxanthine and xanthine in a general population.

高intact PTH血症、高Ca血症から副甲状腺癌の遠隔再発の診断を得た一例

寺沢誠, 後町結, 小山雅之, 茂庭仁人, 古橋眞人, 三浦哲嗣

日本内分泌学会雑誌 ( (一社)日本内分泌学会 ) 95 ( 3 ) 864 - 864 2020.01

Reply to the comment of Hirota et al. on "Accuracy of flash glucose monitoring in insulin-treated patients with type 2 diabetes".

Tatsuya Sato, Hiroto Oshima, Kei Nakata, Yukishige Kimura, Toshiyuki Yano, Masato Furuhashi, Masaya Tannno, Takayuki Miki, Tetsuji Miura

Journal of diabetes investigation 11 ( 1 ) 256 - 256 2020.01 [Domestic journal]

一般住民においてヒポキサンチンとキサンチンの血中濃度は肥満によって異なった制御を受ける

古橋眞人, 東浦幸村, 小山雅之, 村瀬貴代, 中村敬志, 酒井晶子, 田中希尚, 茂庭仁人, 大西浩文, 斎藤重幸, 三浦哲嗣

日本痛風・核酸代謝学会総会プログラム抄録集 ( (一社)日本痛風・尿酸核酸学会 ) 53回 74 - 74 2020.01

SAP療法下に多量飲酒を誘因に発症した糖尿病性ケトアシドーシスの1例

東浦幸村, 中田圭, 西川諒, 矢野俊之, 古橋眞人, 三木隆幸, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 62 ( 12 ) 776 - 776 2019.12

北海道都市部健診受診者コホートにおける初年度の血圧カテゴリ別の血清尿酸値と高血圧発症との関連

小山雅之, 樋室伸顕, 田中希尚, 茂庭仁人, 古橋眞人, 沼田啓太, 久末章史, 塙なぎさ, 齋藤重幸, 三浦哲嗣, 大西浩文

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 42回 300 - 300 2019.10

一般住民において血漿ヒポキサンチン濃度は肥満度および喫煙と関連する

東浦幸村, 古橋眞人, 松本めぐみ, 酒井晶子, 小山雅之, 村瀬貴代, 中村敬志, 田中希尚, 茂庭仁人, 大西浩文, 斎藤重幸, 三浦哲嗣

肥満研究 ( (一社)日本肥満学会 ) 25 ( Suppl. ) 279 - 279 2019.10

一般住民におけるキサンチン酸化還元酵素(XOR)活性と高血圧との関連

東浦幸村, 古橋眞人, 小山雅之, 田中希尚, 茂庭仁人, 松本めぐみ, 酒井晶子, 村瀬貴代, 中村敬志, 大西浩文, 齋藤重幸, 島本和明, 三浦哲嗣

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 42回 279 - 279 2019.10

Annual change in plasma xanthine oxidoreductase activity is associated with changes in liver enzymes and body weight.

Masato Furuhashi, Masayuki Koyama, Megumi Matsumoto, Takayo Murase, Takashi Nakamura, Yukimura Higashiura, Marenao Tanaka, Norihito Moniwa, Hirofumi Ohnishi, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

Endocrine journal 66 ( 9 ) 777 - 786 2019.09 [Refereed] [Domestic journal]

Xanthine oxidoreductase (XOR), an enzyme of uric acid formation from hypoxanthine and xanthine, is recognized as a source of oxidative stress. Plasma activity of XOR has been reported to be a biomarker of metabolic disorders associated with obesity, liver dysfunction, insulin resistance, hyperuricemia and adipokines. We investigated longitudinal change in plasma XOR activity, which was determined by using mass spectrometry and liquid chromatography to detect [13C2, 15N2]-uric acid using [13C2, 15N2]-xanthine as a substrate, in 511 subjects (male/female: 244/267) of the Tanno-Sobetsu Study in the years 2016 and 2017. Plasma XOR activity in a basal state was significantly higher in men than in women, but no significant sex difference was observed in annual change in plasma XOR activity. Annual change in plasma activity of XOR was positively correlated with changes in each parameter, including body weight (r = 0.203, p < 0.001), body mass index, diastolic blood pressure, aspartate transaminase (AST) (r = 0.772, p < 0.001), alanine transaminase (r = 0.647, p < 0.001), γ-glutamyl transpeptidase, total cholesterol, triglycerides, uric acid, fasting glucose and HbA1c. Multivariate regression analysis demonstrated that change in AST and that in body weight were independent predictors of change in plasma XOR activity after adjustment of age, sex and changes in each variable with a significant correlation without multicollinearity. In conclusion, annual change in plasma XOR activity is independently associated with changes in liver enzymes and body weight in a general population. Improvement of liver function and reduction of body weight would decrease plasma XOR activity and its related oxidative stress as a therapeutic strategy.

Anorexia nervosaによるCKDの3症例

後町結, 茂庭仁人, 木村歩, 長南新太, 菅原浩仁, 高橋聖子, 田中希尚, 古橋眞人, 小川弥生, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 61 ( 6 ) 739 - 739 2019.08

ネフローゼ症候群を呈したシェーグレン症候群による二次性膜性腎症に対しステロイドが奏効した1例

増田拳, 田中希尚, 茂庭仁人, 後町結, 古橋眞人, 小川弥生, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 61 ( 6 ) 762 - 762 2019.08

ゴリムマブが誘因となった薬剤起因性自己免疫疾患でネフローゼ症候群を発症した1例

長谷川諒, 安部功記, 松久優雅, 小林広学, 田中希尚, 後町結, 茂庭仁人, 古橋眞人, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 61 ( 6 ) 700 - 700 2019.08

潜在性甲状腺機能亢進症による二次性膜性腎症と診断した1例

茂庭仁人, 大嶋優, 後町結, 高橋聖子, 田中希尚, 古橋眞人, 黒田せつ子, 浜武継, 島友子, 小川弥生, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 61 ( 6 ) 699 - 699 2019.08

Independent links between plasma xanthine oxidoreductase activity and levels of adipokines.

Masato Furuhashi, Megumi Matsumoto, Takayo Murase, Takashi Nakamura, Yukimura Higashiura, Masayuki Koyama, Marenao Tanaka, Norihito Moniwa, Hirofumi Ohnishi, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

Journal of diabetes investigation 10 ( 4 ) 1059 - 1067 2019.07 [Refereed] [Domestic journal]

AIMS/INTRODUCTION: Xanthine oxidoreductase (XOR) is a rate-limiting enzyme that catalyzes uric acid formation in the purine metabolism, is involved in an increase in reactive oxygen species. Plasma XOR activity has been shown to be associated with obesity, smoking, liver dysfunction, hyperuricemia, dyslipidemia and insulin resistance. MATERIALS AND METHODS: The association between plasma XOR activity, measured by using liquid chromatography and mass spectrometry, and levels of adipokines, including adiponectin, fatty acid-binding protein 4 (FABP4) and fibroblast growth factor 21 (FGF21), was investigated in 282 participants (male/female: 126/156) of the Tanno-Sobetsu Study who were not taking medication. RESULTS: Women had lower plasma XOR activity than did men. Smoking habit was associated with increased activity. Plasma XOR activity was positively correlated with concentrations of FABP4 (r = 0.192, P < 0.001) and FGF21 (r = 0.208, P < 0.001), homeostasis model assessment of insulin resistance as an index of insulin resistance and uric acid, and was negatively correlated with adiponectin level (r = -0.243, P = 0.001). Multivariate regression analyses showed that levels of adiponectin, FABP4 and FGF21 were independent determinants of plasma XOR activity after adjusting age, sex, uric acid and homeostasis model assessment of insulin resistance. With additional adjustment of smoking habit, the level of FABP4, but not that of adiponectin or FGF21, remained as an independent predictor of plasma XOR activity. CONCLUSIONS: Plasma XOR activity was independently associated with levels of adipokines in a general population of individuals not taking medication.

心肺停止を伴う急性心筋梗塞を契機に診断に至った1型CD36欠損症および家族性高コレステロール血症の合併例

西川諒, 古橋眞人, 伊藤良介, 續太郎, 小山雅之, 村中敦子, 國分宣明, 堀美香, 小倉正恒, 斯波真理子, 三浦哲嗣

日本動脈硬化学会総会プログラム・抄録集 ( (一社)日本動脈硬化学会 ) 51回 1 - 3 2019.07

Accuracy of flash glucose monitoring in insulin-treated patients with type 2 diabetes.

Tatsuya Sato, Hiroto Oshima, Kei Nakata, Yukishige Kimura, Toshiyuki Yano, Masato Furuhashi, Masaya Tanno, Takayuki Miki, Tetsuji Miura

Journal of diabetes investigation 10 ( 3 ) 846 - 850 2019.05 [Domestic journal]

The present study evaluated the accuracy of interstitial glucose measurements by flash glucose monitoring (FGM) and continuous glucose monitoring (CGM). Five diabetes patients simultaneously underwent FGM (FreeStyle Libre Pro) and CGM (iPro™2), and their glucose levels were compared with venous blood and capillary blood glucose levels. The range of daily venous blood glucose levels (30 measurements) was 70-245 mg/dL, with a median of 138 mg/dL. There were good correlations of glucose levels measured by FGM (r2 = 0.90, mean absolute relative difference 8.2 ± 5.6%), CGM (r2 = 0.86, mean absolute relative difference 9.2 ± 9.1%) and capillary blood (r2 = 0.87, mean absolute relative difference 7.2 ± 7.2%) with venous blood glucose levels. The accuracy of FGM measurements was also shown against CGM, with 99.9% of the FGM values (1,279 measurements) being within the Parkes error grid zones A and B. The results suggest that the accuracy of FGM is similar to that of CGM, and that FGM is a useful tool for determining daily glucose profile.

糖尿病の外来診療における慢性合併症評価実態の検討

伊野祥哉, 中田圭, 東浦幸村, 佐藤達也, 矢野俊之, 古橋眞人, 丹野雅也, 三木隆幸, 斎藤重幸, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 62 ( Suppl.1 ) S - 306 2019.04

Differential Phenotypes in Perivascular Adipose Tissue Surrounding the Internal Thoracic Artery and Diseased Coronary Artery.

Ryosuke Numaguchi, Masato Furuhashi, Megumi Matsumoto, Hiroshi Sato, Yosuke Yanase, Yosuke Kuroda, Ryo Harada, Toshiro Ito, Yukimura Higashiura, Masayuki Koyama, Marenao Tanaka, Norihito Moniwa, Masanori Nakamura, Hirosato Doi, Tetsuji Miura, Nobuyoshi Kawaharada

Journal of the American Heart Association 8 ( 2 ) e011147 2019.01 [Refereed] [International journal]

Background Perivascular adipose tissue ( PVAT ) is causally associated with vascular function and the pathogenesis of vascular disease in association with metabolically driven chronic inflammation called metaflammation. However, the difference in PVAT surrounding the coronary artery ( CA - PVAT ) and that surrounding the internal thoracic artery (ITA-PVAT), a vessel resistant to atherosclerosis, remains unclear. Herein, we investigated whether CA - PVAT , ITA - PVAT , and subcutaneous adipose tissue ( SCAT ) have distinct phenotypes. Methods and Results Fat pads were sampled from 44 patients (men/women, 36:8; age, 67±13 years) with CA disease who underwent elective CA bypass grafting. Adipocyte size in ITA - PVAT and that in CA - PVAT were significantly smaller than that in SCAT . A greater extent of fibrosis and increased gene expression levels of fibrosis-related molecules were observed in CA - PVAT than those in SCAT and those in ITA - PVAT . CA - PVAT exhibited more pronounced metaflammation, as indicated by a significantly larger extent of CD 68-positive and CD 11c-positive M1 macrophages, a lower ratio of CD 206-positive M2 to CD 11c-positive M1 macrophages, a lower gene expression level of adiponectin, and higher gene expression levels of inflammatory cytokines and inflammasome- and endoplasmic reticulum stress-related molecules, than did ITA - PVAT and SCAT . Expression patterns of adipocyte developmental and pattern-forming genes were totally different among SCAT , ITA - PVAT, and CA - PVAT . Conclusions The phenotype of ITA - PVAT is closer to that of SCAT than that of CA - PVAT , which may result from inherent differences in adipocytes. ITA - PVAT appears to be protected from metaflammation and consecutive adipose tissue remodeling, which may contribute to the decreased atherosclerotic plaque burden in the ITA.

Unexpected high plasma xanthine oxidoreductase activity in female subjects with low levels of uric acid.

Masato Furuhashi, Kazuma Mori, Marenao Tanaka, Takuto Maeda, Megumi Matsumoto, Takayo Murase, Takashi Nakamura, Masayuki Koyama, Norihito Moniwa, Hirofumi Ohnishi, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

Endocrine journal 65 ( 11 ) 1083 - 1092 2018.11 [Refereed] [Domestic journal]

Hypouricemia is a high-risk factor of exercise-induced acute kidney injury (EIAKI) probably through a lack of an antioxidant effect of uric acid. Xanthine oxidoreductase (XOR) is an enzyme that catalyzes the formation of uric acid from hypoxanthine and xanthine, leading to an increase in superoxide and reactive oxygen species. Activation of XOR has been proposed to promote oxidative stress-related tissue injury. We measured plasma XOR activity by a sensitive and accurate assay using a combination of liquid chromatography and triple quadrupole mass spectrometry in subjects with relatively low levels of uric acid (≤4.0 mg/dL) who were recruited from 627 subjects (male/female: 292/335) in the Tanno-Sobetsu Study, a population-based cohort. The numbers of subjects with uric acid ≤4.0 mg/dL, ≤3.0 mg/dL and ≤2.0 mg/dL were 72 (11.5%, male/female: 5/67), 13 (2.1%, all females) and 2 (0.3%, both females), respectively. Plasma XOR activities in 5 male subjects were below the median value of the 292 male subjects. In 12 (17.9%) of the 67 female subjects with uric acid ≤4.0 mg/dL, plasma XOR activities were above the upper quartile value of the 335 female subjects. Eleven of the 12 female subjects with high plasma XOR activity and a low uric acid level had liver dysfunction and/or insulin resistance. In conclusion, unexpected high plasma XOR activities were found in some female subjects with relatively low levels of uric acid. Measurement of plasma XOR activity may help to identify hypouricemic patients with a high risk for EIAKI.

【腎と高血圧:最新研究から得られた新たな知見】CKDの新規治療ターゲットとしての異所性FABP4発現の可能性

古橋眞人, 東浦幸村, 田中希尚

血圧 ( (株)先端医学社 ) 25 ( 9 ) 630 - 635 2018.09

脂肪酸結合蛋白4(FABP4/A-FABP/aP2)は、主に脂肪細胞とマクロファージに存在し、メタボリックシンドロームの成因に深く関わる。FABP4はシグナルペプチドを有さないものの脂肪細胞から脂肪分解とともに分泌されてアディポカインとして作用する。また、血中FABP4濃度は肥満、糖尿病、脂質異常症、高血圧、動脈硬化、心血管死と関連する。現在、尿中FABP1(L-FABP)排泄量が腎尿細管機能障害マーカーとして臨床応用されているが、我々は傷害された糸球体内皮細胞にFABP1ではなくFABP4が異所性に発現することを初めて見出し、糸球体FABP4発現量や尿中FABP4排泄量が尿蛋白量や腎予後に関わることを報告した。CKDのうち特に糸球体傷害疾患に対して、異所性FABP4発現が新規の治療ターゲットとなる可能性が示唆される。(著者抄録)

糖尿病合併IgA腎症に対し扁摘パルス療法を施行した3症例

茂庭仁人, 後町結, 田中希尚, 古橋眞人, 小川弥生, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 60 ( 6 ) 937 - 937 2018.08

Plasma Xanthine Oxidoreductase Activity as a Novel Biomarker of Metabolic Disorders in a General Population.

Masato Furuhashi, Megumi Matsumoto, Marenao Tanaka, Norihito Moniwa, Takayo Murase, Takashi Nakamura, Hirofumi Ohnishi, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

Circulation journal : official journal of the Japanese Circulation Society 82 ( 7 ) 1892 - 1899 2018.06 [Refereed] [Domestic journal]

BACKGROUND: Xanthine oxidoreductase (XOR) is an enzyme that catalyzes the formation of uric acid from hypoxanthine and xanthine, leading to an increase in superoxide and reactive oxygen species. Activation of XOR promotes oxidative stress-related tissue injury. We investigated the associations between metabolic parameters and plasma XOR activity measured by a sensitive and accurate assay using a combination of liquid chromatography and triple quadrupole mass spectrometry to detect [13C2,15N2]-uric acid using [13C2,15N2]-xanthine as a substrate.Methods and Results:A total of 627 Japanese subjects (M/F, 292/335) from the Tanno-Sobetsu Study, a population-based cohort, were recruited. Plasma XOR activity was significantly higher in males than in females, and habitual smoking was associated with elevation of activity. Plasma XOR activity was positively correlated with body mass index (BMI; r=0.323, P<0.001), waist circumference, blood pressure, and levels of liver enzymes including alanine transaminase (r=0.694, P<0.001), uric acid (r=0.249, P<0.001), triglycerides (r=0.312, P<0.001), hemoglobin A1c, fasting glucose, insulin and HOMA-R (r=0.238, P<0.001) as a marker of insulin resistance and was negatively correlated with high-density lipoprotein cholesterol level. On stepwise and multivariate regression analyses, BMI, smoking and levels of alanine transaminase, uric acid, triglycerides and HOMA-R were independent predictors of plasma XOR activity after adjustment for age and gender. CONCLUSIONS: Plasma XOR activity is a novel biomarker of metabolic disorders in a general population.

良性小脳髄膜腫切除後に寛解に至った膜性腎症の1例

森和真, 山下智久, 安部功記, 木村幸滋, 田中希尚, 茂庭仁人, 古橋眞人, 小川弥生, 三浦哲嗣

日本内科学会雑誌 ( (一社)日本内科学会 ) 107 ( 6 ) 1102 - 1106 2018.06

症例は69歳女性で、嘔気を伴う頭痛を自覚し、救急搬送された。意識障害に加え、CTで右小脳に石灰化を伴う4cm大の腫瘍とそれに伴う水頭症を認めた。頭部MRI T2強調で、腫瘍は不均一な高信号を示した。第2病日に緊急脳室ドレナージ、頭蓋内腫瘍摘出術を施行し、摘出病理標本から良性線維性髄膜腫と診断された。入院時から尿蛋白(4+)、血清Albは2.5→1.3g/dlまで低下し、第6病日から4日間アルブミン製剤を投与した。腫瘍切除後に蛋白尿は速やかに減少した。全身状態が改善した第28病日に腎生検を施行し、膜性腎症と診断した。さらに、特発性膜性腎症に特異的とされる抗PLA2R抗体染色陽性、IgGサブクラス染色でのIgG4優位を認めた。電子顕微鏡所見で上皮下に高電子密度沈着物(EDD)を認め、一部の電子密度は低下していることから、Ehrenreich Churg分類Stage III〜IVと診断した。第35病日に尿蛋白量は24時間蓄尿検査で0.4g/日まで減少した。髄膜腫の抗PLA2R抗体染色を行うも陰性であった。

脂肪酸結合タンパク4(FABP4)濃度は動脈硬化進展度の予測因子である

古橋眞人, 湯田聡, 村中敦子, 川向美奈, 松本めぐみ, 田中希尚, 茂庭仁人, 大西浩文, 斎藤重幸, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 61 ( Suppl.1 ) S - 319 2018.04

急性心不全入院における急性腎障害予測スコア(S-ARS)は1年後の死亡を予測する

柴田智, 山下智久, 菅原浩仁, 田中希尚, 茂庭仁人, 古橋眞人, 三木隆幸, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 60 ( 3 ) 357 - 357 2018.04

腎疾患症例における尿FABP4排泄量の臨床的意義腎生検例を用いた検討

田中希尚, 古橋眞人, 松本めぐみ, 山下智久, 茂庭仁人, 後町結, 前田卓人, 滝沢英毅, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 60 ( 3 ) 423 - 423 2018.04

一般住民における尿酸低値例での血漿キサンチン酸化還元酵素(XOR)活性の検討

森和真, 古橋眞人, 田中希尚, 村瀬貴代, 中村敬志, 前田卓人, 大西浩文, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 60 ( 3 ) 365 - 365 2018.04

Flash Glucose Monitoringを用いた血糖値予測妥当性の検討

青山ちひろ, 三木隆幸, 大島洸人, 中田圭, 佐藤達也, 矢野俊之, 古橋眞人, 齋藤重幸, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 61 ( Suppl.1 ) S - 191 2018.04

Circulating Fatty Acid-Binding Protein 4 Concentration Predicts the Progression of Carotid Atherosclerosis in a General Population Without Medication.

Masato Furuhashi, Satoshi Yuda, Atsuko Muranaka, Mina Kawamukai, Megumi Matsumoto, Marenao Tanaka, Norihito Moniwa, Hirofumi Ohnishi, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

Circulation journal : official journal of the Japanese Circulation Society 82 ( 4 ) 1121 - 1129 2018.03 [Refereed] [Domestic journal]

BACKGROUND: Fatty acid-binding protein 4 (FABP4), which is expressed in both adipocytes and macrophages, is secreted from the cells and acts as an adipokine. An elevated circulating FABP4 level is associated with insulin resistance and atherosclerosis.Methods and Results:We investigated the causative association between FABP4 level and progression of atherosclerosis in subjects of the Tanno-Sobetsu Study, a population-based cohort. In 281 subjects without medication (male/female: 109/172) in the year 2010 or 2013, the carotid intima-media thickness (CIMT) assessed using carotid ultrasonography was significantly correlated with age, adiposity, blood pressure, renal dysfunction and levels of cholesterol, triglycerides, fasting glucose, HbA1c and FABP4 (r=0.331, P<0.001). Multiple regression analysis demonstrated that age, sex and FABP4 concentration were independent predictors of CIMT. A total of 78 (male/female: 29/49) of the 156 subjects in 2010 underwent carotid ultrasonography again in 2013. The change in CIMT each year during that 3-year period (mean±SD: 3.8±22.3 µm/year) was positively correlated with basal levels of high-sensitivity C-reactive protein (hsCRP) (r=0.231, P=0.046) and FABP4 (r=0.267, P=0.018) in 2010. After adjustment for age, sex and hsCRP level, the basal FABP4 level was independently associated with the change in CIMT per year. CONCLUSIONS: FABP4 concentration is an independent predictor of the progression of carotid atherosclerosis.

糖尿病や検尿異常のない場合もLDL-コレステロール高値はCKDの危険因子となる

長南新太, 山下智久, 松本環, 大西浩文, 塙なぎさ, 田中希尚, 茂庭仁人, 古橋眞人, 三浦哲嗣

日本内科学会雑誌 ( (一社)日本内科学会 ) 107 ( Suppl. ) 252 - 252 2018.02

A Case of Crescentic Glomerulonephritis Complicated with Hypocomplementemic Urticarial Vasculitis Syndrome and ANCA-Associated Vasculitis

Marenao Tanaka, Norihito Moniwa, Tomohiro Mita, Toshiyuki Tobisawa, Tamaki Matsumoto, Atsushi Mochizuki, Tomohisa Yamashita, Toshiyuki Yano, Masato Furuhashi, Tetsuji Miura

Case Reports in Nephrology and Dialysis ( S. Karger AG ) 7 ( 3 ) 144 - 153 2017.11 [Refereed]

慢性腎臓病研究の進歩から見えてきた高血圧成因慢性腎臓病の新規治療ターゲットとしての異所性FABP4発現の可能性

古橋眞人, 田中希尚, 三浦哲嗣

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 40回 255 - 255 2017.10

Ectopic Fatty Acid-Binding Protein 4 Expression in the Vascular Endothelium is Involved in Neointima Formation After Vascular Injury

Takahiro Fuseya, Masato Furuhashi, Megumi Matsumoto, Yuki Watanabe, Kyoko Hoshina, Tomohiro Mita, Shutaro Ishimura, Marenao Tanaka, Tetsuji Miura

JOURNAL OF THE AMERICAN HEART ASSOCIATION ( WILEY ) 6 ( 9 ) 2017.09 [Refereed]

Background-Fatty acid-binding protein 4 (FABP4) is expressed in adipocytes, macrophages, and endothelial cells of capillaries but not arteries. FABP4 is secreted from adipocytes in association with lipolysis, and an elevated circulating FABP4 level is associated with obesity, insulin resistance, and atherosclerosis. However, little is known about the link between FABP4 and endovascular injury. We investigated the involvement of ectopic FABP4 expression in endothelial cells in neointima hyperplasia after vascular injury. Methods and Results-Femoral arteries of 8-week-old male mice were subjected to wire-induced vascular injury. After 4 weeks, immunofluorescence staining showed that FABP4 was ectopically expressed in endothelial cells of the hyperplastic neointima. Neointima formation determined by intima area and intima to media ratio was significantly decreased in FABP4-defficient mice compared with that in wild-type mice. Adenovirus-mediated overexpression of FABP4 in human coronary artery endothelial cells (HCAECs) in vitro increased inflammatory cytokines and decreased phosphorylation of nitric oxide synthase 3. Furthermore, FABP4 was secreted from HCAECs. Treatment of human coronary smooth muscle cells or HCAECs with the conditioned medium of Fabp4-overexpressed HCAECs or recombinant FABP4 significantly increased gene expression of inflammatory cytokines and proliferationand adhesion-related molecules in cells, promoted cell proliferation and migration of human coronary smooth muscle cells, and decreased phosphorylation of nitric oxide synthase 3 in HCAECs, which were attenuated in the presence of an anti-FABP4 antibody. Conclusions-Ectopic expression and secretion of FABP4 in vascular endothelial cells contribute to neointima formation after vascular injury. Suppression of ectopic FABP4 in the vascular endothelium would be a novel strategy against post-angioplasty vascular restenosis.

ベバシズマブにより糸球体内皮障害と高度蛋白尿を呈し薬剤中止とARBにて軽快した一例

木村幸滋, 山下智久, 田中希尚, 茂庭仁人, 古橋眞人, 小川弥生, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 59 ( 6 ) 915 - 915 2017.09

高度腎機能障害で発症したが自然軽快したTINU症候群の一例

小川俊史, 山下智久, 田中希尚, 茂庭仁人, 古橋眞人, 小川弥生, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 59 ( 6 ) 895 - 895 2017.09

本態性血小板血症に続発した軽鎖沈着症による末期腎不全の一例

安部功記, 茂庭仁人, 山下智久, 田中希尚, 古橋眞人, 村瀬和幸, 池田博, 杉田真太朗, 黒田せつ子, 小川弥生, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 59 ( 6 ) 893 - 893 2017.09

良性小脳髄膜腫による水頭症を合併し、術後に完全寛解に至った膜性腎症の一例

森和真, 山下智久, 安部功記, 木村幸滋, 田中希尚, 茂庭仁人, 古橋眞人, 小川弥生, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 59 ( 6 ) 882 - 882 2017.09

肺高血圧症に伴うCKDの特徴

茂庭仁人, 山下智久, 田中希尚, 古橋眞人, 永野伸卓, 西田絢一, 村中敦子, 國分宣明, 藤戸健史, 望月敦史, 永原大五, 橋本暁佳, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 59 ( 3 ) 362 - 362 2017.04

非糖尿病かつ検尿異常のない場合もLDL-コレステロール120mg/dL以上はCKD-ステージG3への進展の危険因子となる

山下智久, 松本環, 大西浩文, 塙なぎさ, 田中希尚, 茂庭仁人, 古橋眞人, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 59 ( 3 ) 372 - 372 2017.04

SGLT2阻害薬カナグリフロジン投与による脂肪酸結合タンパク4(FABP4)濃度の経時的変化とその規定因子の検討

古橋眞人, 松本めぐみ, 平光伸也, 田中希尚, 茂庭仁人, 石井潤一, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 60 ( Suppl.1 ) S - 361 2017.04

急性心不全における急性腎障害予測スコアにフロセミド静注量は必要か?

柴田智, 山下智久, 田中希尚, 茂庭仁人, 古橋眞人, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 59 ( 3 ) 374 - 374 2017.04

多腺性自己免疫症候群3A型に房室ブロックを合併した一例抗Ro/SS-A抗体との関連

中田圭, 矢野俊之, 大島洸人, 古橋眞人, 丹野雅也, 斎藤重幸, 三木隆幸, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 60 ( Suppl.1 ) S - 452 2017.04

Serum FABP5 concentration is a potential biomarker for residual risk of atherosclerosis in relation to cholesterol efflux from macrophages

Masato Furuhashi, Masatsune Ogura, Megumi Matsumoto, Satoshi Yuda, Atsuko Muranaka, Mina Kawamukai, Akina Omori, Marenao Tanaka, Norihito Moniwa, Hirofumi Ohnishi, Shigeyuki Saitoh, Mariko Harada-Shiba, Kazuaki Shimamoto, Tetsuji Miura

SCIENTIFIC REPORTS ( NATURE PUBLISHING GROUP ) 7 ( 1 ) 217 2017.03 [Refereed]

Cholesterol efflux capacity (CEC) from macrophages, the first step in the reverse cholesterol transport pathway, is inversely associated with residual risk for atherosclerotic cardiovascular disease. Fatty acid-binding protein 4 (FABP4) and FABP5 are expressed in both adipocytes and macrophages and play significant roles in the development of insulin resistance and atherosclerosis. Both FABP4 and FABP5 are secreted from cells, and their circulating levels are associated with insulin resistance and atherosclerosis. We investigated the association between CEC and levels of FABP4 and FABP5 in 250 subjects without any medications. CEC was positively correlated with HDL cholesterol level and negatively correlated with concentrations of high-sensitivity C-reactive protein (hsCRP) and FABP5, but not FABP4. Multiple regression analysis demonstrated that FABP5 concentration was an independent predictor of CEC after adjustment of age, gender and levels of HDL cholesterol and hsCRP. In 129 of the 250 subjects who underwent carotid ultrasonography, mean intima-media thickness was negatively correlated with CEC and was positively correlated with concentrations of FABP4 and FABP5. In conclusion, in contrast to FABP4, circulating FABP5 is associated with decreased CEC and carotid atherosclerosis, suggesting that FABP5 level is a regulatory factor of CEC and a potential biomarker for residual risk of atherosclerosis.

Impact of the Number of Anti-Thrombosis Agents in Hemodialysis Patients: BOREAS-HD2 Study

Marenao Tanaka, Norihito Moniwa, Hirofumi Ohnishi, Tomohisa Yamashita, Masayuki Koyama, Yufu Gocho, Keitaro Nishizawa, Yukishige Kimura, Hirohito Sugawara, Sayaka Murakami, Yusuke Okazaki, Masato Furuhashi, Hideaki Yoshida, Tetsuji Miura

KIDNEY & BLOOD PRESSURE RESEARCH ( KARGER ) 42 ( 3 ) 553 - 564 2017 [Refereed]

Background/Aims: Relationships between the number of anti-thrombosis agents, clinical benefits and adverse events in hemodialysis (HD) patients are unclear. Methods: All patients on HD in 22 institutes (n = 1,071) were enrolled and followed up for 3 years. After exclusion of patients with missing data, kidney transplantation or retraction of consent during the follow-up period (n = 204), mortality rate and ischemic and hemorrhagic events were compared between different regimens of anti-thrombosis agents. Results: The use of dual or triple antiplatelet (AP) agents (HR:2.03, 95% CI:1.01-4.13, p = 0.04) and the combination of an AP agent and warfarin (WF) (HR:4.84, 95% CI 1.96-11.96, p < 0.001) were associated with an increase in hemorrhagic events compared with no use of anti-thrombosis agents. No anti-thrombosis regimen was associated with a significant change in risk of ischemic stroke. The use of dual or triple AP agents, but not WF, was associated with an increase in cardiovascular mortality (HR:2.48, 95% CI:1.24-4.76, p = 0.01). Conclusion: A significant increase in hemorrhagic events by the use of dual or more AP agents and by co-administration of an AP agent and WF in patients on HD should be considered in planning their anti-thrombosis regimen. (C) 2017 The Author(s) Published by S. Karger AG, Basel

Transcriptome and Metabolome Analyses in Exogenous FABP4-and FABP5-Treated Adipose-Derived Stem Cells

Tokunori Yamamoto, Masato Furuhashi, Takeshi Sugaya, Tsuyoshi Oikawa, Megumi Matsumoto, Yasuhito Funahashi, Yoshihisa Matsukawa, Momokazu Gotoh, Tetsuji Miura

PLOS ONE ( PUBLIC LIBRARY SCIENCE ) 11 ( 12 ) e0167825 2016.12 [Refereed]

Adipose-derived stem cells (ADSC), which exist near adipocytes in adipose tissue, have been used as a potential tool of regenerative medicine. Lipid chaperones, fatty acid-binding protein 4 (FABP4) and 5 (FABP5), are abundantly expressed in adipocytes. FABP4 has recently been shown to be secreted from adipocytes during lipolysis in a non-classical pathway and may act as an adipokine. Here, we investigated the role of exogenous FABP4 and FABP5 in transcriptional and metabolic regulation in ADSC. FABP4 and FABP5 were little expressed in ADSC. However, both FABP4 and FABP5 were significantly induced after adipocyte differentiation of ADSC and were secreted from the differentiated adipocytes. Analysis of microarray data, including gene ontology enrichment analysis and cascade analysis of the protein-protein interaction network using a transcription factor binding site search, demonstrated that treatment of ADSC with FABP4 or FABP5 affected several kinds of genes related to inflammatory and metabolic responses and the process of cell differentiation. Notably, myogenic factors, including myocyte enhancer factors, myogenic differentiation 1 and myogenin, were modulated by treatment of ADSC with FABP4, indicating that exogenous FABP4 treatment is partially associated with myogenesis in ADSC. Metabolome analysis showed that treatment of ADSC with FABP4 and with FABP5 similarly, but differently in extent, promoted hydrolysis and/or uptake of lipids, consequentially together with enhancement of beta oxidation, inhibition of downstream of the glycolysis pathway, accumulation of amino acids, reduction of nucleic acid components and increase in the ratio of reduced and oxidized nicotinamide adenine dinucleotide phosphates (NADPH/NADP(+)), an indicator of reducing power, and the ratio of adenosine triphosphate and adenosine monophosphate (ATP/AMP), an indicator of the energy state, in ADSC. In conclusion, secreted FABP4 and FABP5 from adipocytes as adipokines differentially affect transcriptional and metabolic regulation in ADSC near adipocytes. The adiposity condition in the host of regenerative medicine may affect characteristics of ADSC by exposure of the balance of FABP4 and FABP5.

糖尿病性腎症による末期腎不全に至ってから診断されたCushing病の1例

高橋遼, 舘越勇輝, 茂庭仁人, 西田絢一, 矢野俊之, 古橋眞人, 三木隆幸, 三浦哲嗣

日本内分泌学会雑誌 ( (一社)日本内分泌学会 ) 92 ( S.Branc ) 13 - 13 2016.12

2型糖尿病患者においてipragliflozinは他のSGLT2阻害薬と比較し降圧効果に優れる

茂庭仁人, 山下智久, 田中希尚, 古橋眞人, 三浦哲嗣

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 39回 350 - 350 2016.09

造影剤腎症、薬剤性急性間質性腎炎との鑑別に難渋したコレステロール塞栓症の一例

西沢慶太郎, 山下智久, 田中希尚, 茂庭仁人, 古橋眞人, 野口寛子, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 58 ( 6 ) 820 - 820 2016.08

Transplantation associated-thrombotic microangiopathy(TA-TMA)の二例

安部功記, 山下智久, 田中希尚, 茂庭仁人, 古橋眞人, 小川弥生, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 58 ( 6 ) 767 - 767 2016.08

Independent Link Between Levels of Proprotein Convertase Subtilisin/Kexin Type 9 and FABP4 in a General Population Without Medication.

Masato Furuhashi, Akina Omori, Megumi Matsumoto, Yu Kataoka, Marenao Tanaka, Norihito Moniwa, Hirofumi Ohnishi, Hideaki Yoshida, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

The American journal of cardiology 118 ( 2 ) 198 - 203 2016.07 [Refereed] [International journal]

Proprotein convertase subtilisin/kexin type 9 (PCSK9) binds to and degrades the low-density lipoprotein (LDL) receptor, leading to hypercholesterolemia and cardiovascular risk. Fatty acid binding protein 4 (FABP4/adipocyte FABP/aP2) is secreted from adipocytes in association with lipolysis, and circulating FABP4 has been reported to act as an adipokine for the development of insulin resistance and atherosclerosis. Elevated serum FABP4 level is associated with obesity, insulin resistance, dyslipidemia, and atherosclerosis. In this study, we examined the association between circulating levels of FABP4 and PCSK9 in a general population. A total of 265 subjects (male/female: 98/167) who were not on medication were recruited from subjects of the Tanno-Sobetsu Study, and concentrations of FABP4 and PCSK9 were measured. The level of FABP4, but not that of PCSK9, showed a gender difference, being higher in women than in men. FABP4 level was independently associated with gender, adiposity, renal dysfunction, and levels of cholesterol and PCSK9. There was a significant and gender-different correlation between PCSK9 level and age: negatively in men (r = -0.250, p = 0.013) and positively in women (r = 0.183, p = 0.018). After adjustment of age, gender, and LDL cholesterol level, PCSK9 level was positively and independently correlated with FABP4 concentration. In conclusion, PCSK9 level is differentially regulated by gender during aging. Circulating FABP4 is independently associated with the PCSK9 level, suggesting that elevation of FABP4 level as an adipokine leads to dyslipidemia through increased PCSK9 level and subsequent degradation of the LDL receptor.

Impact of use of angiotensin II receptor blocker on all-cause mortality in hemodialysis patients: prospective cohort study using a propensity-score analysis.

Marenao Tanaka, Tomohisa Yamashita, Masayuki Koyama, Norihito Moniwa, Kohei Ohno, Kaneto Mitsumata, Takahito Itoh, Masato Furuhashi, Hirofumi Ohnishi, Hideaki Yoshida, Kazufumi Tsuchihashi, Tetsuji Miura

Clinical and experimental nephrology 20 ( 3 ) 469 - 78 2016.06 [Refereed] [Domestic journal]

BACKGROUND: It is controversial whether treatment with an angiotensin II receptor blocker (ARB) or a calcium channel blocker (CCB) improves prognosis of hemodialysis (HD) patients. METHODS: This study was designed as a multicenter prospective cohort study. HD patients (n = 1071) were enrolled from 22 institutes in January 2009 and followed up for 3 years. Patients with missing data, kidney transplantation or retraction of consent during the follow-up period (n = 204) were excluded, and 867 patients contributed to analysis of mortality. Propensity score (PS) for use of ARB and that for CCB was calculated using a multiple logistic regression model. RESULTS: ARB and CCB were prescribed in 45.6 and 54.7 % of patients at enrollment. During the 3-year follow-up period, all-cause mortality and cardiovascular mortality rates were 18.8 and 5.1 %, respectively. Kaplan-Meier curves showed that all-cause and cardiovascular mortality rates were lower in the ARB group than in the non-ARB group, though the mortality rates were similar in the CCB group and non-CCB group. In PS-stratified Cox regression analysis, ARB treatment was associated with 34 and 45 % reduction of all-cause death and cardiovascular death, respectively. In PS matching analysis, ARB treatment was associated with a significant reduction (46 % reduction) in the risk of all-cause death. A significant impact of CCB treatment on all-cause or cardiovascular mortality was not detected in PS analysis. CONCLUSIONS: The use of an ARB, but not a CCB, is associated with reduced all-cause and cardiovascular mortalities in patients on HD.

家族性高コレステロール血症における残余リスクマーカーとしての脂肪酸結合蛋白

小倉正恒, 古橋眞人, 森本めぐむ, 松本めぐみ, 三浦哲嗣, 斯波真理子

日本動脈硬化学会総会プログラム・抄録集 ( (一社)日本動脈硬化学会 ) 48回 201 - 201 2016.06

Local Production of Fatty Acid-Binding Protein 4 in Epicardial/Perivascular Fat and Macrophages Is Linked to Coronary Atherosclerosis

Masato Furuhashi, Takahiro Fuseya, Masaki Murata, Kyoko Hoshina, Shutaro Ishimura, Tomohiro Mita, Yuki Watanabe, Akina Omori, Megumi Matsumoto, Takeshi Sugaya, Tsuyoshi Oikawa, Junichi Nishida, Nobuaki Kokubu, Marenao Tanaka, Norihito Moniwa, Hideaki Yoshida, Norimasa Sawada, Kazuaki Shimamoto, Tetsuji Miura

ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY ( LIPPINCOTT WILLIAMS & WILKINS ) 36 ( 5 ) 825 - 834 2016.05 [Refereed]

Objective-Fatty acid-binding protein 4 (FABP4) is expressed in adipocytes and macrophages, and elevated circulating FABP4 level is associated with obesity-mediated metabolic phenotype. We systematically investigated roles of FABP4 in the development of coronary artery atherosclerosis. Approach and Results-First, by immunohistochemical analyses, we found that FABP4 was expressed in macrophages within coronary atherosclerotic plaques and epicardial/perivascular fat in autopsy cases and macrophages within thrombi covering ruptured coronary plaques in thrombectomy samples from patients with acute myocardial infarction. Second, we confirmed that FABP4 was secreted from macrophages and adipocytes cultured in vitro. Third, we investigated the effect of exogenous FABP4 on macrophages and human coronary artery-derived smooth muscle cells and endothelial cells in vitro. Treatment of the cells with recombinant FABP4 significantly increased gene expression of inflammatory markers in a dose-dependent manner. Finally, we measured serum FABP4 level in the aortic root (Ao-FABP4) and coronary sinus (CS-FABP4) of 34 patients with suspected or known coronary artery disease. Coronary stenosis score assessed by the modified Gensini score was weakly correlated with CS-FABP4 but was not correlated with Ao-FABP4. A stronger correlation (r=0.59, P<0.01) was observed for the relationship between coronary stenosis score and coronary veno-arterial difference in FABP4 level, (CS-Ao)-FABP4, indicating local production of FABP4 during coronary circulation in the heart. Multivariate analysis indicated that (CS-Ao)-FABP4 was an independent predictor of the severity of coronary stenosis after adjustment of conventional risk factors. Conclusions-FABP4 locally produced by epicardial/perivascular fat and macrophages in vascular plaques contributes to the development of coronary atherosclerosis.

植込み型除細動器導入症例におけるCKDの臨床的特徴

茂庭仁人, 瀬野結, 田中希尚, 古橋眞人, 吉田英昭, 藤戸健史, 望月敦史, 永原大五, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 58 ( 3 ) 280 - 280 2016.05

当院における植込み型除細動器導入透析患者の特徴

茂庭仁人, 瀬野結, 田中希尚, 古橋眞人, 吉田英昭, 藤戸健史, 望月敦史, 永原大五, 三浦哲嗣

日本透析医学会雑誌 ( (一社)日本透析医学会 ) 49 ( Suppl.1 ) 799 - 799 2016.05

Possible Increase in Serum FABP4 Level Despite Adiposity Reduction by Canagliflozin, an SGLT2 Inhibitor

Masato Furuhashi, Megumi Matsumoto, Shinya Hiramitsu, Akina Omori, Marenao Tanaka, Norihito Moniwa, Hideaki Yoshida, Junnichi Ishii, Tetsuji Miura

PLOS ONE ( PUBLIC LIBRARY SCIENCE ) 11 ( 4 ) 2016.04 [Refereed]

Background Fatty acid-binding protein 4 (FABP4/A-FABP/aP2) is secreted from adipocytes in association with catecholamine-induced lipolysis, and elevated serum FABP4 level is associated with obesity, insulin resistance and atherosclerosis. Secreted FABP4 as a novel adipokine leads to insulin resistance via increased hepatic glucose production (HGP). Sodium-glucose cotransporter 2 (SGLT2) inhibitors decrease blood glucose level via increased urinary glucose excretion, though HGP is enhanced. Here we investigated whether canagliflozin, an SGLT2 inhibitor, modulates serum FABP4 level. Methods Canagliflozin (100 mg/day) was administered to type 2 diabetic patients (n = 39) for 12 weeks. Serum FABP4 level was measured before and after treatment. Results At baseline, serum FABP4 level was correlated with adiposity, renal dysfunction and noradrenaline level. Treatment with canagliflozin significantly decreased adiposity and levels of fasting glucose and HbA1c but increased average serum FABP4 level by 10.3%(18.0 +/- 1.0 vs. 19.8 +/- 1.2 ng/ml, P = 0.008), though elevation of FABP4 level after treatment was observed in 26 (66.7%) out of 39 patients. Change in FABP4 level was positively correlated with change in levels of fasting glucose (r = 0.329, P = 0.044), HbA1c (r = 0.329, P = 0.044) and noradrenaline (r = 0.329, P = 0.041) but was not significantly correlated with change in adiposity or other variables. Conclusions Canagliflozin paradoxically increases serum FABP4 level in some diabetic patients despite amelioration of glucose metabolism and adiposity reduction, possibly via induction of catecholamine-induced lipolysis in adipocytes. Increased FABP4 level by canagliflozin may undermine the improvement of glucose metabolism and might be a possible mechanism of increased HGP by inhibition of SGLT2.

SGLT2阻害薬であるカナグリフロジンによる新規アディポカイン脂肪酸結合タンパク4(FABP4)の血中濃度への影響

古橋眞人, 松本めぐみ, 平光伸也, 大森晶奈, 田中希尚, 茂庭仁人, 吉田英昭, 石井潤一, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 59 ( Suppl.1 ) S - 336 2016.04

睡眠呼吸障害が血糖変動に与える影響の検討

中田圭, 丹野雅也, 矢野俊之, 小笠原淳, 赤坂憲, 古橋眞人, 三木隆幸, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 59 ( Suppl.1 ) S - 191 2016.04

Reduction of circulating FABP4 level by treatment with omega-3 fatty acid ethyl esters

Masato Furuhashi, Shinya Hiramitsu, Tomohiro Mita, Akina Omori, Takahiro Fuseya, Shutaro Ishimura, Yuki Watanabe, Kyoko Hoshina, Megumi Matsumoto, Marenao Tanaka, Norihito Moniwa, Hideaki Yoshida, Junnichi Ishii, Tetsuji Miura

LIPIDS IN HEALTH AND DISEASE ( BIOMED CENTRAL LTD ) 15 2016.01 [Refereed]

Background: Fatty acid-binding protein 4 (FABP4/A-FABP/aP2) mainly expressed in adipocytes is secreted and acts as an adipokine. Increased circulating FABP4 level is associated with obesity, insulin resistance and atherosclerosis. However, little is known about the modulation of serum FABP4 level by drugs including anti-dyslipidemic agents. Methods: Patients with dyslipidemia were treated with omega-3 fatty acid ethyl esters (4 g/day; n = 14) containing eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) for 4 weeks. Serum FABP4 level was measured before and after treatment. Expression and secretion of FABP4 were also examined in mouse 3T3-L1 adipocytes treated with EPA or DHA. Results: Treatment with omega-3 fatty acid ethyl esters significantly decreased triglycerides and serum FABP4 level (13.5 +/- 1.5 vs. 11.5 +/- 1.1 ng/ml, P = 0.017). Change in FABP4 level by omega-3 fatty acids was negatively correlated with change in levels of EPA + DHA (r = -0.643, P = 0.013), EPA (r = -0.540, P = 0.046) and DHA (r = -0.650, P = 0.011) but not change in the level of triglycerides or other fatty acid composition. Treatment of 3T3-L1 adipocytes with EPA or DHA had no effect on short-term (2 h) secretion of FABP4. However, gene expression and long-term (24 h) secretion of FABP4 were significantly reduced by treatment with EPA or DHA. Conclusions: Omega-3 fatty acids decrease circulating FABP4 level, possibly by reducing expression and consecutive secretion of FABP4 in adipocytes. Reducing FABP4 level might be involved in suppression of cardiovascular events by omega-3 fatty acids.

Reduction of serum FABP4 level by sitagliptin, a DPP-4 inhibitor, in patients with type 2 diabetes mellitus

Masato Furuhashi, Shinya Hiramitsu, Tomohiro Mita, Takahiro Fuseya, Shutaro Ishimura, Akina Omori, Megumi Matsumoto, Yuki Watanabe, Kyoko Hoshina, Marenao Tanaka, Norihito Moniwa, Hideaki Yoshida, Junnichi Ishii, Tetsuji Miura

JOURNAL OF LIPID RESEARCH ( AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC ) 56 ( 12 ) 2372 - 2380 2015.12 [Refereed]

Fatty acid binding protein 4 (FABP4), also known as adipocyte FABP or aP2, is secreted from adipocytes in association with lipolysis as a novel adipokine, and elevated serum FABP4 level is associated with obesity, insulin resistance, and atherosclerosis. However, little is known about the modulation of serum FABP4 level by therapeutic drugs. Sitagliptin (50 mg/day), a dipeptidyl peptidase 4 (DPP-4) inhibitor that increases glucagon-like peptide 1 (GLP-1), was administered to patients with type 2 diabetes (n = 24) for 12 weeks. Treatment with sitagliptin decreased serum FABP4 concentration by 19.7% (17.8 +/- 1.8 vs. 14.3 +/- 1.5 ng/ml, P < 0.001) and hemoglobin A1c without significant changes in adiposity or lipid variables. In 3T3-L1 adipocytes, sitagliptin or exendin-4, a GLP-1 receptor agonist, had no effect on short-term (2 h) secretion of FABP4. However, gene expression and long-term (24 h) secretion of FABP4 were significantly reduced by sitagliptin, which was not mimicked by exendin-4. Treatment with recombinant DPP-4 increased gene expression and long-term secretion of FABP4, and the effects were cancelled by sitagliptin. Furthermore, knockdown of DPP-4 in 3T3-L1 adipocytes decreased gene expression and long-term secretion of FABP4. In conclusion, sitagliptin decreases serum FABP4 level, at least in part, via reduction in the expression and consecutive secretion of FABP4 in adipocytes by direct inhibition of DPP-4.

シタグリプチンは新規アディポカイン脂肪酸結合タンパク4(FABP4)の血中濃度を低下させる

古橋眞人, 平光伸也, 石井潤一, 美田知宏, 大森晶奈, 伏屋敬博, 石村周太郎, 渡邊礼, 松本めぐみ, 田中希尚, 茂庭仁人, 吉田英昭, 三浦哲嗣

肥満研究 ( (一社)日本肥満学会 ) 21 ( Suppl. ) 175 - 175 2015.09

Angiotensin II receptor blockers decrease serum concentration of fatty acid-binding protein 4 in patients with hypertension

Masato Furuhashi, Tomohiro Mita, Norihito Moniwa, Kyoko Hoshina, Shutaro Ishimura, Takahiro Fuseya, Yuki Watanabe, Hideaki Yoshida, Kazuaki Shimamoto, Tetsuji Miura

HYPERTENSION RESEARCH ( NATURE PUBLISHING GROUP ) 38 ( 4 ) 252 - 259 2015.04 [Refereed]

Elevated circulating fatty acid-binding protein 4 (FABP4/A-FABP/aP2), an adipokine, is associated with obesity, insulin resistance, hypertension and cardiovascular events. However, how circulating FABP4 level is modified by pharmacological agents remains unclear. We here examined the effects of angiotensin II receptor blockers (ARBs) on serum FABP4 level. First, essential hypertensives were treated with ARBs: candesartan (8 mg day-1; n= 7) for 2 weeks, olmesartan (20 mg day-1; n= 9) for 12 weeks, and valsartan (80 mg day-1; n= 94) or telmisartan (40 mg day-1; n= 91) for 8 weeks added to amlodipine (5 mg day-1). Treatment with ARBs significantly decreased blood pressure and serum FABP4 concentrations by 8-20% without significant changes in adiposity or lipid variables, though the M value determined by hyperinsulinemic-euglycemic glucose clamp, a sensitive index of insulin sensitivity, was significantly increased by candesartan. Next, alterations in FABP4 secretion from 3T3-L1 adipocytes were examined under several agents. Lipolytic stimulation of the beta-adrenoceptor in 3T3-L1 adipocytes by isoproterenol increased FABP4 secretion, and conversely, insulin suppressed FABP4 secretion. However, treatment of 3T3-L1 adipocytes with angiotensin II or ARBs for 2 h had no effect on gene expression or secretion of FABP4 regardless of beta-adrenoceptor stimulation. In conclusion, treatment with structurally different ARBs similarly decreases circulating FABP4 concentrations in hypertensive patients as a class effect of ARBs, which is not attributable to blockade of the angiotensin II receptor in adipocytes. Reduction of FABP4 levels by ARBs might be involved in suppression of cardiovascular events.

オメガ-3脂肪酸エチルは新規アディポカイン脂肪酸結合タンパク4の血中濃度を低下させる

古橋眞人, 平光伸也, 石井潤一, 美田知宏, 大森晶奈, 伏屋敬博, 石村周太郎, 渡邊礼, 田中希尚, 吉田英昭, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 58 ( Suppl.1 ) S - 146 2015.04

FABP4 is Secreted from Adipocytes by Adenyl Cyclase-PKA- and Guanylyl Cyclase-PKG-Dependent Lipolytic Mechanisms

Tomohiro Mita, Masato Furuhashi, Shinya Hiramitsu, Junnichi Ishii, Kyoko Hoshina, Shutaro Ishimura, Takahiro Fuseya, Yuki Watanabe, Marenao Tanaka, Kohei Ohno, Hiroshi Akasaka, Hirofumi Ohnishi, Hideaki Yoshida, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

OBESITY ( WILEY-BLACKWELL ) 23 ( 2 ) 359 - 367 2015.02 [Refereed]

ObjectiveFatty acid-binding protein 4 (FABP4) is expressed in adipocytes, and elevated plasma FABP4 level is associated with obesity-mediated metabolic phenotype. Postprandial regulation and secretory signaling of FABP4 has been investigated. MethodsTime courses of FABP4 levels were examined during an oral glucose tolerance test (OGTT; n=53) or a high-fat test meal eating (n=35). Effects of activators and inhibitors of adenyl cyclase (AC)-protein kinase A (PKA) signaling and guanylyl cyclase (GC)-protein kinase G (PKG) signaling on FABP4 secretion from mouse 3T3-L1 adipocytes were investigated. ResultsFABP4 level significantly declined after the OGTT or a high-fat meal eating, while insulin level was increased. Treatment with low and high glucose concentration or palmitate for 2 h did not affect FABP4 secretion from 3T3-L1 adipocytes. FABP4 secretion was increased by stimulation of lipolysis using isoproterenol, a (3)-adrenoceptor agonist (CL316243), forskolin, dibutyryl-cAMP and atrial natriuretic peptide, and the induced FABP4 secretion was suppressed by insulin or an inhibitor of PKA (H-89), PKG (KT5823) or hormone sensitive lipase (CAY10499). ConclusionsFABP4 is secreted from adipocytes in association with lipolysis regulated by AC-PKA- and GC-PKG-mediated signal pathways. Plasma FABP4 level declines postprandially, and suppression of FABP4 secretion by insulin-induced anti-lipolytic signaling may be involved in this decline in FABP4 level.

Urinary Angiotensin-Converting Enzyme 2 in Hypertensive Patients May Be Increased by Olmesartan, an Angiotensin II Receptor Blocker

Masato Furuhashi, Norihito Moniwa, Tomohiro Mita, Takahiro Fuseya, Shutaro Ishimura, Kohei Ohno, Satoru Shibata, Marenao Tanaka, Yuki Watanabe, Hiroshi Akasaka, Hirofumi Ohnishi, Hideaki Yoshida, Hideki Takizawa, Shigeyuki Saitoh, Nobuyuki Ura, Kazuaki Shimamoto, Tetsuji Miura

AMERICAN JOURNAL OF HYPERTENSION ( OXFORD UNIV PRESS ) 28 ( 1 ) 15 - 21 2015.01 [Refereed]

BACKGROUND Angiotensin-converting enzyme 2 (ACE2) is highly expressed in the kidney and converts angiotensin (Ang) II to Ang-(1-7), a renoprotective peptide. Urinary ACE2 has been shown to be elevated in patients with chronic kidney disease. However, the effects of antihypertensive agents on urinary ACE2 remain unclear. METHODS Of participants in the Tanno-Sobetsu cohort study in 2011 (n = 617), subjects on no medication (n = 101) and hypertensive patients treated with antihypertensive agents, including the calcium channel blockers amlodipine and long-acting nifedipine; the ACE inhibitor enalapril; and the Ang II receptor blockers losartan, candesartan, valsartan, telmisartan, and olmesartan, for more than 1 year (n = 100) were enrolled, and urinary ACE2 level was measured. RESULTS Glucose and hemoglobin A1c were significantly higher in patients treated with enalapril, telmisartan or olmesartan than in the control subjects. Urinary albumin-to-creatinine ratio (UACR) was significantly higher in patients treated with enalapril than in the control subjects. Urinary ACE2 level was higher in the olmesartan-treated group, but not the other treatment groups, than in the control group. Urinary ACE2 level was positively correlated with systolic blood pressure (r = 0.211; P = 0.003), UACR (r = 0.367; P < 0.001), and estimated salt intake (r = 0.260; P < 0.001). Multivariable regression analysis after adjustment of age, sex, and the correlated indices showed that the use of olmesartan was an independent predictor of urinary ACE2 level. CONCLUSIONS In contrast with other antihypertensive drugs, olmesartan may uniquely increase urinary ACE2 level, which could potentially offer additional renoprotective effects.

Fatty acid-binding protein 4 (FABP4): Pathophysiological insights and potent clinical biomarker of metabolic and cardiovascular diseases

Masato Furuhashi, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

Clinical Medicine Insights: Cardiology ( Libertas Academica Ltd. ) 2014 23 - 33 2014.12 [Refereed]

Over the past decade, evidences of an integration of metabolic and inflammatory pathways, referred to as metaflammation in several aspects of metabolic syndrome, have been accumulating. Fatty acid-binding protein 4 (FABP4), also known as adipocyte FABP (A-FABP) or aP2, is mainly expressed in adipocytes and macrophages and plays an important role in the development of insulin resistance and atherosclerosis in relation to metaflam-mation. Despite lack of a typical secretory signal peptide, FABP4 has been shown to be released from adipocytes in a non-classical pathway associated with lipolysis, possibly acting as an adipokine. Elevation of circulating FABP4 levels is associated with obesity, insulin resistance, diabetes mellitus, hyperten-sion, cardiac dysfunction, atherosclerosis, and cardiovascular events. Furthermore, ectopic expression and function of FABP4 in several types of cells and tissues have been recently demonstrated. Here, we discuss both the significant role of FABP4 in pathophysiological insights and its usefulness as a biomarker of metabolic and cardiovascular diseases.

Urinary Excretion of Fatty Acid-Binding Protein 4 is Associated with Albuminuria and Renal Dysfunction

Yusuke Okazaki, Masato Furuhashi, Marenao Tanaka, Tomohiro Mita, Takahiro Fuseya, Shutaro Ishimura, Yuki Watanabe, Kyoko Hoshina, Hiroshi Akasaka, Hirofumi Ohnishi, Hideaki Yoshida, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

PLOS ONE ( PUBLIC LIBRARY SCIENCE ) 9 ( 12 ) e115429 2014.12 [Refereed]

Background: Fatty acid-binding protein 4 (FABP4/A-FABP/aP2) is expressed in not only adipocytes and macrophages but also peritubular capillaries in the normal kidney. We recently demonstrated that ectopic expression of FABP4, but not FABP1 known as liver FABP (L-FABP), in the glomerulus is associated with progression of proteinuria and renal dysfunction. However, urinary excretion of FABP4 has not been investigated. Methods: Subjects who participated in the Tanno-Sobetsu Study, a study with a population-based cohort design, in 2011 (n=392, male/female: 166/226) were enrolled. Urinary FABP4 (U-FABP4) and urinary albumin-to-creatinine ratio (UACR) were measured. Change in estimated glomerular filtration rate (eGFR) was followed up one year later. Results: In 93 (23.7%) of the 392 subjects, U-FABP4 level was below the sensitivity of the assay. Subjects with undetectable U-FABP4 were younger and had lower UACR and higher eGFR levels than subjects with measurable U-FABP4. U-FABP4 level was positively correlated with age, systolic blood pressure and levels of serum FABP4 (S-FABP4), triglycerides, hemoglobin A1c (HbA1c), urinary FABP1 (U-FABP1) and UACR (r=0.360, p<0.001). Age, S-FABP4, U-FABP1 and UACR were independent predictors of U-FABP4. On the other hand, systolic blood pressure, HbA1c and U-FABP4 were independently correlated with UACR. Reduction in eGFR after one year was significantly larger in a group with the highest tertile of baseline U-FABP4 than a group with the lowest tertile. Conclusions: Urinary FABP4 level is independently correlated with level of albuminuria and possibly predicts yearly decline of eGFR. U-FABP4 would be a novel biomarker of glomerular damage.

Reduction of endoplasmic reticulum stress inhibits neointima formation after vascular injury

Shutaro Ishimura, Masato Furuhashi, Tomohiro Mita, Takahiro Fuseya, Yuki Watanabe, Kyoko Hoshina, Nobuaki Kokubu, Katsumi Inoue, Hideaki Yoshida, Tetsuji Miura

SCIENTIFIC REPORTS ( NATURE PUBLISHING GROUP ) 4 6943 2014.11 [Refereed]

Endoplasmic reticulum (ER) stress and inappropriate adaptation through the unfolded protein response (UPR) are predominant features of pathological processes. However, little is known about the link between ER stress and endovascular injury. We investigated the involvement of ER stress in neointima hyperplasia after vascular injury. The femoral arteries of 7-8-week-old male mice were subjected to wire-induced vascular injury. After 4 weeks, immunohistological analysis showed that ER stress markers were upregulated in the hyperplastic neointima. Neointima formation was increased by 54.8% in X-box binding protein-1 (XBP1) heterozygous mice, a model of compromised UPR. Knockdown of Xbp1 in human coronary artery smooth muscle cells (CASMC) in vitro promoted cell proliferation and migration. Furthermore, treatment with ER stress reducers, 4-phenylbutyrate (4-PBA) and tauroursodeoxycholic acid (TUDCA), decreased the intima-to-media ratio after wire injury by 50.0% and 72.8%, respectively. Chronic stimulation of CASMC with PDGF-BB activated the UPR, and treatment with 4-PBA and TUDCA significantly suppressed the PDGF-BB-induced ER stress markers in CASMC and the proliferation and migration of CASMC. In conclusion, increased ER stress contributes to neointima formation after vascular injury, while UPR signaling downstream of XBP1 plays a suppressive role. Suppression of ER stress would be a novel strategy against post-angioplasty vascular restenosis.

脂肪酸結合タンパク4はAC-PKAおよびGC-PKG経路を介して脂肪分解と共に脂肪細胞から分泌される

美田知宏, 古橋眞人, 石村周太郎, 伏屋敬博, 保科京香, 渡邊礼, 大野紘平, 田中希尚, 吉田英昭, 三浦哲嗣

肥満研究 ( (一社)日本肥満学会 ) 20 ( Suppl. ) 161 - 161 2014.10

アンジオテンシンII受容体拮抗薬による新規アディポカイン脂肪酸結合タンパク4濃度の変化

伏屋敬博, 古橋眞人, 美田知宏, 茂庭仁人, 渡邊礼, 大森晶奈, 大野紘平, 田中希尚, 石村周太郎, 吉田英昭, 三浦哲嗣

日本高血圧学会総会プログラム・抄録集 ( (NPO)日本高血圧学会 ) 37回 391 - 391 2014.10

Emerging issues in radiogenic cataracts and cardiovascular disease

Nobuyuki Hamada, Yuki Fujimichi, Toshiyasu Iwasaki, Noriko Fujii, Masato Furuhashi, Eri Kubo, Tohru Minamino, Takaharu Nomura, Hitoshi Sato

JOURNAL OF RADIATION RESEARCH ( OXFORD UNIV PRESS ) 55 ( 5 ) 831 - 846 2014.09 [Refereed]

In 2011, the International Commission on Radiological Protection issued a statement on tissue reactions (formerly termed non-stochastic or deterministic effects) to recommend lowering the threshold for cataracts and the occupational equivalent dose limit for the crystalline lens of the eye. Furthermore, this statement was the first to list circulatory disease (cardiovascular and cerebrovascular disease) as a health hazard of radiation exposure and to assign its threshold for the heart and brain. These changes have stimulated various discussions and may have impacts on some radiation workers, such as those in the medical sector. This paper considers emerging issues associated with cataracts and cardiovascular disease. For cataracts, topics dealt with herein include (i) the progressive nature, stochastic nature, target cells and trigger events of lens opacification, (ii) roles of lens protein denaturation, oxidative stress, calcium ions, tumor suppressors and DNA repair factors in cataractogenesis, (iii) dose rate effect, radiation weighting factor, and classification systems for cataracts, and (iv) estimation of the lens dose in clinical settings. Topics for cardiovascular disease include experimental animal models, relevant surrogate markers, latency period, target tissues, and roles of inflammation and cellular senescence. Future research needs are also discussed.

Elevation of circulating fatty acid-binding protein 4 is independently associated with left ventricular diastolic dysfunction in a general population

Takahiro Fuseya, Masato Furuhashi, Satoshi Yuda, Atsuko Muranaka, Mina Kawamukai, Tomohiro Mita, Shutaro Ishimura, Yuki Watanabe, Kyoko Hoshina, Marenao Tanaka, Kohei Ohno, Hiroshi Akasaka, Hirofumi Ohnishi, Hideaki Yoshida, Shigeyuki Saitoh, Kazuaki Shimamoto, Tetsuji Miura

CARDIOVASCULAR DIABETOLOGY ( BIOMED CENTRAL LTD ) 13 126 2014.08 [Refereed]

Background: Fatty acid-binding protein 4 (FABP4) is expressed in both adipocytes and macrophages. Recent studies have shown secretion of FABP4 from adipocytes and association of elevated serum FABP4 level with obesity, insulin resistance, hypertension, and atherosclerosis. However, little is known about role of FABP4 in cardiac function. Methods: From the database of the Tanno-Sobetsu Study, data for 190 subjects (male/female: 82/108) who were not treated with any medication and underwent echocardiography in 2011 or 2012 were retrieved for analyses of relationships between serum FABP4 concentration, metabolic markers and parameters of echocardiography. Results: Serum FABP4 level was positively correlated with age, body mass index (BMI), blood pressure (BP), LDL cholesterol, HOMA-R and mean left ventricular (LV) wall thickness (LVWT, males: r = 0.315, females: r = 0.401, p < 0.01) and was negatively correlated with HDL cholesterol, estimated glomerular filtration rate (eGFR) and peak myocardial velocity during early diastole (e'; males: r = -0.434, females: r = -0.353, p < 0.01), an index of LV diastolic function. However, no significant correlation was found between FABP4 level and LV end-diastolic dimension, LV ejection fraction or LV mass index. There were significant correlations of e' with age, BMI, BP, eGFR, brain natriuretic peptide (BNP), FABP4, metabolic markers and LVWT. Multivariate regression analysis adjusted by HOMA-R, BMI, eGFR, BNP or LVWT in addition to age, gender and BP revealed that serum FABP4 concentration was independently correlated with e'. Conclusions: Elevation of circulating FABP4 may contribute to LV diastolic dysfunction in a general population.

IgA型多発性骨髄腫に伴うacute light chain tubulopathyの一例

瀬野結, 田中希尚, 舘越勇輝, 青山ちひろ, 大野紘平, 古橋眞人, 吉田英昭, 坂本孝志, 小川弥生

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 56 ( 6 ) 877 - 877 2014.08

Reduction of endoplasmic reticulum stress by 4-phenylbutyric acid prevents the development of hypoxia-induced pulmonary arterial hypertension

Masayuki Koyama, Masato Furuhashi, Shutaro Ishimura, Tomohiro Mita, Takahiro Fuseya, Yusuke Okazaki, Hideaki Yoshida, Kazufumi Tsuchihashi, Tetsuji Miura

AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY ( AMER PHYSIOLOGICAL SOC ) 306 ( 9 ) H1314 - H1323 2014.05 [Refereed]

Pulmonary arterial hypertension (PAH) is characterized by vasoconstriction and vascular remodeling of the pulmonary artery (PA). Recently, endoplasmic reticulum (ER) stress and inappropriate adaptation through the unfolded protein response (UPR) have been disclosed in various types of diseases. Here we examined whether ER stress is involved in the pathogenesis of PAH. Four weeks of chronic normobaric hypoxia increased right ventricular (RV) systolic pressure by 63% compared with that in normoxic controls and induced RV hypertrophy and medial thickening of the PA in C57BL/6J mice. Treatment with 4-phenylbutyric acid (4-PBA), a chemical chaperone, significantly reduced RV systolic pressure by 30%, attenuated RV hypertrophy and PA muscularization, and increased total running distance in a treadmill test by 70% in hypoxic mice. The beneficial effects of 4-PBA were associated with suppressed expression of inflammatory cytokines and ER stress markers, including Grp78 and Grp94 in the activating transcription factor-6 branch, sXbp1 and Pdi in the inositol-requiring enzyme-1 branch and Atf4 in the PKR-like ER kinase branch, and reduced phosphorylation of c-Jun NH2-terminal kinase and eukaryotic translation initiation factor-2 alpha in the lung. The pattern of changes in ER stress and inflammatory markers by 4-PBA in the lung of the PAH model was reproduced in PA smooth muscle cells by chronic stimulation of platelet-derived growth factor-BB or hypoxia. Furthermore, knockdown of each UPR branch sensor activated other branches and promoted proliferation of PA smooth muscle cells. The findings indicate that activation of all branches of the UPR and accompanying inflammation play a major role in the pathogenesis of PAH, and that chemical chaperones are potentially therapeutic agents for PAH.

Small-Molecule Inhibitors of PKR Improve Glucose Homeostasis in Obese Diabetic Mice

Takahisa Nakamura, Alessandro Arduini, Brenna Baccaro, Masato Furuhashi, Goekhan S. Hotamisligil

DIABETES ( AMER DIABETES ASSOC ) 63 ( 2 ) 526 - 534 2014.02 [Refereed]

Obesity and metabolic diseases appear as clusters, often featuring high risk for insulin resistance and type 2 diabetes, and constitute a major global health problem with limited treatment options. Previous studies have shown that double-stranded RNA-dependent kinase, PKR, plays an important role in the nutrient/pathogen-sensing interface, and acts as a key modulator of chronic metabolic inflammation, insulin sensitivity, and glucose homeostasis in obesity. Recently, pathological PKR activation was also demonstrated in obese humans, strengthening its prospects as a potential drug target. Here, we investigate the use of two structurally distinct small-molecule inhibitors of PKR in the treatment of insulin resistance and type 2 diabetes in cells and in a mouse model of severe obesity and insulin resistance. Inhibition of PKR reduced stress-induced Jun NH2-terminal kinase activation and insulin receptor substrate 1 serine phosphorylation in vitro and in vivo. In addition, treatment with both PKR inhibitors reduced adipose tissue inflammation, improved insulin sensitivity, and improved glucose intolerance in mice after the establishment of obesity and insulin resistance. Our findings suggest that pharmacologically targeting PKR may be an effective therapeutic strategy for the treatment of insulin resistance and type 2 diabetes.

Ectopic expression of fatty acid-binding protein 4 in the glomerulus is associated with proteinuria and renal dysfunction.

Marenao Tanaka, Masato Furuhashi, Yusuke Okazaki, Tomohiro Mita, Takahiro Fuseya, Kohei Ohno, Shutaro Ishimura, Hideaki Yoshida, Tetsuji Miura

Nephron. Clinical practice ( 3-4 ) 128 ( 3-4 ) 345 - 51 2014 [Refereed] [International journal]

BACKGROUND/AIMS: Fatty acid-binding proteins (FABPs) are a family of intracellular lipid chaperones. Among FABPs, FABP1 (liver FABP) is expressed in proximal tubular epithelial cells in the kidney, and urinary FABP1 has been reported to reflect damage of proximal tubular epithelial cells. However, roles of other FABP isoforms in renal pathologies have not been reported. Recently, FABP4 (adipocyte FABP/aP2) was reported to be expressed in peritubular capillaries (PTCs), but not in glomerular capillaries in the normal kidney. We examined the hypothesis that pathological conditions alter the level and localization of FABP4 expression in the kidney, which mediates renal dysfunction. METHODS: A total of 112 consecutive patients who underwent renal biopsy were retrospectively enrolled. Expression of FABP4 protein and mRNA in the kidney was examined by immunohistochemistry and in situ hybridization, respectively. The ratio of FABP4-positive area to total area within glomeruli (G-FABP4-Area), urinary protein level (U-Protein), and change in estimated glomerular filtration rate (eGFR) 1 year after biopsy were examined. RESULTS: FABP4 protein and mRNA were expressed not only in PTCs, but also in endothelial cells and macrophages in the glomerulus. G-FABP4-Area was correlated with U-Protein (r = 0.497, p < 0.001). As a subanalysis, in patients with IgA nephropathy (n = 34), G-FABP4-Area was significantly larger in cases with an endocapillary proliferative lesion, and change in eGFR was negatively correlated with G-FABP4-Area at baseline (r = -0.537, p = 0.008). CONCLUSION: Ectopic FABP4 expression in the glomerulus is induced by renal diseases and is closely associated with proteinuria and renal dysfunction.

Ectopic Expression of Fatty Acid-Binding Protein 4 in the Glomerulus Is Associated with Proteinuria and Renal Dysfunction

Marenao Tanaka, Masato Furuhashi, Yusuke Okazaki, Tomohiro Mita, Takahiro Fuseya, Kohei Ohno, Shutaro Ishimura, Hideaki Yoshida, Tetsuji Miura

NEPHRON CLINICAL PRACTICE ( KARGER ) 128 ( 3-4 ) 345 - 351 2014 [Refereed]

Background/Aims: Fatty acid-binding proteins (FABPs) are a family of intracellular lipid chaperones. Among FABPs, FABP1 (liver FABP) is expressed in proximal tubular epithelial cells in the kidney, and urinary FABP1 has been reported to reflect damage of proximal tubular epithelial cells. However, roles of other FABP isoforms in renal pathologies have not been reported. Recently, FABP4 (adipocyte FABP/aP2) was reported to be expressed in peritubular capillaries (PTCs), but not in glomerular capillaries in the normal kidney. We examined the hypothesis that pathological conditions alter the level and localization of FABP4 expression in the kidney, which mediates renal dysfunction. Methods: A total of 112 consecutive patients who underwent renal biopsy were retrospectively enrolled. Expression of FABP4 protein and mRNA in the kidney was examined by immunohistochemistry and in situ hybridization, respectively. The ratio of FABP4-positive area to total area within glomeruli (G-FABP4-Area), urinary protein level (U-Protein), and change in estimated glomerular filtration rate (eGFR) 1 year after biopsy were examined. Results: FABP4 protein and mRNA were expressed not only in PTCs, but also in endothelial cells and macrophages in the glomerulus. G-FABP4-Area was correlated with U-Protein (r = 0.497, p < 0.001). As a subanalysis, in patients with IgA nephropathy (n = 34), G-FABP4-Area was significantly larger in cases with an endocapillary proliferative lesion, and change in eGFR was negatively correlated with G-FABP4-Area at baseline (r = -0.537, p = 0.008). Conclusion: Ectopic FABP4 expression in the glomerulus is induced by renal diseases and is closely associated with proteinuria and renal dysfunction. (C) 2015 S. Karger AG, Basel

Circulating Levels of Fatty Acid-Binding Protein Family and Metabolic Phenotype in the General Population

Shutaro Ishimura, Masato Furuhashi, Yuki Watanabe, Kyoko Hoshina, Takahiro Fuseya, Tomohiro Mita, Yusuke Okazaki, Masayuki Koyama, Marenao Tanaka, Hiroshi Akasaka, Hirofumi Ohnishi, Hideaki Yoshida, Shigeyuki Saitoh, Tetsuji Miura

PLOS ONE ( PUBLIC LIBRARY SCIENCE ) 8 ( 11 ) e81318 2013.11 [Refereed]

Objective: Fatty acid-binding proteins (FABPs) are a family of 14-15-kDa proteins, and some FABPs have been to be used as biomarkers of tissue injury by leak from cells. However, recent studies have shown that FABPs can be secreted from cells into circulation. Here we examined determinants and roles of circulating FABPs in a general population. Methods: From the database of the Tanno-Sobetsu Study, a study with a population-based cohort design, data in 2011 for 296 subjects on no medication were retrieved, and FABP1 similar to 5 in their serum samples were assayed. Results: Level of FABP4, but not the other isoforms, showed a gender difference, being higher in females than in males. Levels of all FABPs were negatively correlated with estimated glomerular filtration rate (eGFR), but a distinct pattern of correlation with other clinical parameters was observed for each FABP isoform; significant correlates were alanine aminotransferase (ALT), blood pressure (BP), and brain natriuretic peptide (BNP) for FABP1, none besides eGFR for FABP2, age, BP, and BNP for FABP3, age, waist circumference (WC), BP, BNP, lipid variables, high-sensitivity C-reactive protein (hsCRP), and HOMA-R for FABP4, and age, WC, BP, ALT, BNP, and HOMA-R for FABP5. FABP4 is the most strongly related to metabolic markers among FABPs. In a multivariate regression analysis, FABP4 level was an independent predictor of HOMAR after adjustment of age, gender, WC, BP, HDL cholesterol, and hsCRP. Conclusions: Each FABP isoform level showed a distinct pattern of correlation with clinical parameters, although levels of all FABPs were negatively determined by renal function. Circulating FABP4 appears to be a useful biomarker for detecting preclinical stage of metabolic syndrome, especially insulin resistance, in the general population.

脂肪酸結合タンパク4の脂肪細胞からの分泌機構の検討

石村周太郎, 古橋眞人, 美田知宏, 伏屋敬博, 大野紘平, 田中希尚, 赤坂憲, 吉田英昭, 平光伸也, 石井潤一, 三浦哲嗣

肥満研究 ( (一社)日本肥満学会 ) 19 ( Suppl. ) 176 - 176 2013.09

メサンギウム増殖性糸球体腎炎における糸球体脂肪酸結合タンパク4発現の意義

田中希尚, 古橋眞人, 岡崎雄介, 大野紘平, 美田知宏, 伏屋敬博, 石村周太郎, 吉田英昭, 三浦哲嗣

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 55 ( 6 ) 1065 - 1065 2013.08

Adipocyte lipid chaperone aP2 Is a secreted adipokine regulating hepatic glucose production

Haiming Cao, Motohiro Sekiya, Meric Erikci Ertunc, M. Furkan Burak, Jared R. Mayers, Ariel White, Karen Inouye, Lisa M. Rickey, Baris C. Ercal, Masato Furuhashi, Gürol Tuncman, Gökhan S. Hotamisligil

Cell Metabolism ( 5 ) 17 ( 5 ) 768 - 778 2013.05 [Refereed]

Proper control of hepatic glucose production is central to whole-body glucose homeostasis, and its disruption plays a major role in diabetes. Here, we demonstrate that although established as an intracellular lipid chaperone, aP2 is in fact actively secreted from adipocytes to control liver glucose metabolism. Secretion of aP2 from adipocytes is regulated by fasting- and lipolysis-related signals, and circulating aP2 levels are markedly elevated in mouse and human obesity. Recombinant aP2 stimulates glucose production and gluconeogenic activity in primary hepatocytes in vitro and in lean mice in vivo. In contrast, neutralization of secreted aP2 reduces glucose production and corrects the diabetic phenotype of obese mice. Hyperinsulinemic-euglycemic and pancreatic clamp studies upon aP2 administration or neutralization demonstrated actions of aP2 in liver. We conclude that aP2 is an adipokine linking adipocytes to hepatic glucose production and that neutralizing secreted aP2 may represent an effective therapeutic strategy against diabetes. © 2013 Elsevier Inc.

急激な血糖悪化を契機に偶然発見され、多腺性内分泌腫瘍症1型が疑われた1例

菅原浩仁, 小笠原惇, 伊藤孝仁, 田中希尚, 古橋眞人, 下重晋也, 丹野雅也, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 56 ( 1 ) 41 - 41 2013.01

Histopathology of the pancreas in fulminant type 1 diabetes after 23-year follow-up: a case report

Tatsuya Sato, Takayuki Miki, Naoto Murakami, Yoshihiko Hirohashi, Hidemichi Kouzu, Masato Furuhashi, Masaya Tanno, Satoshi Yuda, Shigeyuki Saitoh, Tadashi Hasegawa, Tetsuji Miura

PATHOLOGY INTERNATIONAL ( WILEY-BLACKWELL ) 62 ( 12 ) 827 - 829 2012.12 [Refereed]

Elevation of Fatty Acid-Binding Protein 4 Is Predisposed by Family History of Hypertension and Contributes to Blood Pressure Elevation

Hideki Ota, Masato Furuhashi, Shutaro Ishimura, Masayuki Koyama, Yusuke Okazaki, Tomohiro Mita, Takahiro Fuseya, Tomohisa Yamashita, Marenao Tanaka, Hideaki Yoshida, Kazuaki Shimamoto, Tetsuji Miura

AMERICAN JOURNAL OF HYPERTENSION ( OXFORD UNIV PRESS ) 25 ( 10 ) 1124 - 1130 2012.10 [Refereed]

BACKGROUND Fatty acid-binding protein 4 (FABP4/A-FABP/aP2), a lipid chaperone, is expressed in both adipocytes and macrophages. Recent studies have shown secretion of FABP4 from adipocytes and association of elevated serum FABP4 level with obesity, insulin resistance, and atherosclerosis. However, little is known about the role of FABP4 in essential hypertension. METHODS We first examined serum FABP4 concentrations in 18 normotensives (NT) and 30 nontreated essential hypertensives (EHT). The EHT were divided into 18 insulin-sensitive EHT (EHT-S) and 12 insulin-resistant EHT (EHT-R) based on their insulin-sensitivity index, the M value, determined by the hyperinsulinemic euglycemic clamp technique. In the second study, we determined FABP4 levels in 30 young NT men with or without a family history of hypertension (FH+ and FH-, respectively; n = 15 each). RESULTS Serum FABP4 level was significantly higher in the EHT-R than in the NT, whereas elevation of FABP4 level in the EHT-S was not statistically significant. FABP4 level was positively correlated with age, body mass index (BMI), blood pressure, and triglycerides and negatively correlated with the M value. FABP4 level was an independent predictor of mean arterial pressure after adjustment of age, gender, and adiposity. The FH+ group had a significantly lower level of M value and higher level of FABP4 than did the FH(-)group, and FABP4 concentration was an independent determinant of the M value. CONCLUSIONS FABP4 contributes to blood pressure elevation and atherogenic metabolic phenotype in hypertensives, and the elevation of FABP4 is predisposed by a family history of hypertension.

妊娠関連発症劇症1型糖尿病の発症後23年が経過し、日和見感染症で死亡した一剖検例

村上直人, 佐藤達也, 廣橋良彦, 神津英至, 古橋眞人, 湯田聡, 三木隆幸, 斎藤重幸, 土橋和文, 三浦哲嗣

糖尿病 ( (一社)日本糖尿病学会 ) 55 ( 10 ) 827 - 827 2012.10

Angiotensin II receptor activation in youth triggers persistent insulin resistance and hypertension-a legacy effect?

Nobuhiko Togashi, Takuto Maeda, Hideaki Yoshida, Masayuki Koyama, Marenao Tanaka, Masato Furuhashi, Kazuaki Shimamoto, Tetsuji Miura

HYPERTENSION RESEARCH ( NATURE PUBLISHING GROUP ) 35 ( 3 ) 334 - 340 2012.03 [Refereed]

Although the involvement of angiotensin II (Ang II) in insulin resistance and hypertension has been established, the temporal relationships between Ang II receptor activation and changes in insulin sensitivity and blood pressure are not clear. To better understand this issue, we infused rats with Ang II (200 ng kg(-1) min(-1)) or vehicle for 4 weeks and assessed the residual effects after the discontinuation of the infusion on blood pressure, insulin sensitivity and tissue parameters of inflammation. Four weeks after the discontinuation of the Ang II infusion, the blood pressure was higher by 12.8mmHg, and insulin sensitivity as determined by a euglycemic hyperinsulinemic glucose clamp was reduced (glucose infusion rate: 11.1+/-0.7 vs. 17.6+/-0.5mg kg(-1) min(-1)) in the Ang II-treated group compared with controls. The persistent hypertension and insulin resistance were associated with greater than two-fold increases in macrophage chemoattractant protein-1, tumor necrosis factor-alpha and thiobarbituric acid-reactive substrates in the soleus muscle. Furthermore, total and activated forms of Rac-1, a regulatory subunit of the NADPH oxidase complex, were increased by 144 +/- 14% and 277 +/- 82%, respectively, in the skeletal muscle of Ang II-treated rats. These residual effects after Ang II infusion were all attenuated by the co-administration of tempol, a free radical scavenger, or candesartan with Ang II. The effects of candesartan were not mimicked by hydralazine at an equidepressant dose. These findings suggest that Ang II receptor activation in youth triggers the upregulation of inflammatory cytokines and the production of reactive oxygen species, thereby inducing later insulin resistance and hypertension. Hypertension Research (2012) 35, 334-340; doi: 10.1038/hr.2011.206; published online 1 December 2011

Serum Fatty Acid-Binding Protein 4 Is a Predictor of Cardiovascular Events in End-Stage Renal Disease

Masato Furuhashi, Shutaro Ishimura, Hideki Ota, Manabu Hayashi, Takahiro Nishitani, Marenao Tanaka, Hideaki Yoshida, Kazuaki Shimamoto, Goekhan S. Hotamisligil, Tetsuji Miura

PLOS ONE ( PUBLIC LIBRARY SCIENCE ) 6 ( 11 ) e27356 2011.11 [Refereed]

Background: Fatty acid-binding protein 4 (FABP4/A-FABP/aP2), a lipid chaperone, is expressed in both adipocytes and macrophages. Recent studies have shown that FABP4 is secreted from adipocytes and that FABP4 level is associated with obesity, insulin resistance, and atherosclerosis. However, little is known about the impact of FABP4 concentrations on prognosis. We tested the hypothesis that FABP4 level predicts prognosis of patients with end-stage renal disease (ESRD), a group at high risk for atherosclerosis-associated morbidity and mortality. Methods and Results: Biochemical markers including FABP4 were determined in 61 ESRD patients on chronic hemodialysis (HD). Serum FABP4 level in females (404.2 +/- 30.5 ng/ml) was significantly higher than that in males (315.8 +/- 30.0 ng/ml), and the levels in ESRD patients were about 20-times higher than those in age-, gender- and body mass index (BMI)-matched control subjects with normal renal function. FABP4 level was decreased by 57.2% after HD and was positively correlated with blood pressure, BMI, and levels of lipids and insulin. Multiple regression analysis indicated that HD duration, BMI, and triglycerides level were independent determinants for FABP4 level. ESRD patients with high FABP4 levels had higher cardiovascular mortality during the 7-year follow-up period. Cox proportional hazard regression analysis showed that logarithmically transformed FABP4 level was an independent predictor of cardiovascular death adjusted for age, gender, HD duration, BMI, and triglycerides level (hazard ratio, 7.75; 95% CI, 1.05-25.31). Conclusion: These findings suggest that FABP4 level, being related to adiposity and metabolic disorders, is a novel predictor of cardiovascular mortality in ESRD.

Deterioration of Renal Function by Chronic Heart Failure is Associated with Congestion and Oxidative Stress in the Tubulointerstitium

Marenao Tanaka, Hideaki Yoshida, Masato Furuhashi, Nobuhiko Togashi, Masayuki Koyama, Satoshi Yamamoto, Tomohisa Yamashita, Yusuke Okazaki, Shutaro Ishimura, Hideki Ota, Tadashi Hasegawa, Tetsuji Miura

INTERNAL MEDICINE ( JAPAN SOC INTERNAL MEDICINE ) 50 ( 23 ) 2877 - 2887 2011 [Refereed]

Objective We examined the hypothesis that renal congestion is responsible for the decline in renal function in patients with heart failure (HF) via tubulointerstitial inflammation. Methods First, in a longitudinal study, we retrospectively examined the relationship between cardiac functions and the decline of renal function during a period of 6.4 +/- 3.2 years in 20 patients who had a left ventricular ejection fraction of <40% and an estimated glomerular filtration ratio (eGFR) of <60 mL/min/1.73 m(2). Second, we compared the renal histology in autopsy cases of HF-induced renal dysfunction (HF-RD), cases of nephrosclerosis (NSC) and cases of neither RD nor HF (controls) in a cross-sectional study. Third, we retrospectively examined renal function in HF patients with predominantly right, but not left, ventricular dysfunction. Results eGFR decreased at 9.4 +/- 4.6%/year in the cohort of the longitudinal study. The rate of eGFR decline was correlated with blood pressure and with diameter of the inferior vena cava (IVCd) (r=0.5) measured at the initial work-up. Multivariate analysis indicated that the IVCd is an independent determinant of decline of eGFR in HF. In the cross-sectional study, fibrosis, Rac1 expression, protein nitrosylation, and number of CD68-positive cells were increased in the tubulointerstitium in both cases of HF-RD and NSC. Peritubular capillaries in HF-RD were dilated by 35% without any change in density compared with those in the controls. In right sided HF, the reduction of IVCd after treatment was associated with improvement of eGFR. Conclusion Venous congestion may contribute to HF-induced deterioration of renal function by augmenting oxidative stress-mediated inflammation in the tubulointerstitium.

Lipid chaperones and metabolic inflammation.

Furuhashi M, Ishimura S, Ota H, Miura T

International journal of inflammation 2011 642612 2011 [Refereed]

腎に限局したIgG4関連疾患の一例

岡崎雄介, 田中希尚, 山本諭, 山下智久, 萩原誠, 富樫信彦, 古橋眞人, 吉田英昭, 三浦哲嗣, 山本元久, 苗代康可

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 52 ( 6 ) 704 - 704 2010.08

慢性心不全による腎障害進展の機序にはうっ血性障害が関与する

田中希尚, 吉田英昭, 富樫信彦, 山下智久, 岡崎雄介, 石村周太郎, 茂庭仁人, 古橋眞人, 島本和明

日本腎臓学会誌 ( (一社)日本腎臓学会 ) 52 ( 3 ) 285 - 285 2010.05

Double-Stranded RNA-Dependent Protein Kinase Links Pathogen Sensing with Stress and Metabolic Homeostasis

Takahisa Nakamura, Masato Furuhashi, Ping Li, Haiming Cao, Gurol Tuncman, Nahum Sonenberg, Cem Z. Gorgun, Goekhan S. Hotamisligil

CELL ( CELL PRESS ) 140 ( 3 ) 338 - U41 2010.02 [Refereed]

As chronic inflammation is a hallmark of obesity, pathways that integrate nutrient- and pathogen sensing pathways are of great interest in understanding the mechanisms of insulin resistance, type 2 diabetes, and other chronic metabolic pathologies. Here, we provide evidence that double-stranded RNA-dependent protein kinase (PKR) can respond to nutrient signals as well as endoplasmic reticulum (ER) stress and coordinate the activity of other critical inflammatory kinases such as the c-Jun N-terminal kinase (JNK) to regulate insulin action and metabolism. PKR also directly targets and modifies insulin receptor substrate and hence integrates nutrients and insulin action with a defined pathogen response system. Dietary and genetic obesity features marked activation of PKR in adipose and liver tissues and absence of PKR alleviates metabolic deterioration due to nutrient or energy excess in mice. These findings demonstrate PKR as a critical component of an inflammatory complex that responds to nutrients and organelle dysfunction.

A Predominant Role for Parenchymal c-Jun Amino Terminal Kinase (JNK) in the Regulation of Systemic Insulin Sensitivity

Sara N. Vallerie, Masato Furuhashi, Raquel Fucho, Goekhan S. Hotamisligil

PLOS ONE ( PUBLIC LIBRARY SCIENCE ) 3 ( 9 ) e3151 2008.09 [Refereed]

It has been established that c-Jun N-terminal kinase 1 (JNK1) is essential to the pathogenesis of insulin resistance and type 2 diabetes. Although JNK influences inflammatory signaling pathways, it remains unclear whether its activity in macrophages contributes to adipose tissue inflammation and ultimately to the regulation of systemic metabolism. To address whether the action of this critical inflammatory kinase in bone marrow-derived elements regulates inflammatory responses in obesity and is sufficient and necessary for the deterioration of insulin sensitivity, we performed bone marrow transplantation studies with wild type and JNK1-deficient mice. These studies illustrated that JNK1-deficiency in the bone marrow-derived elements (BMDE) was insufficient to impact macrophage infiltration or insulin sensitivity despite modest changes in the inflammatory profile of adipose tissue. Only when the parenchymal elements lacked JNK1 could we demonstrate a significant increase in systemic insulin sensitivity. These data indicate that while the JNK1 activity in BMDE is involved in metabolic regulation and adipose milieu, it is epistatic to JNK1 activity in the parenchymal tissue for regulation of metabolic homeostasis.

Adipocyte/macrophage fatty acid-binding proteins contribute to metabolic deterioration through actions in both macrophages and adipocytes in mice

Masato Furuhashi, Raquel Fucho, Cem Z. Gorgun, Guerol Tuncman, Haiming Cao, Goekhan S. Hotamisligil

JOURNAL OF CLINICAL INVESTIGATION ( AMER SOC CLINICAL INVESTIGATION INC ) 118 ( 7 ) 2640 - 2650 2008.07 [Refereed]

Adipose tissue inflammation is a characteristic of obesity. However, the mechanisms that regulate this inflammatory response and link adipose inflammation to systemic metabolic consequences are not fully understood. In this study, we have taken advantage of the highly restricted coexpression of adipocyte/macrophage fatty acid-binding proteins (FABPs) aP2 (FABP4) and mal1 (FABP5) to examine the contribution of these lipid chaperones in macrophages and adipocytes to local and systemic inflammation and metabolic homeostasis in mice. Deletion of FABPs in adipocytes resulted in reduced expression of inflammatory cytokines in macrophages, whereas the same deletion in macrophages led to enhanced insulin signaling and glucose uptake in adipocytes. Using radiation chimerism through bone marrow transplantation, we generated mice with FABP deficiency in bone marrow and stroma-derived elements in vivo and studied the impact of each cellular target on local and systemic insulin action and glucose metabolism in dietary obesity. The results of these experiments indicated that neither macrophages nor adipocytes individually could account for the total impact of FABPs on systemic metabolism and suggest that interactions between these 2 cell types, particularly in adipose tissue, are critical for the inflammatory basis of metabolic deterioration.

Fatty acid-binding proteins: role in metabolic diseases and potential as drug targets

Masato Furuhashi, Goekhan S. Hotamisligil

NATURE REVIEWS DRUG DISCOVERY ( NATURE PUBLISHING GROUP ) 7 ( 6 ) 489 - 503 2008.06 [Refereed]

Lipids are vital components of many biological processes and crucial in the pathogenesis of numerous common diseases, but the specific mechanisms coupling intracellular lipids to biological targets and signalling pathways are not well understood. This is particularly the case for cells burdened with high lipid storage, trafficking and signalling capacity such as adipocytes and macrophages. Here, we discuss the central role of lipid chaperones-the fatty acid-binding proteins (FABPs)-in lipid-mediated biological processes and systemic metabolic homeostasis through the regulation of diverse lipid signals, and highlight their therapeutic significance. Pharmacological agents that modify FABP function may provide tissue-specific or cell-type-specific control of lipid signalling pathways, inflammatory responses and metabolic regulation, potentially providing a new class of drugs for diseases such as obesity, diabetes and atherosclerosis.

Treatment of diabetes and atherosclerosis by inhibiting fatty-acid-binding protein aP2

Masato Furuhashi, Guerol Tuncman, Cem Z. Goerguen, Liza Makowski, Genichi Atsumi, Eric Vaillancourt, Keita Kono, Vladimir R. Babaev, Sergio Fazio, MacRae F. Linton, Richard Sulsky, Jeffrey A. Robl, Rex A. Parker, Goekhan S. Hotamisligil

NATURE ( NATURE PUBLISHING GROUP ) 447 ( 7147 ) 959 - U2 2007.06 [Refereed]

Adipocyte fatty-acid-binding protein, aP2 (FABP4) is expressed in adipocytes and macrophages, and integrates inflammatory and metabolic responses. Studies in aP2-deficient mice have shown that this lipid chaperone has a significant role in several aspects of metabolic syndrome, including type 2 diabetes and atherosclerosis. Here we demonstrate that an orally active small-molecule inhibitor of aP2 is an effective therapeutic agent against severe atherosclerosis and type 2 diabetes in mouse models. In macrophage and adipocyte cell lines with or without aP2, we also show the target specificity of this chemical intervention and its mechanisms of action on metabolic and inflammatory pathways. Our findings demonstrate that targeting aP2 with small-molecule inhibitors is possible and can lead to a new class of powerful therapeutic agents to prevent and treat metabolic diseases such as type 2 diabetes and atherosclerosis.

Coordinated regulation of nutrient and inflammatory responses by STAMP2 is essential for metabolic homeostasis

Kathryn E. Wellen, Raquel Fucho, Margaret F. Gregor, Masato Furuhashi, Carlos Morgan, Torstein Lindstad, Eric Vaillancourt, Cem Z. Gorgun, Fahri Saatcioglu, Gokhan S. Hotamisligil

CELL ( CELL PRESS ) 129 ( 3 ) 537 - 548 2007.05 [Refereed]

Metabolic and inflammatory pathways crosstalk at many levels, and, while required for homeostasis, interaction between these pathways can also lead to metabolic dysregulation under conditions of chronic stress. Thus, we hypothesized that mechanisms might exist to prevent overt inflammatory responses during physiological fluctuations in nutrients or under nutrient-rich conditions, and we identified the six-transmembrane protein STAMP2 as a critical modulator of this integrated response system of inflammation and metabolism in adipocytes. Lack of STAMP2 in adipocytes results in aberrant inflammatory responses to both nutrients and acute inflammatory stimuli. Similarly, in whole animals, visceral adipose tissue of STAMP2(-/-) mice exhibits overt inflammation, and these mice develop spontaneous metabolic disease on a regular diet, manifesting insulin resistance, glucose intolerance, mild hyperglycemia, dyslipidemia, and fatty liver disease. We conclude that STAMP2 participates in integrating inflammatory and metabolic responses and thus plays a key role in systemic metabolic homeostasis.

Chemical chaperones reduce ER stress and restore glucose homeostasis in a mouse model of type 2 diabetes

Umut Oezcan, Erkan Yilmaz, Lale Oezcan, Masato Furuhashi, Eric Vaillancourt, Ross O. Smith, Cem Z. Goerguen, Goekhan S. Hotamisligil

SCIENCE ( AMER ASSOC ADVANCEMENT SCIENCE ) 313 ( 5790 ) 1137 - 1140 2006.08 [Refereed]

Endoplasmic reticulum ( ER) stress is a key link between obesity, insulin resistance, and type 2 diabetes. Here, we provide evidence that this mechanistic link can be exploited for therapeutic purposes with orally active chemical chaperones. 4-Phenyl butyric acid and taurine-conjugated ursodeoxycholic acid alleviated ER stress in cells and whole animals. Treatment of obese and diabetic mice with these compounds resulted in normalization of hyperglycemia, restoration of systemic insulin sensitivity, resolution of fatty liver disease, and enhancement of insulin action in liver, muscle, and adipose tissues. Our results demonstrate that chemical chaperones enhance the adaptive capacity of the ER and act as potent antidiabetic modalities with potential application in the treatment of type 2 diabetes.

Acute renal failure likely due to acute nephritic syndrome associated with typhoid fever

M Hayashi, H Kouzu, M Nishihara, T Takahashi, M Furuhashi, K Sakamoto, N Satoh, T Nishitani, Y Shikano

INTERNAL MEDICINE ( JAPAN SOC INTERNAL MEDICINE ) 44 ( 10 ) 1074 - 1077 2005.10 [Refereed]

A 45-year-old woman was admitted to our hospital because of high fever and malaise. She also presented abnormal urine findings including macrohematuria and proteinuria with transient renal insufficiency. Salmonella Typhi was isolated from her blood and stool culture, and then she was diagnosed as having typhoid fever. Salmonella enterica serotype typhi infection can be complicated by renal involvement, although rare. In Japan, few cases have been reported about acute nephritic syndrome in typhoid fever. Here, we report a case of endemic acquired typhoid fever associated with acute renal failure probably due to acute nephritic syndrome successfully treated with levofloxacin.

Adipocyte/macrophage fatty acid binding proteins control integrated metabolic responses in obesity and diabetes

K Maeda, HM Cao, K Kono, CZ Gorgun, M Furuhashi, KT Uysal, Q Cao, G Atsumi, H Malone, B Krishnan, Y Minokoshi, BB Kahn, RA Parker, GS Hotamisligil

CELL METABOLISM ( CELL PRESS ) 1 ( 2 ) 107 - 119 2005.02 [Refereed]

Fatty acid binding proteins (FABPs) are cytosolic fatty acid chaperones whose biological role and mechanisms of action are not well understood. Here, we developed mice with targeted mutations in two related adipocyte FABPs, aP2 and mal1, to resolve their role in systemic lipid, glucose, and energy metabolism. Mice lacking aP2 and mail exhibited a striking phenotype with strong protection from diet-induced obesity, insulin resistance, type 2 diabetes, and fatty liver disease. These mice have altered cellular and systemic lipid transport and composition, leading to enhanced insulin receptor signaling, enhanced muscle AMP-activated kinase (AMP-K) activity, and dramatically reduced liver stearoyl-CoA desaturase-1 (SCD-1) activity underlying their phenotype. Taken together with the previously reported strong protection against atherosclerosis, these results demonstrate that adipocyte/macrophage FABPs have a robust impact on multiple components of metabolic syndrome, integrating metabolic and inflammatory responses in mice and constituting a powerful target for the treatment of these diseases.

Low adiponectin level in young normotensive men with a family history of essential hypertension

Masato Furuhashi, Nobuyuki Ura, Katsuhiro Higashiura, Yoshinori Miyazaki, Hideyuki Murakami, Masaya Hyakukoku, Kazuaki Shimamoto

HYPERTENSION RESEARCH ( JAPANESE SOC HYPERTENSION CENT ACADEMIC SOC, PUBL OFFICE ) 28 ( 2 ) 141 - 146 2005.02 [Refereed]

Circulating level of adiponectin, an adipocyte-derived protein, is reduced in states of insulin resistance such as obesity and type 2 diabetes. We have previously shown that hypoadiponectinemia is related to insulin resistance in essential hypertension. Recent studies have shown that normotensive subjects with a positive family history of essential hypertension (FH+) have decreased insulin sensitivity compared to subjects with a negative family history of essential hypertension (FH-). We here examined the association between adiponectin concentration and insulin sensitivity in FH+ and FH-. Thirty young, non-obese and normotensive men without a family history of diabetes mellitus were enrolled. A total of 15 subjects were FH+, and the remaining 15 subjects were FH-. Insulin sensitivity index (ISI) was evaluated by the euglycemic hyperin-sulinemic glucose clamp technique. Concentrations of adiponectin and other metabolic variables were measured. The FH+ group had significantly lower levels of ISI and adiponectin than did the FH- group. In all of the subjects, ISI was positively correlated with adiponectin concentration and high-density lipoprotein (HDL) cholesterol level and was negatively correlated with insulin level. Adiponectin concentration was the only independent determinant of ISI in a multiple regression analysis. Our results showed that adiponectin level was significantly decreased and that this was accompanied by reduced insulin sensitivity in young, non-obese and normotensive men with a family history of essential hypertension. Phenotype of reduced adiponectin level as an earlier penetrance may be especially useful in genetic analyses of insulin resistance and essential hypertension.

Liddle's syndrome caused by a novel mutation in the proline-rich PY motif of the epithelial sodium channel beta-subunit

M Furuhashi, K Kitamura, M Adachi, T Miyoshi, N Wakida, N Ura, Y Shikano, Y Shinshi, K Sakamoto, M Hayashi, N Satoh, T Nishitani, K Tomita, K Shimamoto

JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM ( ENDOCRINE SOC ) 90 ( 1 ) 340 - 344 2005.01 [Refereed]

Liddle's syndrome is an autosomal dominant form of salt-sensitive hypertension and has been shown to be caused by missense or frameshift mutations in the amiloride-sensitive epithelial sodium channel (ENaC), which is composed of three subunits: alpha, beta, and gamma. All disease mutations either remove or alter amino acids of the target proline-rich PPPxY sequence (PY motif) of beta- or gamma-ENaC and result in increased channel activity. In this report, we present a family with Liddle's syndrome whose abnormality is caused by a novel missense mutation, P616R, in the PY motif of the betaENaC. Functional studies using the P616R mutant expressed in Xenopus oocytes showed an approximately 6-fold increase in the amiloride-sensitive sodium channel activity compared with that of the wild type. These findings provide additional clinical evidence that a conserved PY motif is critically important for the regulation of ENaC activity.

Blockade of the renin-angiotensin system decreases adipocyte size with improvement in insulin sensitivity

M Furuhashi, N Ura, H Takizawa, D Yoshida, N Moniwa, H Murakami, K Higashiura, K Shimamoto

JOURNAL OF HYPERTENSION ( LIPPINCOTT WILLIAMS & WILKINS ) 22 ( 10 ) 1977 - 1982 2004.10 [Refereed]

Objective Based on results of in vitro studies, it has been hypothesized that blockade of the renin-angiotensin system (RAS) promotes the recruitment and differentiation of pre-adipocytes and that increased formation of small insulin-sensitive adipocytes counteracts ectopic deposition of lipids, thereby improving insulin sensitivity. We investigated the effect of RAS blockade on insulin sensitivity, adipocyte size, and intramuscular lipid content in fructose-fed rats (FFR) as a model of insulin-resistant hypertension. Design and methods Six-week-old male Sprague-Dawley rats were divided into two groups: those fed a standard chow (control) and those fed a fructose-rich chow for 6 weeks. FFR were treated with a vehicle or with 1 mg/kg per day of temocapril, an angiotensin-converting enzyme inhibitor, or 0.1 mg/kg per day of olmesartan, an angiotensin II type 1 receptor blocker, for the last 2 weeks. Insulin sensitivity (M value: mg/kg per min) was estimated by the euglycemic hyperinsulinemic glucose clamp method. Sizes of adipocytes derived from epididymal fat and triglyceride content in the soleus muscle were determined. Results FFR had lower M value, higher blood pressure, larger adipocyte size, higher ratio of epididymal fat pads over body weight (%fat pads), and higher intramuscular triglyceride than did the control rats. Both temocapril and olmesartan significantly improved the M value and decreased blood pressure and adipocyte size without change in %fat pads in FFR. Adipocyte size was negatively correlated with the M value. Treatment for 2 weeks decreased, but not significantly, intramuscular triglyceride. Conclusions RAS blockade decreases adipocyte size without change in epididymal %fat pads accompanied by improvement in insulin sensitivity.

Possible impairment of transcardiac utilization of adiponectin in patients with type 2 diabetes

M Furuhashi, N Ura, N Moniwa, Y Shinshi, H Kouzu, M Nishihara, N Kokubu, T Takahashi, K Sakamoto, M Hayashi, N Satoh, T Nishitani, Y Shikano, K Shimamoto

DIABETES CARE ( AMER DIABETES ASSOC ) 27 ( 9 ) 2217 - 2221 2004.09 [Refereed]

OBJECTIVE - Adiponectin, an adipocyte-derived protein, has been suggested to enhance insulin sensitivity and prevent atherosclerosis. Circulating adiponecin levels are reduced in states of insulin resistance such as type 2 diabetes. We examined transcardiac utilization of adiponectin in patients with and without type 2 diabetes. RESEARCH DESIGN AND METHODS - A total of 17 male type 2 diabetic patients and 17 male nondiabetic patients were investigated. Venous blood samples were taken to measure glucose and lipid variables. Blood samples for the measurement of adiponectin were collected simultaneously from the aortic root and coronary sinus. Angiographic semiquantitative stenosis score of coronary artery was also evaluated. RESULTS - The adiponectin levels in both the aortic root and coronary sinus in the diabetic patients were significantly lower than those in the nondiabetic patients. The adiponectin level was significantly lower in the coronary sinus than in the aortic root in the nondiabetic patients, but there was no significant difference between adiponectin levels in the aortic root and coronary sinus in the diabetic patients. The total stenosis score, as an index of severity of coronary artery stenosis, was significantly higher in the diabetic Patients than in the nondiabetic patients. The stenosis score was correlated with the degree of transcardiac utilization of adiponectin from the aortic root to coronary sinus in the nondiabetic patients but not in the diabetic patients. CONCLUSIONS - Diabetic patients not only have a decreased adiponectin level in the basal state compared with nondiabetic patients but also have impaired utilization of adiponectin in the coronary artery and/or the heart, which may promote the development of atherosclerosis.

Utility of serum ratio of heart-type fatty acid-binding protein to myoglobin for cardiac damage regardless of renal dysfunction

M Furuhashi, N Ura, K Hasegawa, K Tsuchihashi, T Nakata, K Shimamoto

CIRCULATION JOURNAL ( BLACKWELL PUBLISHING ASIA ) 68 ( 7 ) 656 - 659 2004.07 [Refereed]

Background The serum ratio of heart-type fatty acid-binding protein to myoglobin (F/M) has been shown to be a new marker for cardiac damage and volume overload in hemodialysis patients. We evaluated the utility of F/M in hemodialysis patients compared to control subjects. Methods and Results Twenty-one hemodialysis patients and 17 control subjects with normal renal function were investigated. Using a cutoff point of mean + 2SD of the F/M in the controls (value: 0.147), the hemodialysis patients were divided into 2 groups. The concentrations of atrial natriuretic peptide (ANP) and brain natriuretic peptide after hemodialysis in the high F/M group were higher than those in the low F/M group. The mean level of ANP in the low F/M group was almost the upper limit of normal range. The values of left ventricular end-diastolic dimension and left ventricular mass index in the high F/M group were higher than those in the low F/M group, and the mean levels of those in the low F/M group were almost the same as those in the controls. Conclusions The F/M can be evaluated by the absolute value regardless of renal dysfunction, and the value of 0.147 might be useful for determining the cutoff level of cardiac involvement.

血液透析患者におけるアディポネクチンの意義

林学, 古橋眞人, 西谷隆宏, 神津英至, 西原昌宏, 高橋亨, 坂本賢一, 佐藤直利, 鹿野泰邦, 茂庭仁人, 浦信行, 島本和明

日本透析医学会雑誌 ( (一社)日本透析医学会 ) 37 ( Suppl.1 ) 909 - 909 2004.05

血液透析及び腹膜透析患者におけるアディポネクチン濃度

西谷隆宏, 古橋眞人, 林学, 神津英至, 西原昌宏, 高橋亨, 坂本賢一, 佐藤直利, 鹿野泰邦, 茂庭仁人, 浦信行, 島本和明

日本透析医学会雑誌 ( (一社)日本透析医学会 ) 37 ( Suppl.1 ) 796 - 796 2004.05

糖尿病患者におけるアディポネクチン利用障害(Impaired Utilization of Adiponectin in Patients with Diabetes Mellitus)

Furuhashi Masato, Ura Nobuyuki, Takagawa Yoshitoki, Moniwa Norihito, Kouzu Hidemichi, Takahashi Toru, Sakamoto Ken-ichi, Hayashi Manabu, Satoh Naotoshi, Nishitani Takahiro, Shikano Yasukuni, Shinshi Yashuyuki, Kokubu Nobuaki, Shimamoto Kazuaki

Circulation Journal ( (一社)日本循環器学会 ) 68 ( Suppl.I ) 241 - 241 2004.03

Circulating resistin levels in essential hypertension

Masato Furuhashi, Nobuyuki Ura, Katsuhiro Higashiura, Hideyuki Murakami, Kazuaki Shimamoto

CLINICAL ENDOCRINOLOGY ( BLACKWELL PUBLISHING ) 59 ( 4 ) 507 - 510 2003.10 [Refereed]

OBJECTIVE Resistin, a novel cysteine-rich protein secreted by adipocytes, has been proposed to serve as a link between obesity and insulin resistance in rodents, but this has remained controversial. Most of the data obtained in previous studies are restricted to mRNA levels in tissues. DESIGN AND PATIENTS We examined the association between insulin resistance and circulating protein levels of resistin in 33 essential hypertensive patients (EHT) and 18 normotensive subjects (NT). insulin sensitivity (M-value) was evaluated by the euglycaemic hyperinsulinaemic glucose clamp technique. RESULTS Using a cutoff point of mean - 1 SD of the M-value in the NT, the EHT were divided into two groups: one group of 12 insulin-resistant patients (EHT-R) and one group of 21 noninsulin-resistant patients (EHT-N). There were no intergroup differences in age, gender, body mass index (BMI; range: 20.1-30.4 kg/m(2)), fasting glucose and total cholesterol. The EHT-R had significantly higher levels of fasting insulin and triglyceride than did the NT and the EHT-N. The EHT-R had higher levels of free fatty acid and lower levels of high-density lipoprotein cholesterol than did the EHT-N. The M-value was negatively correlated with fasting insulin, free fatty acid, and triglyceride. Circulating resistin levels were not significantly different among the three groups and were not correlated with the M-value, BMI, blood pressure, or lipid variables. CONCLUSIONS Our results suggest that circulating resistin levels are not related to insulin resistance, at least in patients with essential hypertension, although disturbance of lipid metabolism may be associated with insulin resistance.

Role of adiponectin in insulin-resistant hypertension and atherosclerosis

H Murakami, N Ura, M Furuhashi, K Higashiura, T Miura, K Shimamoto

HYPERTENSION RESEARCH ( JAPANESE SOC HYPERTENSION CENT ACADEMIC SOC, PUBL OFFICE ) 26 ( 9 ) 705 - 710 2003.09 [Refereed]

Insulin resistance is one of the major risk factors associated with development of hypertension and atherosclerosis. Recent studies have shown that adiponectin, an adipocyte-derived hormone, may be involved in insulin resistance and development of atherosclerosis in diabetes patients. The aim of this study was to examine adiponectin levels in patients with essential hypertension to determine the relationships between adiponectin levels and insulin sensitivity and to examine the relationship of adiponectin with pulse wave velocity (PWV) in a general population based on the results of an epidemiological survey in Japan. In a clinical study, 20 normotensives (NT) and 30 non-treated essential hypertensives (EHT) were hospitalized, and euglycemic hyperinsulinemic glucose clamp (GC) was performed to evaluate insulin sensitivity defined as M value. EHT were divided into insulin-resistant EHT (EHT-R) and insulin-nonresistant EHT (EHT-N) according to the mean -1 SD of the M value of NT as a cut-off point. Fasting plasma glucose (FPG), immunoreactive insulin (IRI), and adiponectin concentrations were measured. There were no significant differences in body mass index (BMI) or FPG among the NT, EHT-N, and EHT-R groups. The M value and adiponectin concentration in EHT-R were significantly lower than those in the NT or EHT-N. The IRI level in the EHT-R was significantly higher than those in the other groups. A positive correlation between adiponectin concentration and M value was found in all subjects, and adiponectin concentration and M value were found to be significant determinants of each other in multiple regression analysis. In an epidemiological study, we studied 391 male inhabitants of rural communities in Hokkaido, Japan. Systolic blood pressure (SBP), BMI, FPG, IRI, and adiponectin were measured in all subjects early in the morning. Homeostasis model assessment (HOMA) values were calculated as an index of insulin sensitivity, and PWV was used as an index of atherosclerosis. A negative correlation between HOMA values and adiponectin concentration was found in all of the subjects. Multiple regression analysis revealed that adiponectin was a significant determinant for PWV in subjects less than 70 years of age. The results of the clinical study indicate that EHT-R had not only hyperinsulinemia but also a low concentration of adiponectin. The results of multiple regression analysis for determinants of degree of PWV using data obtained in the epidemiological study suggest that adiponectin plays a role in antiatherosclerosis, partly through improvement of insulin resistance.

Right bundle branch block and coved-type ST-segment elevation mimicked by acute cholecystitis

M Furuhashi, K Uno, S Satoh, K Hoshikawa, E Sakai, K Tsuchihashi, K Shimamoto

CIRCULATION JOURNAL ( BLACKWELL PUBLISHING ASIA ) 67 ( 9 ) 802 - 804 2003.09 [Refereed]

A 69-year-old woman had acute cholecystitis that mimicked right bundle branch block with coved-type ST-segment elevation in the precordial electrocardiogram leads (Brugada-type ST shift). The patient did not have obvious heart disease, syncope, or a family history of sudden death. The coved-type ST-segment elevation disappeared as the acute inflammation subsided. Intravenous administration of pilsicainide, a pure sodium channel blocker, could reproduce the Brugada-type ST shift. This is the first report of the Brugada-type ST shift occurring in association with acute cholecystitis.

Blockade of the renin-angiotensin system increases adiponectin concentrations in patients with essential hypertension

M Furuhashi, N Ura, K Higashiura, H Murakami, M Tanaka, N Moniwa, D Yoshida, K Shimamoto

HYPERTENSION ( LIPPINCOTT WILLIAMS & WILKINS ) 42 ( 1 ) 76 - 81 2003.07 [Refereed]

Adiponectin, an adipocyte-derived protein, has been suggested to play an important role in insulin sensitivity. We examined the association between insulin sensitivity (M value) evaluated by the euglycemic-hyperinsulinemic glucose clamp and adiponectin concentrations in 30 essential hypertensives (EHT) and 20 normotensives (NT) and investigated the effect of blockade of the renin-angiotensin system (RAS) on adiponectin concentrations. EHT were divided into 12 insulin-resistant EHT (EHT-R) and 18 non-insulin-resistant EHT (EHT-N) using mean-1 SD of the M value in NT. There were no intergroup differences in age, gender, and body mass index (BMI). EHT-R had significantly higher levels of insulin and triglyceride and lower levels of adiponectin than did NT and EHT-N. EHT-R had higher levels of free fatty acid and lower levels of high-density lipoprotein (HDL) cholesterol than did EHT-N. Adiponectin concentrations were positively correlated with M value and HDL cholesterol and negatively correlated with BMI, insulin, free fatty acid, and triglyceride but not with blood pressure. M value, BMI, and HDL cholesterol were independent determinants of adiponectin concentrations in multiple and stepwise regression analyses. Sixteen EHT were treated with an angiotensin-converting enzyme inhibitor (temocapril, 4 mg/d; n=9) or an angiotensin II receptor blocker (candesartan, 8 mg/d; n=7) for 2 weeks. Treatment with temocapril or candesartan significantly decreased blood pressure and increased M value and adiponectin concentrations but did not affect BMI and HDL cholesterol. These results suggest that hypoadiponectinemia is related to insulin resistance in essential hypertension and that RAS blockade increases adiponectin concentrations with improvement in insulin sensitivity.

The effect of tumor necrosis factor-alpha on tissue specificity and selectivity to insulin signaling

K Yamaguchi, K Higashiura, N Ura, H Murakami, M Hyakukoku, M Furuhashi, K Shimamoto

HYPERTENSION RESEARCH ( JAPANESE SOC HYPERTENSION CENT ACADEMIC SOC, PUBL OFFICE ) 26 ( 5 ) 389 - 396 2003.05 [Refereed]

Recent studies have indicated that tumor necrosis factor (TNF)-alpha plays a significant role in insulin resistance. It has been proposed that selective impairment of insulin signaling in glucose metabolism is related to the development of atherosclerosis, although the mechanisms are not clear. The aim of this study was to elucidate the effect of TNF-alpha on tissue specificity and selectivity to insulin signaling. L6 myotubes and rat aortic vascular smooth muscle cells (VSMC) were cultured. Cells were stimulated with insulin pretreated with or without TNF-alpha. The protein extracts were used for electrophoresis and immunoblotting studies to examine phosphorylation of insulin receptor (IR)-beta, insulin receptor substrate (IRS)-1 and extracellular signal-regulated kinase (ERK). IR-beta phosphorylation was not affected by TNF-alpha in L6 or in VSMC. TNF-alpha significantly (p<0.05) inhibited IRS-1 phosphorylation by insulin but had no effect on ERK in L6. TNF-alpha had no effect on either IRS-1 phosphorylation or ERK in VSMC. Insulin induced ERK phosphorylation in a dose-dependent manner in VSMC. These results suggests that TNF-alpha plays a significant role in the tissue specificity and signal selectivity of insulin resistance. The pathway related to glucose metabolism is selectively impaired by TNF-alpha in skeletal muscle, and this impairment may induce compensatory hyperinsulinemia, which in turn would stimulate the pathway related to the cell proliferation in vascular tissues and possibly enhance the progression of atherosclerosis.

Tissue-specific impairment of insulin signaling in vasculature and skeletal muscle of fructose-fed rats

M Hyakukoku, K Higashiura, N Ura, H Murakami, K Yamaguchi, L Wang, M Furuhashi, N Togashi, K Shimamoto

HYPERTENSION RESEARCH ( JAPANESE SOC HYPERTENSION CENT ACADEMIC SOC, PUBL OFFICE ) 26 ( 2 ) 169 - 176 2003.02 [Refereed]

The relation between insulin resistance/hyperinsulinemia and cardiovascular diseases has attracted much attention. Insulin affects not only glucose metabolism, but also protein synthesis and cell growth. Insulin stimulates both the phosphatidylinositol 3-kinase (PI3-K) and mitogen-activated protein kinase (MAPK) pathways, but the relationship between cardiovascular disease and selective insulin signal pathways is unclear. We investigated the tissue specificity and intracellular signal transduction selectivity of insulin resistance in the vasculature and skeletal muscle of fructose-fed rats (FFR). Sprague-Dawley rats were fed either normal rat chow (control rats) or fructose-rich chow. Normal saline with or without 1,000 (mug/kg) insulin was injected, and then the thoracic aorta or soleus muscle was removed under anesthetization. Insulin-induced tyrosine phosphorylation of insulin receptor beta subunit (IRbeta) and insulin receptor substrate-1 (IRS-1) and tyrosine/threonine phosphorylation of p44/42 MAPK (ERK-1/2) were evaluated. There were no significant differences in the degree of phosphorylation of IRbeta or ERK-1/2 in the thoracic aorta or in the soleus muscle between FFR and controls. However, tyrosine phosphorylation of IRS-1 in the soleus muscle of FFR was significantly reduced to 80% (p<0.001) of that in controls. The results suggest that PI3-K pathway in skeletal muscle is selectively impaired in FFR, and this impairment may induce hyperinsulinemia, which in turn may stimulate the MAPK pathway and lead to atherosclerosis. Thus PI3-K pathway may be one of the factors underlying the onset of cardiovascular disease in patients with insulin resistance.

Insulin sensitivity and lipid metabolism in human CD36 deficiency

M Furuhashi, N Ura, T Nakata, K Shimamoto

DIABETES CARE ( AMER DIABETES ASSOC ) 26 ( 2 ) 471 - 474 2003.02 [Refereed]

OBJECTIVE - CD36 has been proposed as a fatty acid translocase and a receptor for HDL and oxidized LDL. The association between CD36 deficiency and insulin resistance remains controversial. We investigated glucose and lipid metabolism in human CD36 deficiency. RESEARCH DESIGN AND METHODS - A total of 61 type I CD36-deficient patients and 25 control subjects were examined. Diabetes was defined as fasting glucose level greater than or equal to7 mmol/l or use of hypoglycemic agents. A homeostasis model assessment (HOMA) index was evaluated in patients without diabetes. Insulin resistance was defined as a HOMA index greater than or equal to1.73 (sensitivity 64.3%, specificity 78.9%; J Japan Diab Soc, 2000). RESULTS - Diabetes was identified in 12 (20%) of the 61 CD36-deficient patients. Fasting glucose, HbA(1c), and total cholesterol levels in the diabetic CD36-deficient patients were significantly higher than in the control subjects and the nondiabetic CD36-deficient patients. Regardless of diabetes, HDL cholesterol concentrations in the CD36-deficient patients were significantly higher than in the control subjects. The nondiabetic CD36-deficient patients had higher triglyceride concentrations than the control subjects, and triglyceride concentrations were higher in the diabetic CD36-deficient patients than in the nondiabetic CD36-deficient patients. The prevalence of insulin resistance in the nondiabetic CD36-deficient patients was similar to that in the control subjects. CONCLUSIONS - Human CD36 deficiency is not necessarily responsible for insulin resistance. Lipid abnormalities in CD36 deficiency may partly depend on the presence of diabetes, and increased levels of triglyceride and HDL cholesterol may be due to impaired binding of fatty acids and HDL to CD36 and subsequent clearance.

Serum ratio of heart-type fatty acid-binding protein to myoglobin - A novel marker of cardiac damage and volume overload in hemodialysis patients

M Furuhashi, N Ura, K Hasegawa, H Yoshida, K Tsuchihashi, T Nakata, K Shimamoto

NEPHRON CLINICAL PRACTICE ( KARGER ) 93 ( 2 ) C69 - C74 2003.02 [Refereed]

Background: The concentration of heart-type fatty acid-binding protein (hFABP), a promising novel marker for detection of acute or persistent myocardial damage, is significantly influenced by renal clearance and thus has limitations to its usefulness in patients with renal dysfunction. We evaluated whether the serum ratio of hFABP to myoglobin (F/M) might be a useful marker for assessing cardiac damage in hemodialysis patients. Methods: Serum hFABP and myoglobin were measured, and the value of F/M was calculated in 21 hemodialysis patients. Cardiac markers (cardiac troponin T [cTnT], atrial natriuretic peptide [ANP], and brain natriuretic peptide [BNP]) and echocardiographic indices (left ventricular end-diastolic dimension [LVDd], left ventricular mass index [LVMI], and inferior vena cava [IVC] dimension) were examined and compared with hFABP, myoglobin, and F/M ratios. Results: Serum hFABP and myoglobin levels were significantly elevated in hemodialysis patients and reduced by 30-40% during hemodialysis. The value of F/M after hemodialysis, but not the concentration of hFABP or myoglobin, had significant linear correlations with ANP, BNP, cTnT, LVDd, LVMI, and IVC. Conclusion: The value of F/M after hemodialysis, but not the concentration of hFABP itself, might be a newly useful marker for estimation of cardiac damage and volume overload in hemodialysis patients. Copyright (C) 2003 S. Karger AG, Basel.

Serum ratio of heart-type fatty acid-binding protein to myoglobin. A novel marker of cardiac damage and volume overload in hemodialysis patients.

Furuhashi M, Ura N, Hasegawa K, Yoshida H, Tsuchihashi K, Nakata T, Shimamoto K

Nephron. Clinical practice ( 2 ) 93 C69 - 74 2003 [Refereed]

本態性高血圧におけるレジスチンとインスリン抵抗性

古橋眞人, 浦信行, 吉田大輔, 田中希尚, 茂庭仁人, 萩原誠, 村上英之, 吉田英昭, 東浦勝浩, 島本和明

日本内分泌学会雑誌 ( (一社)日本内分泌学会 ) 78 ( 2 ) 459 - 459 2002.09

Sonoclot coagulation analysis: new bedside monitoring for determination of the appropriate heparin dose during haemodialysis

M Furuhashi, N Ura, K Hasegawa, H Yoshida, K Tsuchihashi, T Miura, K Shimamoto

NEPHROLOGY DIALYSIS TRANSPLANTATION ( OXFORD UNIV PRESS ) 17 ( 8 ) 1457 - 1462 2002.08 [Refereed]

Background. Thrombotic and haemorrhagic complications affect cardiovascular morbidity and mortality in haemodialysis patients. We investigated whether a new bedside Sonoclot analysis is useful for determining an appropriate heparin dose during haemodialysis. Methods. Twenty-two haemodialysis patients (15 males and seven females) treated with unfractionated heparin were recruited. Sonoclot parameters, including activated clotting time (ACT), clot rate (CR), time to peak (TP) and peak amplitude (PA), were examined before and after haemodialysis, coupled with conventional coagulable variables, platelet count and fibrinogen level. Some indices were determined and verified to control heparin dose. Thrombotic and haemorrhagic complications were observed for 3 months. Sonoclot analysis was subsequently performed before initiation, 1 h later, 2.5 or 3 h later (before cessation of heparin infusion), and at the end of haemodialysis. Results. Both CR and PA were positively correlated with platelet count and fibrinogen level. Sonoclot parameters except for PA were significantly correlated with heparin dose. Heparin dose was reduced without causing complications in 12 patients with DeltaACT greater than or equal to40 s and/or CR after haemodialysis (CRpost) <20 U/min. Thrombotic complications occurred in five patients who had CRpost >30 U/min. ACT and TP increased during heparin treatment. ACT was reversed but TP did not change after cessation of heparin. CR significantly decreased after initiation of heparin and was reversed, although not completely, after cessation of heparin. PA showed no significant changes during haemodialysis. Conclusions. Sonoclot analysis can monitor accurate coagulable states and determine an appropriate heparin dose during haemodialysis.

Fenofibrate improves insulin sensitivity in connection with intramuscular lipid content, muscle fatty acid-binding protein, and beta-oxidation in skeletal muscle

M Furuhashi, N Ura, H Murakami, M Hyakukoku, K Yamaguchi, K Higashiura, K Shimamoto

JOURNAL OF ENDOCRINOLOGY ( SOC ENDOCRINOLOGY ) 174 ( 2 ) 321 - 329 2002.08 [Refereed]

We investigated the effect of fenofibrate, a peroxisome proliferator-activated receptor-alpha agonist, on insulin sensitivity including lipid metabolism in skeletal muscle. Six-week-old male Sprague-Dawley rats were divided into two groups: those fed a standard chow (control) or a fructose-rich chow (fructose-fed rats (FFRs)) for 6 weeks. FFRs were treated either with a vehicle or with 30 mg/kg per day of fenofibrate for the last 2 weeks. Insulin sensitivity (NI-value) was estimated by the euglycemic hyperinsulinernic glucose clamp method. Fatty acid-binding protein (FABP) in skeletal muscle was measured by ELISA, and the expression of FABP mRNA was analyzed by semi-quantitative RT-PCR. The serum and muscle triglyceride (sTG and mTG) levels and the activity of 3-hydroxyacyl-CoA dehydrogenase (HADH), a beta-oxidation enzyme, in muscle were also determined. FFRs showed a lower NI-value and higher blood pressure, sTG and mTG than did the control group. The mTG was correlated positively with sTG and negatively with the NI-value. Fenofibrate treatment for 2 weeks did not change blood pressure but significantly improved the M-value, sTG and mTG. FABP content and mRNA in the soleus muscle were significantly elevated in FFRs compared with those in the control group. Fenofibrate treatment further increased FABP. The HADH activity was comparable between the control group and FFRs, but significantly increased by fenofibrate treatment. These results suggest that fenofibrate improves insulin sensitivity not only by lowering serum lipids and subsequent influx of fatty acids into muscles but also by reducing intramuscular lipid content via further induction of FABP and stimulation of beta-oxidation in muscles.

Prevalence of asymptomatic ST segment elevation in right precordial leads with right bundle branch block (Brugada-type ST shift) among the general Japanese population

M Furuhashi, K Uno, K Tsuchihashi, D Nagahara, M Hyakukoku, T Ohtomo, S Satoh, T Nishimiya, K Shimamoto

HEART ( BRITISH MED JOURNAL PUBL GROUP ) 86 ( 2 ) 161 - 166 2001.08 [Refereed]

Objective-To examine the modality and morbidity of asymptomatic ST segment elevation in leads V1 to V3 with right bundle branch block (Brugada-type ST shift). Methods-8612 Japanese subjects (5987 men and 2625 women, mean age 49.2 years) who underwent a health check up in 1997 were investigated. Those with Brugada-type ST shift underwent the following further examinations over a two year period after the initial check up: ECG, echocardiogram, 24 hour Holter monitoring, treadmill exercise testing, signal averaged ECG, and slow kinetic sodium channel blocker loading test (cibenzoline, 1.4 mg/kg). Results-Asymptomatic Brugada-type ST sh-ift was found in 12 of 8612 (0.14%) subjects. Eleven of these 12 subjects were followed up. Follow up ECG exhibited persistent Brugada-type ST shift in seven of 11 (63.6%) subjects. ST shift was transformed from a saddle back to a coved type in three subjects. None of the subjects had morphological abnormalities or abnormal tachyarrhythmias. Positive late potentials were found in seven of 11 (63.6%) subjects. Augmentation of ST shift was shown by both submaximal exercise and drug administration in one of the 11 subjects (9.1%). Conclusions-Asymptomatic subjects with Brugada-type ST shift were not unusual, at a rate of 0.14% in the general Japanese population. Almost all of the subjects had some abnormalities in non-invasive secondary examinations. Additional and prospective studies are needed to confirm the clinical significance and the prognosis of asymptomatic Brugada-type ST shift.

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