Regulation of atheroscllerosis by lymphatic endothelial calpain sysmtems

• Project/Area Number: 19K08590
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53020:Cardiology-related
• Research Institution: Showa University
• Principal Investigator: Miyazaki Takuro 昭和大学, 医学部, 准教授 (80398693)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2020: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: カルパイン / リンパ管内皮細胞 / タンパク質分解 / 制御性T細胞 / 動脈硬化症 / 脂質異常症 / リンパ球／制御性T細胞 / TGF-beta / リンパ / 制御性T細胞 / リンパ

Outline of Research at the Start

申請者はこれまで血管機能を制御する極めてユニークな機構として、ストレス応答性細胞内プロテアーゼ「カルパイン」ファミリーを検証し、同機構が動脈硬化症、網膜症および腫瘍血管などの病態下で血管内皮細胞機能異常の主因となることを解明した。本研究提案では、リンパ管内皮細胞のタンパク質分解異常がいかにして動脈硬化症の形成に寄与するか、制御性T細胞との相互作用を中心に分子メカニズムの解明を目指す。

Outline of Final Research Achievements

This study investigated whether the calpain systems in lymphatic endothelial cells (LECs) associate with the LEC-lymphocyte interaction under hypercholesterolemia. Lipidomic analysis in hypercholesterolemic mice showed that several lysophospholipids, including lysophosphatidic acid (LPA), accumulated in the lymphatic environment. LPA enables the potentiation of calpain systems in cultured LECs, which limits their ability to stabilize regulatory T cells (Treg) without altering other T cell subsets. Targeting calpain systems in LECs expanded Tregs in the blood circulation under hypercholesterolemia, concomitant with the redistribution of Tregs in lymphoid organs. Moreover, LPA-induced calpain overactivation potentiated the IL-18/NF-kB/VCAM1 axis in LECs, thereby inhibiting lymphocyte mobility on the cells. Thus, calpain systems in LECs have a key role in controlling Treg stability and trafficking under hypercholesterolemia.

Academic Significance and Societal Importance of the Research Achievements

一連の結果から、リンパ管内皮細胞のカルパインシステムは、脂質異常症存在下のリンパ管の形態維持だけでなく、制御性T細胞の安定性や動態にも影響することが判明した。既報から、脂質異常症においてリンパ管がリポタンパク質や免疫細胞の輸送経路として重要であることは明らかとなっていたが、直接的な免疫細胞の制御に関わっていることは本研究において初めて解明された。本研究は、動脈硬化症の発症原因の包括的な理解に資するものと期待される。

Research Products

• [Journal Article] 血管系疾患に潜むカルパインシステムの制御異常 (2022)
  • Author(s): 宮崎拓郎, 宮崎章
  • Journal Title: 生化学 Volume: 94 Pages: 14-25
  • Peer Reviewed
• [Journal Article] Calpain-mediated proteolytic production of free amino acids in vascular endothelial cells augments obesity-induced hepatic steatosis. (2022)
  • Author(s): Risako Akasu, Takuro Miyazaki, Mohamed Z Elhussiny, Yuki Sugiura, Yuki Tomitsuka, Shogo Haraguchi, Kinya Otsu, Vishwajit S Chowdhury, Akira Miyazaki
  • Journal Title: The Journal of biological chemistry Volume: N/A Issue: 6 Pages: 101953-101953
  • DOI: 10.1016/j.jbc.2022.101953
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Hypercholesterolemia and Lymphatic Defects: The Chicken or the Egg? (2021)
  • Author(s): Takuro Miyazaki, Akira Miyazaki
  • Journal Title: Frontiers in Cardiovascular Medicine Volume: 8 Pages: 701229-701229
  • DOI: 10.3389/fcvm.2021.701229
  • Peer Reviewed / Open Access
• [Journal Article] Oral administration of L‐citrulline changes the concentrations of plasma hormones and biochemical profile in heat‐exposed broilers (2021)
  • Author(s): Chowdhury Vishwajit S.、Ouchi Yoshimitsu、Han Guofeng、Eltahan Hatem M.、Haraguchi Shogo、Miyazaki Takuro、Shiraishi Jun‐ichi、Sugino Toshihisa、Bungo Takashi
  • Journal Title: Animal Science Journal Volume: 92 Issue: 1
  • DOI: 10.1111/asj.13578
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Calpain proteolytic systems counteract endothelial cell adaptation to inflammatory environments. (2020)
  • Author(s): Miyazaki T, Akasu R, Miyazaki A.
  • Journal Title: Inflamm Regen. Volume: 40 Issue: 1 Pages: 5-5
  • DOI: 10.1186/s41232-020-00114-x
  • Peer Reviewed / Open Access
• [Journal Article] Hic-5 is required for activation of pancreatic stellate cells and development of pancreatic fibrosis in chronic pancreatitis (2020)
  • Author(s): Gao Lin、Lei Xiao-Feng、Miyauchi Aya、Noguchi Masahito、Omoto Tomokatsu、Haraguchi Shogo、Miyazaki Takuro、Miyazaki Akira、Kim-Kaneyama Joo-ri
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 19105-19105
  • DOI: 10.1038/s41598-020-76095-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Calpain-Associated Proteolytic Regulation of the Stromal Microenvironment in Cancer (2020)
  • Author(s): Takuro Miyazaki, Risako Akasu, Akira Miyazaki
  • Journal Title: Current Pharmaceutical Design Volume: N/A Issue: 28 Pages: 3128-3138
  • DOI: 10.2174/1381612827666210311143053
  • Peer Reviewed
• [Journal Article] Effects of lipid metabolism on mouse incisor dentinogenesis. (2020)
  • Author(s): Kurotaki, Y., Sakai, N., Miyazaki, T., Hosonuma, M., Sato, Y., Karakawa, A., Chatani, M., Myers, M., Suzawa, T., Negishi-Koga, T., Kamijo, R., Miyazaki, A., Maruoka, Y., Takami, M.
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 5102-5102
  • DOI: 10.1038/s41598-020-61978-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Endothelial calpain systems orchestrate myofibroblast differentiation during wound healing. (2019)
  • Author(s): Miyazaki T, Haraguchi S, Kim-Kaneyama JR, Miyazaki A.
  • Journal Title: FASEB Journal Volume: 33 Issue: 2 Pages: 2037-2046
  • DOI: 10.1096/fj.201800588rr
  • Peer Reviewed
• [Journal Article] L-Leucine increases the daily body temperature and affords thermotolerance in broiler chicks. (2019)
  • Author(s): Han G, Yang H, Wang Y, Haraguchi S, Miyazaki T, Bungo T, Tashiro K, Furuse M, Chowdhury VS
  • Journal Title: Asian-Australas J Anim Sci Volume: in press Issue: 6 Pages: 842-848
  • DOI: 10.5713/ajas.18.0677
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Thrombolytic Therapy for Acute Ischemic Stroke: Past and Future. (2019)
  • Author(s): Shibata K, Hashimoto T, Miyazaki T, Miyazaki A, Nobe K.
  • Journal Title: Curr Pharm Des Volume: in press Issue: 3 Pages: 242-250
  • DOI: 10.2174/1381612825666190319115018
  • Peer Reviewed
• [Journal Article] Evaluation of protein-protein interactions using an on-membrane digestion technique (2019)
  • Author(s): Obama Takashi、Miyazaki Takuro、Aiuchi Toshihiro、Miyazaki Akira、Itabe Hiroyuki
  • Journal Title: Journal of Visualized Experiments Volume: 149 Issue: 149 Pages: 149-149
  • DOI: 10.3791/59733
  • Peer Reviewed / Open Access
• [Journal Article] Light-at-night exposure affects brain development through pineal allopregnanolone-dependent mechanisms (2019)
  • Author(s): Haraguchi S, Kamata M, Tokita T, Tashiro KI, Sato M, Nozaki M, Okamoto-Katsuyama M, Shimizu I, Han G, Chowdhury VS, Lei XF, Miyazaki T, Kim-Kaneyama JR, Nakamachi T, Matsuda K, Ohtaki H, Tokumoto T, Tachibana T, Miyazaki A, Tsutsui K.
  • Journal Title: eLife Volume: 8
  • DOI: 10.7554/elife.45306
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Alleviation of murine osteoarthritis by deletion of the focal adhesion mechanosensitive adapter, Hic-5 (2019)
  • Author(s): Miyauchi Aya、Kim-Kaneyama Joo-ri、Lei Xiao-Feng、Chang Song Ho、Saito Taku、Haraguchi Shogo、Miyazaki Takuro、Miyazaki Akira
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 15770-15770
  • DOI: 10.1038/s41598-019-52301-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] リンパ管内皮細胞の細胞内プロテアーゼシステムによる動脈硬化病変の遠隔制御 (2021)
  • Author(s): 宮崎 拓郎
  • Organizer: 第21回抗加齢医学会総会
  • Invited
• [Presentation] Defective exon junction complex disturbs cholesterol handling in macrophages thereby exacerbating atherosclerosis (2021)
  • Author(s): Takuro Miyazaki
  • Organizer: 4th International Caparica Conference in SPLICING 2021
  • Int'l Joint Research / Invited
• [Presentation] Obesity-induced hepatic steatosis harbors proteolytic production of branched-chain amino acids in vascular endothelial cells (2021)
  • Author(s): Risako Akasu, Takuro Miyazaki, Akira Miyazaki
  • Organizer: 19th International Symposium on Atherosclerosis
  • Int'l Joint Research
• [Presentation] Hypercholesterolemic alterations of proteolytic systems in lymphatic endothelial cells abrogate immunosuppression in atheroprone arteries (2021)
  • Author(s): Takuro Miyazaki, Risako Akasu, Akira Miyazaki
  • Organizer: 19th International Symposium on Atherosclerosis
  • Int'l Joint Research
• [Presentation] カルパインプロテアーゼシステムによる動脈硬化性疾患の制御 (2021)
  • Author(s): 宮崎 拓郎
  • Organizer: 第94回日本生化学会大会
  • Invited
• [Presentation] Defective exon junction complex disturbs cholesterol handling in macrophages thereby exacerbating atherosclerosis (2020)
  • Author(s): Takuro Miyazaki
  • Organizer: Splicing2020
  • Int'l Joint Research / Invited
• [Presentation] カルパインによる血管内皮細胞機能異常が耐糖能におよぼす影響 (2020)
  • Author(s): 赤須 里沙子、宮崎 拓郎、冨塚 祐希、宮崎 章
  • Organizer: 第５２回日本動脈硬化学会学術集会
• [Presentation] しなやかな血管を手に入れる (2019)
  • Author(s): 宮崎 拓郎
  • Organizer: 平成31年健康大学第１回教養講座
  • Invited
• [Presentation] リンパ管のタンパク質恒常性低下を起点としたあらたな動脈硬化性血管炎の制御機構 (2019)
  • Author(s): 宮崎 拓郎
  • Organizer: 第40回日本炎症・再生医学会 シンポジウム２：炎症病としてのCardiovascular Diseases
  • Invited
• [Presentation] Emerging roles of calpain-6 in macrophage cholesterol handling and atherogenesis. FASEB SRC The biology of Calpains in Health and Disease. (2019)
  • Author(s): Miyazaki T
  • Organizer: FASEB SRC The biology of Calpains in Health and Disease. Session title「CALPAIN IN DIABETES AND ATHEROSCLEROSIS」
  • Int'l Joint Research / Invited
• [Presentation] リンパ管のタンパク質恒常性低下を起点としたあらたな動脈硬化性血管炎の制御機構 (2019)
  • Author(s): 宮崎 拓郎
  • Organizer: 第40回 日本炎症・再生医学会 シンポジウム２ 炎症病としてのCardiovascular diseases
  • Invited
• [Presentation] リンパ管内皮細胞を起点とした動脈硬化症の発症機構 (2019)
  • Author(s): 宮崎 拓郎、原口 省吾、森戸 大介、金山 朱里、宮崎 章
  • Organizer: 第51回 日本動脈硬化学会学術集会
• [Presentation] 脂質異常症に起因するリンパ管内皮細胞のタンパク質恒常性低下は動脈硬化症を増悪化する (2019)
  • Author(s): 宮崎拓郎、原口省吾、金山朱里、宮崎章
  • Organizer: 第61回 日本脂質生化学会大会
• [Presentation] 血管内皮カルパインが耐糖能異常におよぼす影響 (2019)
  • Author(s): 赤須里沙子、宮崎拓郎、冨塚祐希、宮崎章
  • Organizer: CVMW2019 心血管代謝週間
• [Presentation] 血管内皮カルパインが肥満誘発性耐糖能異常におよぼす影響 (2019)
  • Author(s): 冨塚祐希 宮崎拓郎 赤須里沙子 宮崎章
  • Organizer: 2019年度日本生化学会関東支部例会
• [Remarks] reserchmap_Takuro Miyazaki
  • URL: https://researchmap.jp/miyazakitakuro/?lang=japanese
• [Remarks] 宮崎拓郎（昭和大学医学部生化学講座）ホームページ
  • URL: http://www10.showa-u.ac.jp/~biochem/Takuro_Miyazaki/Takuro_Miyazaki.html
• [Remarks] Research map_Takuro Miyazaki
  • URL: https://researchmap.jp/miyazakitakuro/?lang=japanese
• [Remarks] 昭和大学医学部生化学講座ホームページ
  • URL: http://www10.showa-u.ac.jp/~biochem/Takuro_Miyazaki/Takuro_Miyazaki.html
• [Remarks] Reserchmap 宮崎拓郎
  • URL: https://researchmap.jp/miyazakitakuro/presentations
• [Remarks] Researchgate Takuro Miyazaki
  • URL: https://www.researchgate.net/profile/Takuro_Miyazaki2/