Research activity information

• Mar. 2017 日本薬理学会, 年会優秀発表賞

  長坂 明臣

• Alteration of long and short-term hematopoietic stem cell ratio causes myeloid-biased hematopoiesis

  Katsuyuki Nishi, Taro Sakamaki, Akiomi Nagasaka, Kevin S Kao, Kay Sadaoka, Masahide Asano, Nobuyuki Yamamoto, Akifumi Takaori-Kondo, Masanori Miyanishi

  Jun. 2025


• Isolation Method for Long-Term and Short-Term Hematopoietic Stem Cells.

  Katsuyuki Nishi, Akiomi Nagasaka, Taro Sakamaki, Kay Sadaoka, Masanori Miyanishi

  Self-renewal capacity and multi-lineage differentiation potential are generally regarded as the defining characteristics of hematopoietic stem cells (HSCs). However, numerous studies have suggested that functional heterogeneity exists in the HSC compartment. Recent single-cell analyses have reported HSC clones with different cell fates within the HSC compartment, which are referred to as biased HSC clones. The mechanisms underlying heterogeneous or poorly reproducible results are little understood, especially regarding the length of self-renewal when purified HSC fractions are transplanted by conventional immunostaining. Therefore, establishing a reproducible isolation method for long-term HSCs (LT-HSCs) and short-term HSCs (ST-HSCs), defined by the length of their self-renewal, is crucial for overcoming this issue. Using unbiased multi-step screening, we identified a transcription factor, Hoxb5, which may be an exclusive marker of LT-HSCs in the mouse hematopoietic system. Based on this finding, we established a Hoxb5 reporter mouse line and successfully isolated LT-HSCs and ST-HSCs. Here we describe a detailed protocol for the isolation of LT-HSCs and ST-HSCs using the Hoxb5 reporter system. This isolation method will help researchers better understand the mechanisms of self-renewal and the biological basis for such heterogeneity in the HSC compartment.

  May 2023, Journal of visualized experiments : JoVE, (195) (195), English, International magazine

  [Refereed]

  Scientific journal


• VGLL3 is a mechanosensitive protein that promotes cardiac fibrosis through liquid–liquid phase separation

  Yuma Horii, Shoichi Matsuda, Chikashi Toyota, Takumi Morinaga, Takeo Nakaya, Soken Tsuchiya, Masaki Ohmuraya, Takanori Hironaka, Ryo Yoshiki, Kotaro Kasai, Yuto Yamauchi, Noburo Takizawa, Akiomi Nagasaka, Akira Tanaka, Hidetaka Kosako, Michio Nakaya

  Abstract Myofibroblasts cause tissue fibrosis by producing extracellular matrix proteins, such as collagens. Humoral factors like TGF-β, and matrix stiffness are important for collagen production by myofibroblasts. However, the molecular mechanisms regulating their ability to produce collagen remain poorly characterised. Here, we show that vestigial-like family member 3 (VGLL3) is specifically expressed in myofibroblasts from mouse and human fibrotic hearts and promotes collagen production. Further, substrate stiffness triggers VGLL3 translocation into the nucleus through the integrin β1-Rho-actin pathway. In the nucleus, VGLL3 undergoes liquid-liquid phase separation via its low-complexity domain and is incorporated into non-paraspeckle NONO condensates containing EWS RNA-binding protein 1 (EWSR1). VGLL3 binds EWSR1 and suppresses miR-29b, which targets collagen mRNA. Consistently, cardiac fibrosis after myocardial infarction is significantly attenuated in Vgll3-deficient mice, with increased miR-29b expression. Overall, our results reveal an unrecognised VGLL3-mediated pathway that controls myofibroblasts’ collagen production, representing a novel therapeutic target for tissue fibrosis.

  Springer Science and Business Media LLC, Feb. 2023, Nature Communications, 14(1) (1)

  [Refereed]

  Scientific journal


• GRK5 ‐mediated inflammation and fibrosis exert cardioprotective effects during the acute phase of myocardial infarction

  Akiomi Nagasaka, Tsuyoshi Terawaki, Makoto Noda, Miyuki Takashima, Mika Fujino, Yuto Yamauchi, Shigeki Arawaka, Takeo Kato, Michio Nakaya

  Wiley, Jan. 2023, FEBS Open Bio, 13(2) (2), 380 - 391

  [Refereed]

  Scientific journal


• GPRC5B promotes collagen production in myofibroblasts

  Noburo Takizawa, Takanori Hironaka, Kyosuke Mae, Tomoyuki Ueno, Yuma Horii, Akiomi Nagasaka, Michio Nakaya

  Elsevier BV, Jul. 2021, Biochemical and Biophysical Research Communications, 561, 180 - 186

  [Refereed]

  Scientific journal


• Increased Expression of Heparan Sulfate 6-O-Sulfotransferase-2 Promotes Collagen Production in Cardiac Myofibroblasts

  Kotaro Kasai, Yuma Horii, Takanori Hironaka, Kyosuke Mae, Tomoyuki Ueno, Akiomi Nagasaka, Michio Nakaya

  Pharmaceutical Society of Japan, 2021, BPB Reports, 4(3) (3), 85 - 91

  [Refereed]

  Scientific journal


• Drebrin is induced during myofibroblast differentiation and enhances the production of fibrosis-related genes.

  Takanori Hironaka, Tomoyuki Ueno, Kyosuke Mae, Chikashi Yoshimura, Takumi Morinaga, Yuma Horii, Akiomi Nagasaka, Hitoshi Kurose, Michio Nakaya

  Fibrosis is attributed to excess deposition of extracellular matrix (ECM) proteins including collagen and is associated with various organ dysfunction. This excessive ECM is produced by myofibroblasts, which are differentiated from various cells by a variety of stimuli, represented by TGF-β. However, molecular mechanisms for the regulation of ECM production in myofibroblasts remain obscure. In this study, we demonstrate that the expression of drebrin, which binds to and increases the stability of actin filament in neurons, is increased in mouse hearts and lungs upon fibrosis. Drebrin is mainly expressed in myofibroblasts in the fibrotic hearts and lungs and promotes the expression of fibrosis-related genes, such as Acta2 and Col1a1. Taken together, our study identifies drebrin as a molecule that promotes the production of fibrosis-related genes in myofibroblasts.

  Aug. 2020, Biochemical and biophysical research communications, 529(2) (2), 224 - 230, English, International magazine

  [Refereed]

  Scientific journal


• Efferocytosis during myocardial infarction.

  Chikashi Yoshimura, Akiomi Nagasaka, Hitoshi Kurose, Michio Nakaya

  Myocardial infarction is one of the major causes of death worldwide. Many heart cells die during myocardial infarction through various processes such as necrosis, apoptosis, necroptosis, autophagy-related cell death, pyroptosis and ferroptosis. These dead cells in infarcted hearts expose the so-called 'eat-me' signals, such as phosphatidylserine, on their surfaces, enhancing their removal by professional and non-professional phagocytes. Clearance of dead cells by phagocytes in the diseased hearts plays a crucial role in the pathology of myocardial infarction by inhibiting the inflammatory responses caused by the leakage of contents from dead cells. This review focuses on the rapidly growing understanding of the molecular mechanisms of dead cell phagocytosis, termed efferocytosis, during myocardial infarction, which contributes to the pathophysiology of myocardial infarction.

  Jul. 2020, Journal of biochemistry, 168(1) (1), 1 - 6, English, International magazine

  [Refereed]

  Scientific journal


• Leukotriene B4 receptor 1 exacerbates inflammation following myocardial infarction.

  Yuma Horii, Michio Nakaya, Hiroki Ohara, Hiroaki Nishihara, Kenji Watari, Akiomi Nagasaka, Takeo Nakaya, Yuki Sugiura, Toshiaki Okuno, Tomoaki Koga, Akira Tanaka, Takehiko Yokomizo, Hitoshi Kurose

  Leukotriene B4 receptor 1 (BLT1), a high-affinity G-protein-coupled receptor for leukotriene B4 (LTB4 ), is expressed on various inflammatory cells and plays critical roles in several inflammatory diseases. In myocardial infarction (MI), various inflammatory cells are known to be recruited to the infarcted area, but the function of BLT1 in MI is poorly understood. Here, we investigated the role of BLT1 in MI and the therapeutic effect of a BLT1 antagonist, ONO-4057, on MI. Mice with infarcted hearts showed increased BLT1 expression and LTB4 levels. BLT1-knockout mice with infarcted hearts exhibited attenuated leukocyte infiltration, proinflammatory cytokine production, and cell death, which led to reduced mortality and improved cardiac function after MI. Bone-marrow transplantation studies showed that BLT1 expressed on bone marrow-derived cells was responsible for the exacerbation of inflammation in infarcted hearts. Furthermore, ONO-4057 administration attenuated the inflammatory responses in hearts surgically treated for MI, which resulted in reduced mortality and improved cardiac function after MI. Our study demonstrated that BLT1 contributes to excessive inflammation after MI and could represent a new therapeutic target for MI.

  May 2020, FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(6) (6), 8749 - 8763, English, International magazine

  [Refereed]

  Scientific journal


• An Assay to Determine Phagocytosis of Apoptotic Cells by Cardiac Macrophages and Cardiac Myofibroblasts

  Horii Y, Matsuda S, Watari K, Nagasaka A, Kurose H, Nakaya M

  Sep. 2017, bio-protocol, 7(18) (18)

  [Refereed]


• Phagocytosis Assay of Necroptotic Cells by Cardiac Myofibroblasts

  Horii Y, Matsuda S, Watari K, Nagasaka A, Kurose H, Nakaya M

  Sep. 2017, bio-protocol, 7(18) (18), English

  [Refereed]


• The proton-sensing G protein-coupled receptor T-cell deathassociated gene 8 (TDAG8) shows cardioprotective effects against myocardial infarction

  Akiomi Nagasaka, Chihiro Mogi, Hiroki Ono, Toshihide Nishi, Yuma Horii, Yuki Ohba, Koichi Sato, Michio Nakaya, Fumikazu Okajima, Hitoshi Kurose

  Aug. 2017, SCIENTIFIC REPORTS, 7(1) (1), 7812, English

  [Refereed]

  Scientific journal


• Cardiac myofibroblast engulfment of dead cells facilitates recovery after myocardial infarction

  Michio Nakaya, Kenji Watari, Mitsuru Tajima, Takeo Nakaya, Shoichi Matsuda, Hiroki Ohara, Hiroaki Nishihara, Hiroshi Yamaguchi, Akiko Hashimoto, Mitsuho Nishida, Akiomi Nagasaka, Yuma Horii, Hiroki Ono, Gentaro Iribe, Ryuji Inoue, Makoto Tsuda, Kazuhide Inoue, Akira Tanaka, Masahiko Kuroda, Shigekazu Nagata, Hitoshi Kurose

  Jan. 2017, JOURNAL OF CLINICAL INVESTIGATION, 127(1) (1), 383 - 401, English

  [Refereed]

  Scientific journal


• GRK6 phosphorylates I kappa B alpha at Ser(32)/Ser(36) and enhances TNF-alpha-induced inflammation

  Yuki Ohba, Michio Nakaya, Kenji Watari, Akiomi Nagasaka, Hitoshi Kurose

  May 2015, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 461(2) (2), 307 - 313, English

  [Refereed]

  Scientific journal


• Novel functions of GRK6 in macrophages by phosphorylating the non-GPCRs substrates

  Akiomi Nagasaka, Yuki Ohba, Hitoshi Kurose, Michio Nakaya

  2015, Macrophages, (2) (2)

  [Refereed][Invited]


• Apaf-1-independent programmed cell death in mouse development

  A. Nagasaka, K. Kawane, H. Yoshida, S. Nagata

  Jun. 2010, CELL DEATH AND DIFFERENTIATION, 17(6) (6), 931 - 941, English

  [Refereed]

  Scientific journal


• Degradation of nuclear DNA by DNase II-like acid DNase in cortical fiber cells of mouse eye lens

  Masaki Nakahara, Akiomi Nagasaka, Masato Koike, Kaori Uchida, Kohki Kawane, Yasuo Uchiyama, Shigekazu Nagata

  Jul. 2007, FEBS JOURNAL, 274(12) (12), 3055 - 3064, English

  [Refereed]

  Scientific journal

• 免疫不全マウスモデルにおけるLineage特異的なin vivo細胞分化法に関する基礎的検討

  長坂明臣, 酒巻太郎, 宮西正憲

  2025, 日本再生医療学会総会(Web), 24th


• 多層性解析によるヒト造血幹細胞単離法の最適化

  宮西正憲, 宮西正憲, 長坂明臣, 酒巻太郎, 西克幸, 塚本成幸, 定岡恵, 山本暢之, 井上翔太郎, MY An Le

  2024, 日本再生医療学会総会(Web), 23rd


• マクロファージにおけるpH感知性GPCRの生理的役割

  長坂 明臣, 黒瀬 等

  Oct. 2018, アグリバイオ, 2(11) (11), 79 - 83


• マクロファージにおけるpH低下の生理的意義

  長坂 明臣, 黒瀬 等

  (株)ニュー・サイエンス社, Jul. 2018, Medical Science Digest, 44(8) (8), 405 - 409, Japanese

  Introduction scientific journal

• 免疫不全マウスモデルにおけるLineage特異的なin vivo細胞分化法に関する基礎的検討

  長坂 明臣, 酒巻 太郎, 宮西正憲

  第24回日本再生医療学会総会, Mar. 2025


• The roles of T-cell death-associated gene 8 (TDAG8), a proton-sensing GPCR, in myocardial infarction

  長坂明臣

  Translational Research on Metabolic and Inflammatory Diseases and Cancer Date, Dec. 2019, English

  Oral presentation


• プロトン感知性受容体(TDAG8)に着目した心筋梗塞時におけるpH低下の生理的影響の解析

  長坂 明臣, 小野達貴, 仲矢 道雄, 黒瀬 等

  次世代を担う創薬・医療薬理シンポジウム2017, Aug. 2017, Japanese


• 心筋梗塞時におけるプロトン感知性受容体TDAG8の役割解析

  長坂 明臣, 小野 達貴, 仲矢 道雄, 黒瀬 等

  第90回 日本薬理学会年会, Mar. 2017


• DNase II欠損マウスを用いたマウスの発生段階におけるアポトーシス細胞の検出

  長坂 明臣, 川根 公樹, 長田 重一

  日本分子生物学会, Dec. 2008, Japanese

• THE MOLECULAR BIOLOGY SOCIETY OF JAPAN


• THE JAPANESE PHARMACOLOGICAL SOCIETY


• THE PHARMACEUTICAL SOCIETY OF JAPAN


• 日本組織培養学会


• 日本再生医療学会

• 血管内皮細胞障害を改善する新規機能分子の取得

  長坂 明臣

  日本学術振興会, 科学研究費助成事業, 基盤研究(C), 国立研究開発法人理化学研究所, 01 Apr. 2022 - 31 Mar. 2025


• 心肥大の退縮を担う分子の探索とその分子メカニズムの解明

  長坂 明臣

  文部科学省, 科学研究費補助金（基盤研究（C））, Apr. 2019 - Mar. 2022, Principal investigator

  Competitive research funding


• Analysis of the roles and interaction of various cells including myofibroblasts involved in fibrosis after myocardial infarction

  KUROSE HITOSHI

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (A), Kyushu University, 01 Apr. 2017 - 31 Mar. 2021

  Lesion of the heart after myocardial infarction is compensated by collagen. However, excess deposition of collagen is called as fibrosis that is a target of the treatment. Collagen is produced by myofibroblasts. Myocardial infarction results in death of cells at ischemic area, which causes inflammation. Inflammation is induced in parallel to progression of fibrosis that is controlled by myofibroblasts. Then, mechanistic analysis of inflammation is thought to be important to treat fibrosis. In addition to inflammation, concentration of proton is known to increase under the ischemic conditions. However, the physiological meaning has not been analyzed. In this project, importance of infiltration of leukocytes to ischemic area, the roles of proton-sensing receptor in myocardial infarction and receptor-mediated intracellular signaling were studied. Then, the association of these responses with fibrosis was evaluated.


• 腫瘍関連マクロファージにおけるプロトン感知性受容体の生理的役割解析

  長坂 明臣

  Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research（若手研究（B））, Apr. 2017 - Mar. 2018, Principal investigator

  Competitive research funding


• Roles of extracellular environment and various cells carrying different function for cardiac fibrosis

  KUROSE HITOSHI

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (A), Kyushu University, 21 Oct. 2013 - 31 Mar. 2017

  Heart often develops heart failure after myocardial infarction, if proper revascularization is not provided. Inflammatory responses are believed to be a factor to progress to heart failure after myocardial infarction. As inflammation is induced by the cellular components released from dead cells, the quick and efficient removal of dead cells in myocardial infarction is important for preventing the development of heart failure. So far, phagocytes such as macrophages and dendritic cells infiltrating infarct area are supposed to be the players of removing dead cells. In this study, myofibroblasts remove dead cell as well as phagocytes do. The removal of dead cells by myofibroblasts requires MFG-E8 that bridges between apoptotic cells and phagocytes. Thus, myofibroblasts remove dead cells after myocardial infarction, and suppress the progression to heart failure.


• 心筋梗塞におけるプロトン感知性受容体「GPR4」の生理的役割の解析

  長坂 明臣

  Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research（若手研究（B））, Apr. 2014 - Mar. 2016, Principal investigator

  Competitive research funding