Affiliation 【 display / non-display 】

Affiliation 【 display / non-display 】

• Sapporo Medical University   Department of Pathology   Assistant Professor  

Papers 【 display / non-display 】

Papers 【 display / non-display 】

• An SNP-dependent cancer-testis antigenic epitope serves as a promising immunotherapeutic target for cancer.

  Kenji Murata, Tomoyuki Minowa, Tomohide Tsukahara, Taku Yoshida, Akiko Minami, Munehide Nakatsugawa, Yuka Mizue, Aiko Murai, Serina Tokita, Kenta Sasaki, Hisashi Uhara, Terufumi Kubo, Takayuki Kanaseki, Toshihiko Torigoe, Yoshihiko Hirohashi

  Oncoimmunology   14 ( 1 ) 2528110 - 2528110  2025.12  [International journal]

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  T cells recognize peptides presented by human leukocyte antigen molecules on the cell surface, enabling the immune surveillance of pathological conditions such as cancer. Cancer-testis (CT) antigens are promising targets for cancer immunotherapy because of their restricted expression in normal tissues. In this study, we performed antigen screening of T cell receptors isolated from tumor-infiltrating lymphocytes (TILs) in acral melanoma, using cDNA expression cloning and identified a novel CT antigenic epitope encoded by MAGE-A6 with a single nucleotide polymorphism (SNP). This SNP conferred immunogenicity to the epitope, eliciting a robust immune response against tumor cells. While antigen identification has increasingly relied on reverse immunology approaches using reference sequences that do not contain SNPs, forward immunology approaches, such as cDNA expression cloning, directly identify antigens recognized by T cells exhibiting immune responses, enabling the detection of SNP-derived epitopes. Furthermore, in hot tumors such as acral melanoma that are characterized by a low tumor mutational burden, but high TIL infiltration, TILs predominantly respond to shared antigens with high immunogenicity. These findings underscore the utility of forward immunology in antigen discovery and highlight the potential of SNP-dependent tumor antigens in cancer immunotherapy.

  DOI PubMed

• The size of CD8+ infiltrating T cells is a prognostic marker for esophageal squamous cell carcinoma.

  Kengo Shigehara, Noriko Kawai, Tomohide Shirosaki, Yuma Ebihara, Aiko Murai, Akari Takaya, Serina Tokita, Kenta Sasaki, Naoki Shijubou, Terufumi Kubo, Kenji Murata, Takayuki Kanaseki, Tomohide Tsukahara, Yutaka Hatanaka, Tomoko Mitsuhashi, Toshiaki Shichinohe, Yoshihiko Hirohashi, Satoshi Hirano, Toshihiko Torigoe

  Scientific reports   15 ( 1 ) 26638 - 26638  2025.07  [International journal]

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  In many malignancies, an increased number of tumor-infiltrating lymphocytes (TILs) is recognized as a favorable prognostic factor, with exceptions such as renal cell carcinoma. However, the clinical significance of TIL size remains unclear. T-cell activation by mitogens increases cell size, partly via c-myc expression, suggesting that larger T cells may be more activated. We hypothesized that TIL size might be prognostically relevant in cancer patients. Here, we examined the relationship between the size and number of tumor-infiltrating CD8 + T cells and patient prognosis in 96 cases of esophageal squamous cell carcinoma (ESCC). We employed artificial intelligence (AI) analysis to quantify the mean size of intratumoral CD8+ T cells in each sample. Patients were then divided into "Large" and "Small" CD8+ T cell groups according to the median T-cell size. Similarly, we classified cases into "High" and "Low" groups based on CD8 + T-cell numbers. We found that patients in the Large CD8+ T cell group had significantly better overall survival than those in the Small CD8+ T cell group by a univariate analysis (p = 0.039), but the difference did not reach statistical significance in a multivariate analysis (p = 0.054). Patients in the High CD8 + T cell group had better outcomes than those in the Low CD8+ T cell group. There was no significant correlation between CD8+ T cell size and count, and their combination (Large/High) identified a subgroup of patients with the most favorable prognosis. Our findings suggest that CD8+ T cell size could serve as an independent prognostic marker in ESCC.

  DOI PubMed

• SARS-CoV-2 infection promotes lung thrombosis by inducing integrinβ3 expression in vascular endothelial cells.

  Wataru Ito, Yuya Sakurai, Nako Maishi, Ryo Takeda, Takahito Teshirogi, Li Yu, Yasuhiro Hida, Michihito Sasaki, Yasuko Orba, Takuya Tsumita, Haruhisa Watanabe, Tadahiro Iimura, Terufumi Kubo, Shinsuke Toba, Akihiko Sato, Aya Matsuda, Daisuke Kyuno, Makoto Osanai, Yoichi Ohiro, Toshihiko Torigoe, Hirofumi Sawa, Kyoko Hida

  Scientific reports   15 ( 1 ) 20447 - 20447  2025.07  [International journal]

  　View Summary

  Severe COVID-19 shows a high incidence of pulmonary thrombosis. However, the molecular mechanism underlying this phenomenon remains unclear. We have performed RNA sequencing of isolated endothelial cells (ECs) from infected mid-aged and young mice. Compared to young mice, Integrinβ3 (ITGB3) expression levels were higher in ECs of mid-aged mice which showed thrombosis in lungs. SARS-CoV-2 exposure increased the number of adhered platelets on the EC monolayer in vitro. Knockdown of ITGB3 in ECs decreased platelet adhesion to them. Among the molecules known as SARS-CoV-2 receptors, Kringle-containing transmembrane protein 1 contributed to ITGB3 upregulation in ECs by SARS-CoV-2. Histological analysis showed that ITGB3-positive blood vessels were frequently detected not only in infected-mid-aged mouse lungs but also in COVID-19-affected human autopsy lungs. This study suggests that the induction of ITGB3 expression in ECs is one of the mechanisms of thrombosis in severe COVID-19 pneumonia.

  DOI PubMed

• Large number of CD68+/CD163+ Hofbauer cells and characteristic vascular structural alterations in the placental villi of cases with placenta accreta spectrum.

  Hazuki Kashiwagi, Kengo Shigehara, Terufumi Kubo, Yoshihiko Hirohashi, Tasuku Mariya, Kazuhiko Matsuo, Tomoyuki Minowa, Shin-Ichi Ishioka, Kenji Murata, Takayuki Kanaseki, Tomohide Tsukahara, Tadashi Hasegawa, Tsuyoshi Saito, Toshihiko Torigoe

  Medical molecular morphology    2025.03  [Domestic journal]

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  Placenta accreta spectrum (PAS) is a serious disease leading to complications and maternal death. The objective of the study was to characterize the placental villi and blood vessels of PAS villi histopathologically. We investigated 10 cases of PAS (five cases of placenta increta, two cases of placenta accreta, and three cases of placenta percreta) histologically. Immunohistochemical staining using anti-CD68 or anti-CD163 antibodies was performed to detect and count Hofbauer cells. Immunohistochemical staining with an anti-CD34 antibody was used to detect vascular endothelial cells, and the number and area of vessels were analyzed. The numbers of CD68-positive or CD163-positive Hofbauer cells were larger in PAS cases compared with control cases. The vascular area in villi was smaller in PAS cases compared with control cases. The number of blood vessels in villi was slightly higher in PAS cases than in control cases. The numbers of Hofbauer cells and vessels in villi were larger in PAS cases compared with control cases, whereas the area of vessels in villi was smaller in PAS cases compared with control cases. Although their biological meaning is elusive, these findings provide novel insights into the pathogenesis of PAS, particularly regarding the role of Hofbauer cells in immune-suppressive role and angiogenesis and the alterations in vascular structure and hemodynamics in the chorionic villi.

  DOI PubMed

• Development of a T cell engaging bispecific antibody targeting long non-coding RNA PVT1.

  Hirotaka Kato, Tomohide Tsukahara, Kenji Murata, Hiromu Nishikata, Yuka Mizue, Takashi Sasaya, Terufumi Kubo, Takayuki Kanaseki, Yoshihiko Hirohashi, Atsushi Oyagi, Tatsuo Maeda, Akihiro Miyazaki, Toshihiko Torigoe

  Cancer immunology, immunotherapy : CII   74 ( 4 ) 133 - 133  2025.03  [International journal]

  　View Summary

  The development of effective immunotherapies for solid tumors remains a significant challenge. In previous studies, we identified PVT1, a long non-coding RNA, with the peptide HF10 derived from PVT1, presented by HLA-A24. This study aims to develop a single-chain variable fragment (scFv) that specifically recognizes the HLA-A24/HF10 complex (HF10 scFv) and to evaluate its specificity, reactivity, and therapeutic potential as part of a T cell engaging bispecific antibody (HF10xCD3) in vitro and in vivo. Using a scFv phage display library, we screened for scFv clones targeting the HLA-A24/HF10 peptide complex. The selected HF10 scFv was engineered into an IgG1 format (HF10-hIgG1), which demonstrated high affinity (KD = 2.18 × 10⁻⁸ M) and specific detection of the HLA-A24/HF10 complex on HLA-A24( +)/PVT1( +) tumor cell lines. Furthermore, HF10 scFv was incorporated into a T cell engaging bispecific antibody (HF10xCD3), which induced cytotoxicity in these tumor cell lines. In a mouse xenograft model, HF10xCD3 administration exhibited significant anti-tumor activity. In conclusion, HF10xCD3 represents a promising candidate for immunotherapy targeting solid tumors.

  DOI PubMed

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Research Projects 【 display / non-display 】

Research Projects 【 display / non-display 】

• Deciphering the mechanism of type 2 inflammation environment formation initiated by p63/p73 expressed in airway epithelial cells

  Grant-in-Aid for Scientific Research (C)

  Project Year :

  2024.04

  -

  2027.03

   

  久保 輝文

• Comprehensive analysis of T-cell receptor ligand by using pHLA multimer library

  Grant-in-Aid for Challenging Research (Pioneering)

  Project Year :

  2021.07

  -

  2025.03

   

  鳥越 俊彦, 久保 輝文

  　View Summary

  (1)Q-dot pHLA multimerの合成と性能評価 本年度は、Q-dot標識pHLA multimerの合成を実施し、性能評価を行った。ペプチド・HLA複合体のアセンブリには合成ペプチドチトとEmpty HLA multimerを混合する方法によって実施した。Q-dot標識pHLA multimerが、蛍光標識pHLA multimerを上回る感度を持つことを確認した。本技術については特許出願を準備中。barcode DNAを結合したpHLA multimerライブラリーを合成し、末梢血T細胞プールと反応させた。pHLA multimer陽性T細胞をソーティングし、barcode DNAのシークエンス解析によってT細胞に結合したpHLA multimerの種類と結合数を分析した。 (2)ヒト腫瘍浸潤T細胞を用いたTCR解析 昨年度に引き続き、今年度はヒト大腸がん組織の腫瘍浸潤T細胞を対象に、pHLA multimerライブラリー(蛍光標識)と反応させ、 pHLA multimerの結合したT細胞をフローサイトメーターによってソーティングした。がん精巣抗原に特異的なTCRをもつT細胞の検出を確認した。さらに、抗原特異的T細胞のシングルセルTCR遺伝子解析を実施し、TCRa鎖、b鎖のレパトワー分析を実施した。 (3)ヒト末梢血T細胞のTCRレパトワー解析とpHLA multimer解析 非小細胞肺がん患者の末梢血T細胞を分離し、TCR遺伝子解析によるレパトワー解析を実施した。化学放射線療法の前後でTCRレパトワーが変化した患者群では、その後に行われた抗PD-L1抗体治療の効果が高いことを見出した。新型コロナウイルス抗原pHLA multimerを作成し、健常人末梢血T細胞からウイルス特異的T細胞の検出に成功した。特許出願を実施。

• Comprehensive understanding of p63/p73-dependent deviated state keratinocytes in atopic dermatitis.

  Grant-in-Aid for Early-Career Scientists

  Project Year :

  2019.04

  -

  2022.03

   

  KUBO Terufumi

  　View Summary

  In this study, we aimed to investigate the influence of type 2 inflammatory environment on ΔNp63 expression and AD associated molecules regulated by ΔNp63 in keratinocyte. In healthy skin tissue, we observed inversed expression pattern between ΔNp63 and some barrier related proteins including filaggrin, caspase-14, claudin-1 and claudin-4. ΔNp63 regulated expression of these genes and proteins. In addition, production of IL-1β and IL-33, pro-inflammatory cytokines, were also modulated by ΔNp63. IL-13 interfered ΔNp63 down-regulation during calcium induced keratinocyte differentiation. IL-13 modulated some barrier and inflammation related molecules, which were under regulation of ΔNp63. We propose the IL-13-ΔNp63 axis would integrate two major factors of AD pathogenesis, epidermal barrier dysregulation and increased cytokine production of keratinocyte.

• Basic research on the immunohistological categorization of tumor microenvironment

  Grant-in-Aid for Scientific Research (A)

  Project Year :

  2017.04

  -

  2020.03

   

  TORIGOE Toshihiko

  　View Summary

  Tumor microenvironment (TME) was categorized into three classes by using tissue single cell analysis and multiplex immunohistochemistry (IHC) of primary human cancer tissues. It was revealed that seven IHC biomarkers were suitable for the TME classification, which could become the predictive marker for immune checkpoint blockade therapy as well as patients’ prognosis. In addition, clonality of T-cell antigen receptors and their target antigens were analyzed in primary cancer tissues. It was found that there was a close correlation between TME classification and T-cell clonality. We proposed a novel diagnostic tool that should contribute to the precision therapy of cancer.

• Mechanism of the ILC2 activation in bronchial asthma regulated by deltaNp63-related epimmunome

  Grant-in-Aid for Young Scientists (B)

  Project Year :

  2016.04

  -

  2019.03

   

  KUBO Terufumi, TSUJIWAKI Mitsuhiro, HIROHASHI Yoshihiko, TSUKAHARA Tomohide, KANASEKI Takayuki, NAKATSUGAWA Munehide, HASEGAWA Tadashi, TORIGOE Toshihiko

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  ΔNp63-positive basal epithelial cells were covered with differentiated ΔNp63-negative cells in healthy bronchial tissue; however, ΔNp63-positive cells were directly exposed to the bronchial lumen due to severe epithelial shedding in the asthmatic airway. ΔNp63 regulated bronchial apoptosis in response to Toll-like receptor 3 stimulation. Expression of ΔNp63 was modulated by stimulation with the certain type of protease signal. On the other hand, ΔNp63 controlled the transcriptional expression and protein release of some epithelium-derived proinflammatory cytokines and endogenous protease inhibitors.

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