論文 - 鈴木　拓

多発性骨髄腫における新規エピジェネティック併用療法の開発(Development of a new combinational epigenetic therapy of multiple myeloma)

石黒 一也, 北嶋 洋志, 新沼 猛, 石田 禎夫, 丸山 玲緒, 池田 博, 山本 英一郎, 甲斐 正広, 佐々木 泰史, 時野 隆至, 仲瀬 裕志, 鈴木 拓

日本癌学会総会記事 ( (一社)日本癌学会 )  78回   P - 2241  2019年09月

非乳頭十二指腸腫瘍におけるDNAメチル化解析(Methylation analysis of non-ampullary duodenal precancerous and cancerous lesions)

澤田 武, 太田 亮介, 鈴木 拓, 津山 翔, 八尾 隆史, 中西 宏佳, 山本 英一郎, 久保田 英嗣, 片岡 洋望, 佐々木 泰史, 源 利成

日本癌学会総会記事 ( 日本癌学会 )  78回   P - 2036  2019年09月

頭頸部扁平上皮がんの微小環境におけるAEBP1の解析(Analysis of AEBP1 in the microenvironment of head and neck squamous cell carcinoma)

萬 顕, 山本 英一郎, 須藤 豪太, 沼田 有斗, 新沼 猛, 北嶋 洋志, 甲斐 正広, 小島 隆, 高野 賢一, 鈴木 拓

日本癌学会総会記事 ( 日本癌学会 )  78回   P - 1171  2019年09月

正常大腸粘膜のヒストン修飾異常とCIMP大腸腫瘍の発がんリスクの関連(Repressive histone mark in normal colon is associated with the risk of CRC with CpG island methylator phenotype)

山本 英一郎, 須藤 豪太, 久保 俊之, 萬 顕, 原田 拓, 青木 敬則, 北嶋 洋志, 新沼 猛, 甲斐 正広, 菅井 有, 仲瀬 裕志, 鈴木 拓

日本癌学会総会記事 ( 日本癌学会 )  78回   J - 2035  2019年09月

新規大腸がん線維芽細胞関連遺伝子の同定(Identification of a novel cancer associated fibroblast-related gene in colorectal cancer)

須藤 豪太, 山本 英一郎, 萬 顕, 新沼 猛, 杉本 亮, 北嶋 洋志, 久保 俊之, 畠中 柚衣, 甲斐 正広, 時野 隆至, 仲瀬 裕志, 菅井 有, 鈴木 拓

日本癌学会総会記事 ( 日本癌学会 )  78回   P - 3102  2019年09月

UHRF1 depletion and HDAC inhibition reactivate epigenetically silenced genes in colorectal cancer cells.

Takeshi Niinuma, Hiroshi Kitajima, Masahiro Kai, Eiichiro Yamamoto, Akira Yorozu, Kazuya Ishiguro, Hajime Sasaki, Gota Sudo, Mutsumi Toyota, Tomo Hatahira, Reo Maruyama, Takashi Tokino, Hiroshi Nakase, Tamotsu Sugai, Hiromu Suzuki

Clinical epigenetics   11 ( 1 ) 70 - 70  2019年05月  [査読有り]  [国際誌]

　概要を見る

BACKGROUND: Ubiquitin-like protein containing PHD and RING finger domains 1 (UHRF1) is a major regulator of epigenetic mechanisms and is overexpressed in various human malignancies. In this study, we examined the involvement of UHRF1 in aberrant DNA methylation and gene silencing in colorectal cancer (CRC). RESULTS: CRC cell lines were transiently transfected with siRNAs targeting UHRF1, after which DNA methylation was analyzed using dot blots, bisulfite pyrosequencing, and Infinium HumanMethylation450 BeadChip assays. Gene expression was analyzed using RT-PCR and gene expression microarrays. Depletion of UHRF1 rapidly induced genome-wide DNA demethylation in CRC cells. Infinium BeadChip assays and bisulfite pyrosequencing revealed significant demethylation across entire genomic regions, including CpG islands, gene bodies, intergenic regions, and repetitive elements. Despite the substantial demethylation, however, UHRF1 depletion only minimally reversed CpG island hypermethylation-associated gene silencing. By contrast, the combination of UHRF1 depletion and histone deacetylase (HDAC) inhibition reactivated the silenced genes and strongly suppressed CRC cell proliferation. The combination of UHRF1 depletion and HDAC inhibition also induced marked changes in the gene expression profiles such that cell cycle-related genes were strikingly downregulated. CONCLUSIONS: Our results suggest that (i) maintenance of DNA methylation in CRC cells is highly dependent on UHRF1; (ii) UHRF1 depletion rapidly induces DNA demethylation, though it is insufficient to fully reactivate the silenced genes; and (iii) dual targeting of UHRF1 and HDAC may be an effective new therapeutic strategy.

DOI PubMed

DOT1L inhibition blocks multiple myeloma cell proliferation by suppressing IRF4-MYC signaling.

Kazuya Ishiguro, Hiroshi Kitajima, Takeshi Niinuma, Tadao Ishida, Reo Maruyama, Hiroshi Ikeda, Toshiaki Hayashi, Hajime Sasaki, Hideki Wakasugi, Koyo Nishiyama, Tetsuya Shindo, Eiichiro Yamamoto, Masahiro Kai, Yasushi Sasaki, Takashi Tokino, Hiroshi Nakase, Hiromu Suzuki

Haematologica   104 ( 1 ) 155 - 165  2019年01月  [査読有り]  [国際誌]

　概要を見る

Epigenetic alterations play an important role in the pathogenesis in multiple myeloma, but their biological and clinical relevance is not fully understood. Here, we show that DOT1L, which catalyzes methylation of histone H3 lysine 79, is required for myeloma cell survival. DOT1L expression levels were higher in monoclonal gammopathy of undetermined significance and smoldering multiple myeloma than in normal plasma cells. Treatment with a DOT1L inhibitor induced cell cycle arrest and apoptosis in myeloma cells, and strongly suppressed cell proliferation in vitro The anti-myeloma effect of DOT1L inhibition was confirmed in a mouse xenograft model. Chromatin immunoprecipitation-sequencing and microarray analysis revealed that DOT1L inhibition downregulated histone H3 lysine 79 dimethylation and expression of IRF4-MYC signaling genes in myeloma cells. In addition, DOT1L inhibition upregulated genes associated with immune responses and interferon signaling. Myeloma cells with histone modifier mutations or lower IRF4/MYC expression were less sensitive to DOT1L inhibition, but with prolonged treatment, anti-proliferative effects were achieved in these cells. Our data suggest that DOT1L plays an essential role in the development of multiple myeloma and that DOT1L inhibition may provide new therapies for myeloma treatment.

DOI PubMed

Expression of Epithelial-Mesenchymal Transition Proteins in Pancreatic Anaplastic (Undifferentiated) Carcinoma.

Ishida K, Yamashita R, Osakabe M, Uesugi N, Yamada N, Nitta H, Fujishima F, Motoi F, Suzuki H, Shimamura H, Noda Y, Sawai T, Unno M, Sasano H, Sasaki A, Sugai T

Pancreas   48 ( 1 ) 36 - 42  2019年01月  [査読有り]  [国際誌]

　概要を見る

OBJECTIVES: The aim of this study was to identify an association of pancreatic anaplastic carcinoma (APC) with the epithelial-mesenchymal transition (EMT). METHODS: Resected APCs (n = 24) were examined to assess components of APCs, including carcinomatous, transitional, and sarcomatous regions. Analysis was performed based on the immunoreactivity of E-cadherin and 3 EMT-related proteins: Slug (zinc finger protein SNAI2), Twist (Twist-related protein 1), and Zeb1 (zinc finger E-box-binding homeobox 1). Expression score was determined based on staining intensity and stained area of the target cells. Finally, we performed a hierarchical clustering based on the expression pattern of E-cadherin and EMT-related proteins of the sarcomatous component. RESULTS: The expression score of E-cadherin decreased in the order of sarcomatous > transitional > carcinomatous components (P < 0.01). Although there were significant differences in the immunohistochemical scores of Slug, Twist, and Zeb1 between carcinomatous and transitional components (P < 0.01), the significant difference in immunohistochemical score of Zeb1 between transitional and sarcomatous components was found (P < 0.05). Furthermore, APCs were divided into 2 subgroups based on the expression patterns of E-cadherin and EMT-related proteins (hierarchical clustering analysis). Consequently, these subgroups were distinguished by Twist expression. CONCLUSIONS: Epithelial-mesenchymal transition plays an essential role in the pathogenesis of APC.

DOI PubMed

Integrated Analysis of the Endoscopic, Pathological and Molecular Characteristics of Colorectal Tumorigenesis.

Suzuki H, Yamamoto E, Yamano HO, Nakase H, Sugai T

Digestion   99 ( 1 ) 33 - 38  2019年  [査読有り]

DOI PubMed

Molecular biology of colorectal cancer -mechanism of tumorigenesis and molecular subtypes-

Hiromu Suzuki, Eiichiro Yamamoto, Hiroshi Nakase

Journal of Japanese Society of Gastroenterology ( Japanese Society of Gastroenterology )  116 ( 11 ) 859 - 866  2019年

DOI PubMed

Surface microstructures are associated with mutational intratumoral heterogeneity in colorectal tumors.

Taku Harada, Eiichiro Yamamoto, Hiro-O Yamano, Hironori Aoki, Hiro-O Matsushita, Kenjiro Yoshikawa, Ryo Takagi, Eiji Harada, Yoshihito Tanaka, Yuko Yoshida, Makoto Eizuka, Akira Yorozu, Gota Sudo, Hiroshi Kitajima, Takeshi Niinuma, Masahiro Kai, Yasushi Sasaki, Takashi Tokino, Tamotsu Sugai, Hiroshi Nakase, Hiromu Suzuki

Journal of gastroenterology   53 ( 12 ) 1241 - 1252  2018年12月  [査読有り]  [国内誌]

　概要を見る

BACKGROUND: Recent studies revealed that colorectal tumors are composed of genetically diverse subclones. We aimed to clarify whether the surface microstructures of colorectal tumors are associated with genetic intratumoral heterogeneity (ITH). METHODS: The surface microstructures (pit patterns) of colorectal tumors were observed using magnifying endoscopy, and biopsy specimens were obtained from respective areas when tumors exhibited multiple pit patterns. A total of 711 specimens from 477 colorectal tumors were analyzed for BRAF, KRAS and TP53 mutations using pyrosequencing and direct sequencing. A panel of cancer-related genes was analyzed through targeted sequencing in 7 tumors. RESULTS: Colorectal tumors with multiple pit patterns exhibited more advanced pit patterns and higher frequencies of KRAS and/or TP53 mutations than tumors with a single pit pattern. In tumors with multiple pit patterns, mutations were observed as public (common to all areas) or private (specific to certain areas), and private KRAS and/or TP53 mutations were often variable and unrelated to the pit pattern grade. Notably, invasive CRCs frequently exhibited public TP53 mutations, even in adenomatous areas, which is indicative of their early malignant potential. Targeted sequencing revealed additional public and private mutations in tumors with multiple pit patterns, indicating their single clonal origin. CONCLUSIONS: Our results suggest intratumoral pit pattern variation does not simply reflect the process of colorectal tumor evolution, but instead represents genetically diverse subclones, and this diversity may be associated with malignant potential.

DOI PubMed

Dysregulation of miRNA in chronic hepatitis B is associated with hepatocellular carcinoma risk after nucleos(t)ide analogue treatment.

Hideki Wakasugi, Hideaki Takahashi, Takeshi Niinuma, Hiroshi Kitajima, Ritsuko Oikawa, Naoki Matsumoto, Yuko Takeba, Takehito Otsubo, Masayuki Takagi, Yasushi Ariizumi, Michihiro Suzuki, Chiaki Okuse, Shogo Iwabuchi, Masayuki Nakano, Noriyuki Akutsu, Jong-Hon Kang, Takeshi Matsui, Norie Yamada, Hajime Sasaki, Eiichiro Yamamoto, Masahiro Kai, Yasushi Sasaki, Shigeru Sasaki, Yasuhito Tanaka, Hiroshi Yotsuyanagi, Takeya Tsutsumi, Hiroyuki Yamamoto, Takashi Tokino, Hiroshi Nakase, Hiromu Suzuki, Fumio Itoh

Cancer letters   434   91 - 100  2018年10月  [査読有り]  [国際誌]

　概要を見る

Hepatitis B virus (HBV) infection is a major cause of hepatocellular carcinoma (HCC). Nucleos(t)ide analogue (NA) therapy effectively reduces the incidence of HCC, but it does not completely prevent the disease. Here, we show that dysregulation of microRNAs (miRNAs) is involved in post-NA HCC development. We divided chronic hepatitis B (CHB) patients who received NA therapy into two groups: 1) those who did not develop HCC during the follow-up period after NA therapy (no-HCC group) and 2) those who did (HCC group). miRNA expression profiles were significantly altered in CHB tissues as compared to normal liver, and the HCC group showed greater alteration than the no-HCC group. NA treatment restored the miRNA expression profiles to near-normal in the no-HCC group, but it was less effective in the HCC group. A number of miRNAs implicated in HCC, including miR-101, miR-140, miR-152, miR-199a-3p, and let-7g, were downregulated in CHB. Moreover, we identified CDK7 and TACC2 as novel target genes of miR-199a-3p. Our results suggest that altered miRNA expression in CHB contributes to HCC development, and that improvement of miRNA expression after NA treatment is associated with reduced HCC risk.

DOI PubMed

Molecular Profiling Based on KRAS/BRAF Mutation, Methylation, and Microsatellite Statuses in Serrated Lesions.

Sugai T, Eizuka M, Fujita Y, Kawasaki K, Yamamoto E, Ishida K, Yamano H, Suzuki H, Matsumoto T

Digestive diseases and sciences   63 ( 10 ) 2626 - 2638  2018年10月  [査読有り]  [国際誌]

　概要を見る

AIM: The aim of your study is to characterize serrated lesions according to their molecular patterns, specifically BRAF/KRAS mutation, methylation, and microsatellite statuses. We evaluated the molecular patterns of 163 serrated lesions, including 37 microvesicular hyperplastic polyps, 73 sessile serrated adenomas/polyps (SSA/Ps), 31 traditional serrated adenomas, and 22 SSA/Ps with cytological dysplasia/adenocarcinoma. METHODS: Mutations in BRAF (V600E)/KRAS (exon 2) and microsatellite status [microsatellite stability (MSS) vs. MSI] were examined using a pyrosequencer and the PCR-based microsatellite method, respectively. DNA methylation status was classified as low (LME), intermediate (IME), or high methylation epigenotype (HME) according to a PCR-based two-step method. In addition, mucin and annexin A10 expression was examined. Finally, we performed a hierarchical clustering analysis of the BRAF/KRAS mutation, DNA methylation, and microsatellite statuses. RESULTS: The molecular patterns observed in the serrated lesions could be divided into five subgroups: lesions characterized by (1) BRAF mutation, HME, and MSI; (2) BRAF mutation, HME, and MSS; (3) BRAF mutation, LME/IME, and MSS; (4) no BRAF/KRAS mutations, LME/IME, and MSS; and (5) KRAS mutation, LME/IME, and MSS. In addition, we demonstrated that these observed molecular patterns help identify the associations of the molecular patterns and markers (i.e., mucin and annexin A10) with the clinicopathological findings, including histological features and histological diagnosis. CONCLUSIONS: We suggest that the identified molecular patterns play an important role in the pathway of serrated lesion development.

DOI PubMed

MicroRNA-31 reflects IL-6 expression in cancer tissue and is related with poor prognosis in bile duct cancer.

Keisuke Ishigami, Katsuhiko Nosho, Hideyuki Koide, Shinichi Kanno, Kei Mitsuhashi, Hisayoshi Igarashi, Masahiro Shitani, Masayo Motoya, Yasutoshi Kimura, Tadashi Hasegawa, Hiroyuki Kaneto, Ichiro Takemasa, Hiromu Suzuki, Hiroshi Nakase

Carcinogenesis   39 ( 9 ) 1127 - 1134  2018年09月  [査読有り]  [国際誌]

　概要を見る

Bile duct cancer is a highly aggressive malignancy wherein early diagnosis is difficult and few treatment options are available. MicroRNA-31 (miR-31) is reported to be related with survival in patients with gastrointestinal cancers; however, the regulatory mechanism of miR-31 and association between miR-31 expression and survival in patients with bile duct cancer cases have not been established. Thus, we evaluated miR-31 expression in bile duct cancer tissues and assessed its relationship with prognosis. Additionally, we examined the effects of several cytokines on miR-31 expression. The study included 81 samples of bile duct cancer tissues. MiR-31 expression in bile duct cancer cells was significantly higher than that in normal bile duct epithelial cells (P = 0.038). There were no significant associations between miR-31 expression and clinical or pathological characteristics, except for tumour size (P = 0.012). In Kaplan-Meier analysis, high miR-31 expression was significantly associated with shorter survival (log-rank test, P = 0.0082). In multivariate Cox regression analysis, high miR-31 expression was significantly associated with prognosis (P = 0.043), independent of clinical or pathological features. Interleukin-6 (IL-6) significantly promoted miR-31 expression and cell proliferation in a dose-dependent manner, and the inhibition of STAT-3 signalling significantly suppressed miR-31 expression and cell proliferation. In conclusion, high expression was significantly associated with poor prognosis in bile duct cancer patients. The IL-6-STAT-3 signalling regulated bile duct cancer cell proliferation and miR-31 expression. Our findings suggest that miR-31 may be a promising biomarker that reflects IL-6 expression in bile duct cancer tissues and predicts poor prognosis.

DOI PubMed

新規大腸がん線維芽細胞関連遺伝子の同定(Identification of cancer-associated fibroblast-related genes in colorectal cancer)

沼田 有斗, 山本 英一郎, 萬 顕, 新沼 猛, 杉本 亮, 北嶋 洋志, 甲斐 正広, 青木 敬則, 須藤 豪太, 時野 隆至, 仲瀬 裕志, 菅井 有, 鈴木 拓

日本癌学会総会記事 ( 日本癌学会 )  77回   2189 - 2189  2018年09月

大腸鋸歯状病変における、遺伝子変異、コピー数変化、DNAメチル化の統合解析(Integrative analysis of gene mutations, copy number alterations and DNA methylation in colorectal serrated lesions)

澤田 武, 中西 宏佳, 佐々木 泰史, 山本 英一郎, 青木 敬則, 永塚 真, 高橋 直樹, 太田 亮介, 久保田 英嗣, 片岡 洋望, 源 利成, 菅井 有, 鈴木 拓

日本癌学会総会記事 ( (一社)日本癌学会 )  77回   1872 - 1872  2018年09月

SMOC1のエピジェネティックなサイレンシングは大腸鋸歯状腺腫の発育進展に関与する(Epigenetic silencing of SMOC1 is associated with development of colorectal traditional serrated adenomas)

青木 敬則, 山本 英一郎, 高澤 啓, 新沼 猛, 山野 泰穂, 萬 顕, 北嶋 洋志, 甲斐 正広, 澤田 典均, 仲瀬 裕志, 菅井 有, 鈴木 拓

日本癌学会総会記事 ( 日本癌学会 )  77回   1871 - 1871  2018年09月

肝細胞癌におけるBET阻害剤の抗腫瘍メカニズムの解析(Analysis of antitumor mechanism of BET inhibition in hepatocellular carcinoma)

佐々木 基, 新沼 猛, 北嶋 洋志, 山本 英一郎, 石黒 一也, 若杉 英樹, 萬 顕, 須藤 豪太, 甲斐 正広, 鈴木 拓, 仲瀬 裕志

日本癌学会総会記事 ( 日本癌学会 )  77回   1387 - 1387  2018年09月

大腸がんにおける腫瘍血管内皮関連遺伝子の同定(Identification and characterization of a tumor endothelium-related gene in colorectal cancer)

萬 顕, 山本 英一郎, 沼田 有斗, 新沼 猛, 北嶋 洋志, 甲斐 正広, 須藤 豪太, 黒瀬 誠, 時野 隆至, 仲瀬 裕志, 菅井 有, 鈴木 拓

日本癌学会総会記事 ( 日本癌学会 )  77回   1574 - 1574  2018年09月

トリプルネガティブ乳癌におけるSALL3の高頻度なエピジェネティックな不活化機構(Downregulation of SALL3 by recurrent genetic and epigenetic alterations is involved in triple negative breast cancers)

松下 洋輔, 小松 正人, 清谷 一馬, 吉丸 哲郎, 新沼 猛, 鈴木 拓, 本田 純子, 井本 逸勢, 丹黒 章, 三好 康雄, 笹 三徳, 片桐 豊雅

日本癌学会総会記事 ( 日本癌学会 )  77回   1410 - 1410  2018年09月  [査読有り]