Investigation of the molecular mechanism of progenitor cell-mediated therapeutic angiogenesis

• Project/Area Number: 18390232
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Circulatory organs internal medicine
• Research Institution: Nagoya University
• Principal Investigator: MUROHARA Toyoaki Nagoya University, Graduate School of Medicine, Professor (90299503)
• Co-Investigator(Kenkyū-buntansha): KOMORI Kimihiro Nagoya University, Graduate School of Medicine, Professor (40225587) ; NUMAGUCHI Yasushi Nagoya University, School of Medicine, Associate Professor (90378224) ; SHINTANI Satoshi Nagoya University, University Hospital, Assistant Professor (20309777) ; 近藤 隆久 名古屋大学, 医学部附属病院, 講師 (00303644)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥17,450,000 (Direct Cost: ¥15,200,000、Indirect Cost: ¥2,250,000); Fiscal Year 2007: ¥9,750,000 (Direct Cost: ¥7,500,000、Indirect Cost: ¥2,250,000); Fiscal Year 2006: ¥7,700,000 (Direct Cost: ¥7,700,000)
• Keywords: regenerative medicine / angiogenesis / cell therapy / gene therapy / endothelial cell / stem cell / therapeutic angiogenesis / endothelial progenitor cell / 血管再生療法

Research Abstract

Therapeutic angiogenesis using antologous stem/progenitor cells represents a novel strategy for severely ischemic diseases. Recent reports indicated that adipose tissues could supply adipose-derived stem cells (ASCs). Accordingly, we examined whether implantation of ASCs would augment angiogenesis using a mouse model of hind limb ischemia. C57BL/6J mouse inguinal fat pad was obtained and ASCs were isolated using standard medium.
Fluorescence-activated cell sorter (FACS) analysis revealed that ASCs were Sca-1^+CD31-CD34-flk-1-c-kif-Lin. ASCs expressed vascular endothelial growth factor (VEGF) and stromal cell-derived factor I (SDF-1) mRNAs and proteins. Hind limb ischemia was surgically induced and culture-expanded ASCs(1.0xl0^6cells/animal), PBS or differentiated mature adipocytes(MAs) as control cells were injected into the ischemic tissues. At 3 weeks, the ASC group had a greater laser Doppler blood flow index (0.93±0.08 vs. 0.79±0.14 p<0.05) and a higher capillary density (1.51±0.08 vs. 1.13±0.07 p<0.01) compared to the control group. Implantation ofASCs increased circulating EPCs measured by FACS and culture assay. SDF-I mRNA abundance at ischemic tissues and serum SDF-1 levels were greater in the ASC group than control group. Finally, intraperitoneal injection of an anti-SDF-1 neutralizing antibody reduced therapeutic efficacies of ASCs implantation.
Adipose tissue would be a valuable source for cell-based therapeutic angiogenesis. Moreover, SDF-1 may play a pivotal role in the ASCs-mediated angiogenesis at least in part by facilitating mobilization of EPCs.

Research Products

• [Journal Article] Regulation of VEGF-mediated angiogenesis by the Akt/PKB substrate Girdin. (2008)
  • Author(s): Kitamura T, Asai N, Enomoto A, Maeda K, Kato T, Ishida M, Jiang P, Watanabe T, Usuknra J, Kondo T, Costantini F, Murohara T, Takahashi M.
  • Journal Title: Nat. Cell Biol. (in press)(印刷中)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Regulation of VEGF-mediated angiogenesis by the Akt/PKB substrate Girdin (2008)
  • Author(s): Kitamura, T, Asai, N, Enomoto, A, Maeda, K, Kato, T, Ishida, M, Jiang, P, Watanabe, T, Usukura, J, Kondo, T, Costantini, F, Murohara, T, Takahashi, M
  • Journal Title: Nat. Cell Biol (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Regulation of VEGF-mediated angiogenesis by the Akt/PKB substrate Girdin. (2008)
  • Author(s): Kitamura T, Asai N, Enomoto A, Maeda K, Kato T, Ishida M, Jiang P, Watanabe T, Usukura J, Kondo T, Costantini F, Murohara T, akahashi M
  • Journal Title: Nat. Cell Biol. (in press)
  • Peer Reviewed
• [Journal Article] Therapeutic angiogenesis using novel vascular endothelial growth factor-E/human placental growth factor chimera genes. (2007)
  • Author(s): Inoue N, Kondo T, Kobayashi K, Aoki M, Numaguchi Y, Shibuya M, Murohara T.
  • Journal Title: Arterioscler. Thromb. Vasc. Biol. 27 Pages: 99-105
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Angiotensin II Receptor Blocker Inhibits Neointimal Hyperplasia Through Regulation of Smooth Muscle-like Progenitor Cells. (2007)
  • Author(s): Yamada T, Kondo T, Numaguchi Y, Tsuzuki M, Matsubara T, Manabe I, Sata M, Nagai R, Murobara T.
  • Journal Title: Arterioscler. Thromb. Vasc. Biol. 27 Pages: 2363-2369
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Therapeutic angiogenesis using novel vascular endothelial growth factor-E/human placental growth factor chimera genes (2007)
  • Author(s): Inoue, N, Kondo, T, Kobayashi, K, Aoki, M, Numaguchi, Y, Shibuya, M, Murohara, T
  • Journal Title: Arterioscler. Thromb. Vasa Biol 27 Pages: 99-105
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Angiotensin II Receptor Blocker Inhibits Neointimal Hyperplasia Through Regulation of Smooth Muscle-like Progenitor Cells (2007)
  • Author(s): Yamada, T, Kondo, T, Numaguchi, Y, Tsuzuki, M, Matsubara, T, Manabe, I, Sata, M, Nagai, R, Murohara, T
  • Journal Title: Arterioscler. Thromb. Vase. Biol 27 Pages: 2363-2369
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Angiotensin II Receptor Blocker Inhibits Neointimal Hype-plasia Through Regulation of Smooth Muscle-like Progenitor Cells. (2007)
  • Author(s): Yamada T, Kondo T, Numaguchi Y, Tsuzuki M, Matsubara T, Manabe I, Sata M, Nagai R, Murohara T.
  • Journal Title: Arterioscler. Thromb. Vasc. Biol. 27 Pages: 2363-2369
  • Peer Reviewed
• [Journal Article] Therapeutic angiogenesis using novel vascular endothelial growth factor-E/human placental growth factor chimera genes. (2007)
  • Author(s): Inoue N, Kondo T, Kobayashi K, Aoki M, Numaguchi Y, Shibuya M, Murohara T
  • Journal Title: Arterioscler. Thromb. Vasc. Biol. 27(In press)
• [Journal Article] A strategy of retrograde injection of bone marrow mononuclear cells into the myocardium for the treatment of ischemic heart disease. (2006)
  • Author(s): Yokoyama S, Fukuda N, Li Y, Hagikura K, Takayama T, Kunimoto S, Honye J, Saito S, Wada M, Satomi A, Kato M, Mugishima H, Kusumi Y, Mitsumata M, Murohara T
  • Journal Title: J. Mol. Cell. Cardiol. 40 Pages: 24-34
• [Journal Article] Endothelial progenitor cell transplantation improves the outcome following liver injury in mice. (2006)
  • Author(s): Taniguchi E, Kin M, Torimura T, Nakamura T, Kumemura M, Hanada S, Hisamoto T, Yoshida T, Kawaguchi T, Baba S, Maeyama M, Koga H, Harada M, Kumashiro R, Ueno T, Ikeda H, Imaizumi T, Murohara I, Sata M
  • Journal Title: Gastroenterology 130 Pages: 521-531
• [Journal Article] Synergistic effect of combined intramyocardial CD34 cells and VEGF-2 gene therapy post-myocardial infarction. (2006)
  • Author(s): Shintani S, Kusano K, Ii M, Iwakura A, Heyd L, Curry C, Wecker A, Gavin M, Ma H, Keamey M, Silver M, Thome T, Murohara T, Losordo DW
  • Journal Title: Nat. Clin. Pract. Cardiovasc. Med. 3(Suppl.1)
• [Journal Article] The impact of the capability of circulating progenitor cell to differentiate on myocardial salvage in patients with primary acute myocardial infarction. (2006)
  • Author(s): Numaguchi Y, Sone T, Okumura K, Morita Y, Kubota R, Yokouchi K, Imai H, Harada M, Kondo T, Murohara T
  • Journal Title: Circulation 114(Suppl.1)
• [Journal Article] Combination therapy using angiopoietin-1 plasmid gene and autologous bone marrow cell implantation promotes functional angiogenesis. (2006)
  • Author(s): Kobayashi K, Kondo T, Inoue N, Aoki M, Mizuno M, Komori K, Yoshida J, Murohara T
  • Journal Title: Arterioscler. Thromb. Vasc. Biol. 26 Pages: 1465-1472
• [Presentation] Regulation of VEGF-mediated angiogenesis by the Akt/PKB substrate Girdin. (2007)
  • Author(s): Kitamura T, Asai N, Enomoto A, Maeda K, Murohara T, Takahashi M.
  • Organizer: American Heart Association, 2007
  • Place of Presentation: Orlando, USA
  • Year and Date: 2007-11-06
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] of VEGF-mediated angiogenesis by the Akt/PKB substrate Girdin (2007)
  • Author(s): Kitamura, T, Asai, N, Enomoto, A, Maeda, K, Murohara, T, Takahashi, M. Regulation
  • Organizer: American Heart Association 2007
  • Place of Presentation: Orlando, USA
  • Year and Date: 2007-11-06
  • Description: 「研究成果報告書概要(欧文)」より