ヒト心臓内幹細胞の増殖・分化誘導因子の発見と心筋分化初期プライミング因子の探索

2008 Fiscal Year Annual Research Report

• Project/Area Number: 20249046
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: 松原 弘明 Kyoto Prefectural University of Medicine, 医学研究科, 教授 (10239072)
• Co-Investigator(Kenkyū-buntansha): 高橋 知三郎 京都府立医科大学, 医学研究科, 助教 (20381973) ; 的場 聖明 京都府立医科大学, 医学研究科, 助教 (10305576) ; 天野 克也 京都府立医科大学, 医学研究科, 研究員 (70468263)
• Keywords: 再生医療 / 胚性幹細胞 / 心筋再生

Research Abstract

私達はヒト心筋生検組織より心筋分化可能な多能性間葉系幹細胞(CSC)の単離・増幅に成功した。この幹細胞の心筋分化効率は1-2%と低く、増殖能も遅く、世界中が心筋分化誘導・増殖促進因子を探索している。申請者らが心筋より単離成功した新規の心筋分化・増殖誘導因子(Cv2, MuRC)を中心に、初期心筋分化プライミング因子群の解析を実施した。
(Cv2) cDNAサブトラクションスクリーニング法とディファレンシャルスクリーニング法を組み合わせる事によって、「初期プライミング因子」群の候補になりえる6因子同定した。そのなかでも、比較的新規とされる骨形成タンパク質(BMP)結合タンパク質であるcrossveinless-2(Cv2)に注目し解析を進めている。
(MuRC)心臓Z-disk特異的に発現するmuscle-restricted coiled-coil rotein (MuRC)の解析による心筋分化誘導メカニズム解明心筋細胞のサルコメアの両端に位置するZ-discは、心筋の伸展を感知し、肥大や生存のためのシグナルを伝達し、心不全発症に重要な役割を担っている。申請者らは、成人心臓に発現している遺伝子の網羅的な探索を行い、心臓、骨格筋、血管平滑筋にのみ発現している筋細胞特異的新規遺伝子MuRCの単離に成功した。MuRCはactininの局在と一致して心筋Z-discに存在した。Tg-MuRCは著明な心筋細胞肥大と心電図上PR間隔の延長を認め、完全房室ブロックや心房細動を呈した。さらに、MuRC-Tgは心筋間質の著明な線維化と拡張した左心室径および低下した心駆出率も示した。MuRcはZ-disk存在蛋白であるため、まずMuRc-Tgで発症した心筋肥大誘導と心房細動の原因遺伝子の同定や心不全という表現型解析のため、MuRC-Tg群とNT-g群の左房と左室のそれぞれからmRNAを回収し、マイクロアレイにて遺伝子群を網羅的に検索する。MuRCはZ-Diskに存在するので、yeast-two hybrid法にて結合蛋白も同定する。

Research Products

• [Journal Article] Controlled Delivery of Basic Fibroblast Growth Factor Promotes Human Cardiosphere-Derived Cell Engraftment to Enhance Cardiac Repair forChronic Myocardial Infarction. (2008)
  • Author(s): Takehara N, Matsubara H., (他12人,最終著者)
  • Journal Title: J Am Coll Cardiol. 52 Pages: 1858-1865
  • Peer Reviewed
• [Journal Article] Downregulation of Dicer expression by serum withdrawal sensitizes humanendothelial cells to apoptosis. (2008)
  • Author(s): Asada S, Matsubara H., (他7人,8番目著者)
  • Journal Title: Am J Physiol Heart Circ Physiol. 295 Pages: 2512-2521
  • Peer Reviewed
• [Journal Article] Cardioprotective effects of erythropoietin in rats subjected toischemiareperfusion injury : assessment of infarct size with 99m Tc-anmexin V. (2008)
  • Author(s): Doue T, Matsubara H., (他6人,最終著者)
  • Journal Title: J Nucl Med. 49 Pages: 1694-1670
  • Peer Reviewed
• [Journal Article] Stemming heart failure with cardiac- or reprogrammed-stem cells. (2008)
  • Author(s): Tateishi K, Takehara N, Matsubara H, 0h H.
  • Journal Title: J Cell Mol Med 12 Pages: 2217-2232
  • Peer Reviewed
• [Journal Article] MURC, a muscle-restricted coiled-coil protein, is involved in the regulationof skeletal myogenesis. (2008)
  • Author(s): Tagawa M, Matsubara H., (他8人,9番目著者)
  • Journal Title: Am J PhysiolCell Physiol. 295 Pages: C490-C498
  • Peer Reviewed
• [Journal Article] Exercise-Induced Changes of Functional Mitral Regurgitation in Asymptomatic or Mildly Symptomatic Patients With Idiopathic Dilated Cardiomyopathy. (2008)
  • Author(s): Yamano T, Matsubara H., (他9人10番目著者)
  • Journal Title: Am J Cardiol 102 Pages: 481-485
  • Peer Reviewed
• [Journal Article] Crossveinless-2 controls bone morphogenetic protein signaling duringearly cardiomyocyte differentiation in P19 cells. (2008)
  • Author(s): Harada K, Ogai A, Takahashi T, Kitakaze M, Matsubara H, Oh H.
  • Journal Title: J Biol Chem. 283 Pages: 26705-26713
  • Peer Reviewed
• [Journal Article] MURC, a Muscle-Restricted Coiled-Coil Protein That Modulates the Rho/ROCK Pathway, Induces Cardiac Dysfunction and Conduction Disturbance. (2008)
  • Author(s): Ogata T, Matsubara H., (他8人,9番目著者)
  • Journal Title: Mol Cell Biol 28 Pages: 3424-3436
  • Peer Reviewed
• [Journal Article] Generation of reentrant arrhythmias by dominant-negative inhibition of connexin43 in rat cultured myocyte monlayers. (2008)
  • Author(s): Nakagami T, Matsubara H., (他7人8番目著者)
  • Journal Title: Cardiovasc Res. 79 Pages: 70-79
  • Peer Reviewed
• [Journal Article] Aldosterone nongenomically produces NADPH oxidase-dependent reactive oxygen species and induces myocyte apoptosis. (2008)
  • Author(s): Hayashi H, Matsubara H., (他8人,9番目著者)
  • Journal Title: Hypertens Res 31 Pages: 363-375
  • Peer Reviewed
• [Presentation] Calcium-Dependent Tyrosine Kinase PYK2 Plays Crucial Role in Angiotensin II - but Not Norepineprine-mediated Activation fo Rac/Superoxide Pathway that Differentially Regulates Blood Pressure and Vasoconstriction (2008)
  • Author(s): Matsui A, Okigaki M, Katsume A, Jisan C, Yamaguchi S, Yamada H, Matsubara H.
  • Organizer: American Heart Association (米国心臓病学術会議)
  • Place of Presentation: New Orleans, USA
  • Year and Date: 20081108-20081112
• [Presentation] Ca+/Redox-sensitive Tyrosine Kinase PYK2 Promotes Atherogenesis through OxLDL/ROS/p21-Dependent Premature Senescence and Cytokine Induction in Endothelium. (2008)
  • Author(s): Katsume A, Okigaki M, Matsui A, Yamaguchi S, Kishita E, Yamada H, Matsubara H
  • Organizer: American Heart Association (米国心臓病学術会議)
  • Place of Presentation: New Orleans, USA
  • Year and Date: 20081108-20081112
• [Presentation] Bone Morphogenetic Protein-2 plays Crucial Roles In The Formation of Medial Calcification And Atherosclerosis In Vivo. (2008)
  • Author(s): Nakagawa Y, Ikeda K, Nakano R, Uraoka M, Yutaka K, Koide M, Matsubara H
  • Organizer: American Heart Association (米国心臓病学術会議)
  • Place of Presentation: New Orleans, USA
  • Year and Date: 20081108-20081112
• [Presentation] Bone Marrow Angiotensin At1 Receptor Regulates Differentiation of Monocyte/Macrophage-Lineage Progenitors from Hematopoietic Stem Cells. (2008)
  • Author(s): Tsubakimoto Y, Yamada H, Yokoi H, Takata H, Kawahito H, Kishida S, Matsui A, Hirai H, Imanishi J, Ashi hara E, Maekawa T, Takah ashi T, Okigaki M, Matsubara H
  • Organizer: American Heart Association (米国心臓病学術会議)
  • Place of Presentation: New Orleans, USA
  • Year and Date: 20081108-20081112
• [Presentation] Bone Marrow ATI Receptor Augments Neointima Formation by Promoting Mobilization of Smooth Muscle Progenitor Cells in Platelet-Derived SDF-1α-Dependent Manner. (2008)
  • Author(s): Yokoi H, Yamada H, Tsuba kimoto Y, Takata H, Kawahito H, Kishida S, Matsui A, Hirai H, Imanishi J, Ashihara E, Maekawa T, Takahashi T, Okigaki M, Matsubara H.
  • Organizer: American Heart Association (米国心臓病学術会議)
  • Place of Presentation: New Orleans, USA
  • Year and Date: 20081108-20081112
• [Presentation] Molecular Identification and Characterization of a Novel Gene that Regulation Endothelial Cell Apoptosis and Angiogenesis. (2008)
  • Author(s): Ikeda K, Koide M, Uraoka M, Nakagawa Y, Nakano R, Matsubara H.
  • Organizer: American Heart Association (米国心臓病学術会議)
  • Place of Presentation: New Orleans, USA
  • Year and Date: 20081108-20081112
• [Presentation] Calcium/Redox Sensitive Tyrosine Kinase Pyk2 Aggravates Myocardial Ishemia by Promoting Inflammatory Reactions and Cell Apoptosis. (2008)
  • Author(s): Kishita E, Honsho S, Yamaguchi S, Katsume A, Matsui A, Matsunaga S, Okigaki M, Matsubara H.
  • Organizer: American Heart Association (米国心臓病学術会議)
  • Place of Presentation: New Orleans, USA
  • Year and Date: 20081108-20081112
• [Presentation] Therapeutic Angiogenesis by Mononuclear Cell Implantation for Critical Digit Ischemia of Patients with Connective Tissue Disea (2008)
  • Author(s): Takahashi M, Ishigatsubbo Y, Fujimoto K, Miyamoto M, Horie T, Aizawa Y, Murohara T, Matsubara H, Ikeda U.
  • Organizer: American Heart Association (米国心臓病学術会議)
  • Place of Presentation: New Orleans, USA
  • Year and Date: 20081108-20081112