Regeneration of neurons and vessels using bone marrow cells and gene delivery

• Project/Area Number: 17300224
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied health science
• Research Institution: Osaka Medical College
• Principal Investigator: KUROIWA Toshihiko Osaka Medical College, Faculty of Medicine, Professor, 医学部, 教授 (30178115)
• Co-Investigator(Kenkyū-buntansha): MIYATAKE Shinichi Osaka Medical College, Faculty of Medicine, Associate Professor, 医学部, 助教授 (90209916) ; KAJIMOTO Yoshinaga Osaka Medical College, Faculty of Medicine, Assistant Professor, 医学部, 講師 (30224413) ; NONOGUCHI Naosuke Osaka Medical College, Faculty of Medicine, Research Associate, 医学部, 助手 (70388263) ; DEZAWA Mari Kyoto University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (50272323)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥6,000,000 (Direct Cost: ¥6,000,000); Fiscal Year 2006: ¥2,400,000 (Direct Cost: ¥2,400,000); Fiscal Year 2005: ¥3,600,000 (Direct Cost: ¥3,600,000)
• Keywords: Anti-aging / Senescence accelerated mouse / bone marrow cell / mesenchymal stromal cell / FGF / HGF / cerebral ischemia / 老化予防 / 脳梗塞治療 / 遺伝子導入 / 認知症 / 骨粗しょう症 / 老化 / 骨髄細胞移植 / 脳萎縮 / 老化促進マウス

Research Abstract

We preliminarily intravenously implanted bone marrow cells from young mouse to old senescence accelerated mouse (SAM). We evaluate the memory function by water maze, however, no improvement in memory function was observed. Therefore, intravenous administration of young bone marrow has little effect on preventing aging and further enhanced treatment such as useful gene induction and targeting administration to the brain or bone marrow.
Then we examined the effects of neurological improvement after transient middle cerebral artery occlusion (MCAO) in rats by a novel therapeutic strategy with FGF-2 gene-transferred mesenchymal stromal cells (MSCs) by the herpes simplex virus type 1 (HSV-1) vector. The stroke animals receiving FGF-2-modified MSCs demonstrated significant functional recovery compared with the other groups. Fourteen days after the MCAO, there was a significant reduction in infarction volume only in FGF-2-modified MSC-treated group. FGF-2 production in the FGF-2-modified MSC-t reated brain was significantly higher compared with the other groups at 3 and 7 days after MCAO. Administrated FGF-2-modified MSCs strongly expressed the FGF-2 protein, which was proven by ELISA. In conclusion these datum suggested that the FGF-2 gene-modified MSCs with the HSV-1 vector can contribute to remarkable functional recovery after stroke compared with MSCs transplantation alone.
The hepatocyte growth factor (HGF) is also one of the useful gene for tissue restoration. we introduce a new strategy combining MSCs and ex vivo HGF gene transferring with a multimutated HSV-1 vector in a rat transient MCAO model. The significant difference of infarction areas on day was detected only between the MSC-HGF group and the PBS group with the superacute treatment, but was detected among each group on day 14 with both transplantations. After the superacute transplantation, we detected abundant expression of HGF protein in the ischemic brain of the MSC-HGF group compared with others on day 1 after treatment, and it was maintained for at least 2 weeks. Furthermore, we determined that the increased expression of HGF was derived from the transferred HGF gene in gene-modified MSCs. The percentage of apoptosis-positive cells in the ischemic boundary zone (IBZ) was significantly decreased, while that of remaining neurons in the cortex of the IBZ was significantly increased in the MSC-HGF group compared with others. The present study shows that combined therapy is more therapeutically efficient than MSC cell therapy alone, and it may extend the therapeutic time window from superacute to acute phase.
Following these experiments, the improving effect is mainly due to anti-apoptotic and neuron-protective effects and there were no evidence for neuron-generation. Another effective methods such as intra-bone marrow injection of bone marrow cell should be examined in the future.

Research Products

• [Journal Article] Novel Therapeutic Strategy for Stroke in Rats by Bone Marrow Stromal Cells and Ex Vivo HGF Gene Transfer with HSV-1 Vector (2006)
  • Author(s): M Zhao, N Nonoguchi, N Ikeda, T Watanabe, D Furutama, D Miyazawa, H Funakoshi, Y Kajimoto, T Nakamura, M Dezawa, M Shibata, Y Otsuki, RS. Coffin, W Liu, T Kuroiwa, S Miyatake
  • Journal Title: J Cereb Blood Flow Metab 26・9 Pages: 1176-1188
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Novel Therapeutic Strategy for Stroke in Rats by Bone Marrow Stromal Cells and Ex Vivo HGF Gene Transfer with HSV-1 Vector (2006)
  • Author(s): M Zhao, N Nonoguchi, N Ikeda, T Watanabe, D Furutama, D Miyazawa, H Funakoshi, Y Kajimoto, T Nakamura, M Dezawa, M Shibata, Y Otsuki, RS. Coffin, W Liu, T Kuroiwa, S Miyatake
  • Journal Title: J Cereb Blood Flow Metab 26 (9) Pages: 1176-1188
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Novel Therapeutic Strategy for Stroke in Rats by Bone Marrow Stromal Cells and Ex Vivo HGF Gene Transfer with HSV-1 Vector (2006)
  • Author(s): M Zhao, N Nonoguchi, N Ikeda, T Watanabe, D Furutama, D Miyazawa, H Funakoshi, Y Kajimoto, T Nakamura, M Dezawa, M Shibata, Y Otsuki, RS.Coffin, W Liu, S Miyatake
  • Journal Title: J Cereb Blood Flow Metab 26・9 Pages: 1176-1188
• [Journal Article] Bone Marrow Stromal Cells That Enhanced FGF-2 Secretion by Herpes Simplex Virus Vector Improve Neurological Outcome after Transient Focal Cerebral Ischemia in Rats. (2005)
  • Author(s): Ikeda N, Nonoguchi N, Zhao MZ, Watanabe T, Kajimoto Y, Furutama D, Kimura F, Dezawa M, Robert S, Otsuki Y, Kuroiwa T, Miyatake SI
  • Journal Title: Stroke 36・12 Pages: 2725-2730
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Bone Marrow Stromal Cells That Enhanced FGF-2 Secretion by Herpes Simplex Virus Vector Improve Neurological Outcome after Transient Focal Cerebral Ischemia in Rats. (2005)
  • Author(s): Ikeda N, Nonoguchi N, Zhao MZ, Watanabe T, Kajimoto Y, Furutama D, Kimura F, Dezawa M, Robert S, Otsuki Y, Kuroiwa T, Miyatake SI
  • Journal Title: Stroke 36 (12) Pages: 2725-2730
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Bone Marrow Stromal Cells That Enhanced FGF-2 Secretion by Herpes Simplex Virus Vector Improve Neurological Outcome after Transient Focal Cerebral Ischemia in Rats. (2005)
  • Author(s): Ikeda N, noguchi N, Zha MZ, Watanabe T, Kajimoto Y, Furutama D, Kimura F, Dezawa M, Robert S, Otsuki Y, Kutoiwa T, Miyatake SI
  • Journal Title: Stroke 36・12 Pages: 2725-2730