Therapeutic angiogenesis using noble human VEGF-E chimeric genes and autologous bone marrow mononuclear cells

• Project/Area Number: 16390221
• 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, 大学院・医学系研究科, 教授 (90299503)
• Co-Investigator(Kenkyū-buntansha): KONDO Takahisa Nagoya University, University Hospital, Assistant Professor, 医学部附属病院, 講師 (00303644) ; NUMAGUCHI Yasushi Nagoya University, School of Medicine, Associate Professor, 医学部, 助教授 (90378224) ; KOMORI Kimihiro Nagoya University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (40225587) ; SHIBUYA Masafumi University of Tokyo, Institute of Medical Science, Professor, 医科学研究所, 教授 (10107427)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥14,300,000 (Direct Cost: ¥14,300,000); Fiscal Year 2005: ¥6,900,000 (Direct Cost: ¥6,900,000); Fiscal Year 2004: ¥7,400,000 (Direct Cost: ¥7,400,000)
• Keywords: Therapeutic angiogenesis / Vascular regeneration / Regenerative medicine / Gene therapy / Growth factor / Bone marrow cells / Stem cells / Endothelial progenitor cells / 血管再生 / 血管成長因子 / 再生医療 / 虚血性心疾患 / 血管透過性 / 炎症反応

Research Abstract

Background
Vascular endothelial growth factor-A (VEGF-A) promotes angiogenesis but causes adverse side effects such as edema or tissue inflammation. VEGF-E, found in the genome of the Orf virus, specifically binds to VEGF receptor-2 and shows mitotic activity on endothelial cells. Recently, we created two forms of VEGF-E and human placental growth factor (P1GF) chimera genes (VEGF-E chimera #9 and VEGF-E chimera #33), which are humanized genes holding VEGF-E function but less antigenicity.
Methods and Results
We examined potential proangiogenic activities of these chimera genes. Four types of expression plasmids (pCDNA3.1-LacZ, phVEGF-A, pVEGF-Echimera#9 and pVEGF-Echimera#33, n=8 each) were administered in a rat model of hindlimb ischemia. Either pVEGF-E chimera #9, pVEGF-E chimera #33 or phVEGF-A significantly increased the ratio of ischemic/normal hindlimb blood-flow compared to the control pCDNA3.1-LacZ treated group. Histochemical staining by alkaline phosphatase also revealed that either pVEGF-Echimera#9, pVEGF-Echimera#33 or phVEGF-A increased the capillary density compared to the pCDNA3.1-LacZ treated group. Furthermore, immunostaining for anti-ED1 revealed that lower number of macrophages were infiltrated in both pVEGF-Echimera#9 and pVEGF-Echimera#33 groups compared to the phVEGF-A group.
Conclusions
Novel VEGF-E/human P1GF chimera genes, pVEGF-Echimera#9 and pVEGF-Echimera#33, significantly stimulated angiogenesis in response to tissue ischemia, whose extent was almost identical to that induced by phVEGF-A with less tissue inflammation responses.

Research Products

• [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, Kearncy M, Silver M, Thorne T, Murohara T, Losordo DW
  • Journal Title: Nat. Clin. Pract. Cardiovasc. Med. 3
  • Description: 「研究成果報告書概要(和文)」より
• [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 (In press)
  • Description: 「研究成果報告書概要(和文)」より
• [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, Kearney M, Silver M, Thorne T, Murohara T, Losordo DW
  • Journal Title: Nat.Clin.Pract.Cardiovasc.Med. 3
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Combination therapy using angiopoietin-1 plasmid gene and autologous bone marrow cell implantation promotes functional anigiogenesis. (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. (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [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] 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, Kearney M, Silver M, Thorne T, Murohara T, Losordo DW.
  • Journal Title: Nat.Clin.Pract.Cardiovasc.Med. 3
• [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. (In press)
• [Journal Article] Therapeutic neovascularization using cord blood-derived endothelial progenitor cells for diabetic neuropathy. (2005)
  • Author(s): Naruse K, Hamada Y, Nakashima E, Kato K, Mizubayashi R, Kamiya H, Yuzawa Y, Matsuo S, Murohara T, Matsubara T, Oiso Y, Nakamura J
  • Journal Title: Diabete 54 Pages: 1823-1828
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Therapeutic angiogenesis with intramuscular injection of low dose recombinant granulocyte-colony stimulating factor. (2005)
  • Author(s): Lee M, Aoki M, Kondo T, Okumura K, Komori K, Murohara T
  • Journal Title: Arterioscler. Thromb. Vasc. Biol 25 Pages: 2535-2541
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Therapeutic neovascularization using cord blood-derived endothelial progenitor cells for diabetic neuropathy. (2005)
  • Author(s): Naruse K, Hamada Y, Nakashima E, Kato K, Mizubayashi R, Kamiya H, Yuzawa Y, Matsuo S, Murohara T, Matsubara T, Oiso Y, Nakamura J.
  • Journal Title: Diabetes. 54 Pages: 1823-1828
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Therapeutic angiogenesis with intramuscular injection of low dose recombinant granulocyte-colony stimulating factor. (2005)
  • Author(s): Lee M, Aoki M, Kondo T, Okumura K, Komori K, Murohara T.
  • Journal Title: Arterioscler.Thromb.Vasc.Biol. 25 Pages: 2535-2541
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] CD34+ cells home, proliferate and participate in capillary formation and in combination with CD34- cells enhance tube formation in a 3D matrix. (2005)
  • Author(s): Rookmaaker MB, Verhaar MC, Loomans CJM, Peters E, Morohara T, Staal FJT, van Zonneveld AJ, Koolwijk P, Rabelink TJ, van Hinsbergh VWM.
  • Journal Title: Arterioscler.Thromb.Vasc.Biol 25 Pages: 1843-1850
• [Journal Article] Therapeutic angiogenesis with intramuscular injection of low does recombinant granulocyte-colony stimulating factor. (2005)
  • Author(s): Lee M, Aoki M, Kondo T, Okumura K, Komori K, Murohara T.
  • Journal Title: Arterioscler.Thromb.Vasc.Biol. 25 Pages: 2535-2541
• [Journal Article] Autologous bone-marrow mononuclear cell implantation improves endothelium-dependent vasodilation in patients with limb ischemia. (2004)
  • Author(s): Higashi Y, Kimura M, Hara K, Noma K, Jituiki D, Nakagawa K, Oshima T, Chayama K, Goto C, Matsubara H, Murohara T, Yoshizumi M
  • Journal Title: Circulation 109 Pages: 1215-1218
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Smoking cessation rapidly increases endothelial progenitor cells in peripheral blood in chronic smokers. (2004)
  • Author(s): Kondo T, Hayashi M, Kinoshita K, Kobayashi K, Iino S, Inden Y, Murohara T
  • Journal Title: Arterioscler. Thromb. Vasc. Biol. 24 Pages: 1442-1447
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Autologous bone-marrow mononuclear cell implantation improves endothelium-dependent vasodilation in patients with limb ischemia. (2004)
  • Author(s): Higashi Y, Kimura M, Hara K, Noma K, Jituiki D, Nakagawa K, Oshima T, Chayama K, Goto C, Matsubara H, Murohara T, Yoshizumi M
  • Journal Title: Circulation. 109 Pages: 1215-1218
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Smoking cessation rapidly increases endothelial progenitor cells in peripheral blood in chronic smokers. (2004)
  • Author(s): Kondo T, Hayashi M, Kinoshita K, Kobayashi K, Iino S, Inden Y, Murohara T.
  • Journal Title: Arterioscler.Thromb.Vasc.Biol. 24 Pages: 1442-1447
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Catheter-based prostacyclin synthase gene transfer prevents in-stent restenosis in rabbit atheromatous arteries. (2004)
  • Author(s): Numaguchi, Murohara et al.
  • Journal Title: Cardiovascular Research 61 Pages: 177-185
• [Journal Article] Autologous bone-marrow mononuclear cell implantation improves endothelium-dependent vasodilation in patients with limb ischemia (2004)
  • Author(s): Higashi, Murohara et al.
  • Journal Title: Circulation 109 Pages: 1215-1218
• [Journal Article] Smoking cessation rapidly increases endothelial progenitor cells in peripheral blood in chronic smokers (2004)
  • Author(s): Kondo, Murohara et al.
  • Journal Title: Arteriosclerosis Thrombosis Vascular Biology 24 Pages: 1442-1447
• [Journal Article] Inhibition of platelet adherence to mononuclear cells by α-tocopherol : Role of P-selectin (2004)
  • Author(s): Murohara et al.
  • Journal Title: Circulation 110 Pages: 141-148
• [Journal Article] Angiogenesis and vasculogenesis are impaired in precocious aging Klotho mouse. (2004)
  • Author(s): Shimada, Murohara et al.
  • Journal Title: Circulation 110 Pages: 1148-1155
• [Journal Article] Augmentation of therapeutic angiogenesis using genetically-modified human endothelial progenitor cells with altered glycogen synthase kinase-3b activity. (2004)
  • Author(s): Choi, Murohara et al.
  • Journal Title: Journal of Biological Chemistry 279 Pages: 49430-49438