Optimization of gene transfer conditions to adipose tissue and application toward hemophilia

• Project/Area Number: 17591007
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Hematology
• Research Institution: Jichi Medical University
• Principal Investigator: MIZUKAMI Hiroaki Jichi Medical University, School of Medicine, Assistant Professor, 医学部, 講師 (20311938)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2006: ¥1,500,000 (Direct Cost: ¥1,500,000); Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
• Keywords: adipose tissue / hemophilia / gene therapy / AAV Vectors / pluronic / erythropoietin / tissue specificity / polycythemia / 血清型 / 添加剤 / 凝固第IX因子 / 遺伝子導入 / 摘出手術

Research Abstract

Adipose tissue is potentially a suitable target of gene transfer for diseases like hemophilia. However, no efficient method has been developed for in vivo gene transfer. In this study, we explored the utility of excipients to augment gene transfer into the adipose tissue of mice and found that Pluronic F88 was useful for this purpose when combined with AAV serotype 1 vectors. The improvement was also demonstrated by vectors encoding murine erythropoietin, and the mice became polycythemic. Moreover, after removing transduced adipose tissue, plasma erythropoietin levels returned to normal, which suggests the unique advantage of this method. When AAV8-based vector was tested in the same manner, robust transgene expression was observed in the liver rather than the injected adipose tissue. This finding led to the discovery of ectopic liver expression following intramuscular injection of AAV8-based vectors. On the other hand, in experiments with vectors encoding coagulation factor IX gene, transgene expression level was not satisfactory in mice. The reason for this phenomenon is still unclear and need to be clarified in the future. Further optimization and analysis is necessary to improve the level of transgene expression and application to the diseases other than hemophilia.

Research Products

• [Journal Article] Prevention of diabetic retinopathy by intraocular soluble flt-1 gene transfer in a spontaneously diabetic rat model. (2007)
  • Author(s): Ideno J, et al.
  • Journal Title: Int J Mol Med 19巻 Pages: 75-79
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Suppression of ovarian cancer by muscle-mediated expression of soluble VEGFR-1/Flt-1 using adeno-associated virus serotype 1-derived vector. (2007)
  • Author(s): Takei Y, et al.
  • Journal Title: Int J Cancer 120巻 Pages: 278-284
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Adeno-associated virus-2 effectively transduces intrasynovial tenocytes with persistent expression of the transgene, but other serotypes do not. (2007)
  • Author(s): Wang X, et al.
  • Journal Title: Plast Reconstr Surg 119巻 Pages: 227-234
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Prevention of diabetic retinopathy by intraocular soluble flt-1 gene transfer in a spontaneously diabetic rat model. (2007)
  • Author(s): Ideno, J., Mizukami, H., Kakehashi, A., Saito, Y., Okada, T., Urabe, M., Kume, A., Kuroki, M., Kawakami, M., Ishibashi, S., Ozawa, K.
  • Journal Title: Int J Mol Med 19 Pages: 75-9
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Suppression of ovarian cancer by muscle-mediated expression of soluble VEGFR-1/Flt-1 using adeno-associated virus serotype 1-derived vector. (2007)
  • Author(s): Takei, Y., Mizukami, H., Saga, Y., Yoshimura, I., Hasumi, Y., Takayama, T., Kohno, T., Matsushita, T., Okada, T., Kume, A., Suzuki, M., Ozawa, K.
  • Journal Title: Int J Cancer 120 Pages: 278-84
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Tendon healing in vitro : Adeno-associated virus-2 effectively transduces intrasynovial tenocytes with persistent expression of the transgene, but other serotypes do not. (2007)
  • Author(s): Wang, X.T., Liu, P.Y., Tang, J.B., Mizukami, H., Xin, KQ., Ozawa, K., Ushijima, H.
  • Journal Title: Plast Reconstr Surg 119 Pages: 227-34
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Prevention of diabetic retinopathy by intraocular soluble flt-1 gene transfer in a spontaneously diabetic rat model. (2007)
  • Author(s): Ideno J, et al.
  • Journal Title: Int J Mol Med 19・1 Pages: 75-79
• [Journal Article] Suppression of ovarian cancer by muscle-mediated expression of soluble VEGFR-1/Flt-1 using adeno-associated virus serotype 1-derived vector. (2007)
  • Author(s): Takei Y, et al.
  • Journal Title: Int J Cancer 120・2 Pages: 278-84
• [Journal Article] Adeno-associated virus-2 effectively transduces intraynovial tenocytes with persistent expression of the transgene, but otherserotypes do not. (2007)
  • Author(s): Wang X, et al.
  • Journal Title: Plast Reconstr Surg 119・1 Pages: 227-34
• [Journal Article] Adipose tissue as a novel target for in vivo gene transfer by using adeno-associated virus vectors. (2006)
  • Author(s): Mizukami H, et al.
  • Journal Title: Hum Gene Ther 17巻 Pages: 921-928
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The induction of robust immune responses against HIV is supported by the inherent tropism of AAV5 for DC. (2006)
  • Author(s): Xin, KQ, et al.
  • Journal Title: J Virol 80巻 Pages: 11899-11910
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Utility of intraperitoneal administration as a route of AAV serotype 5 vector-mediated neonatal gene transfer. (2006)
  • Author(s): Ogura T, et al.
  • Journal Title: J Gene Med 8巻 Pages: 990-997
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Phenotype correction of hemophilia A mice with adeno-associated virus vectors carrying the B domain-deleted canine factor VIII gene. (2006)
  • Author(s): Ishiwata A, et al.
  • Journal Title: Thromb Res 118巻 Pages: 627-635
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A histone deacetylase inhibitor enhances recombinant adeno-associated virus-mediated gene expression in tumor cells. (2006)
  • Author(s): Okada T, et al.
  • Journal Title: Mol Ther 13巻 Pages: 738-746
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Removal of empty capsids from type 1 adeno-associated virus vector stocks by anion-exchange chromatography potentiates transgene expression. (2006)
  • Author(s): Urabe M, et al.
  • Journal Title: Mol Ther 13巻 Pages: 823-828
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Scalable generation of high-titer recombinant adeno-associated virus type 5 in insect cells. (2006)
  • Author(s): Urabe M, et al.
  • Journal Title: J Virol 80巻 Pages: 1874-1885
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Adipose tissue as a novel target for in vivo gene transfer by using adeno-associated virus vectors. (2006)
  • Author(s): Mizukami, H., Mimuro, J., Ogura, T., Okada, T., Urabe, M., Kume, A., Sakata, Y., Ozawa, K.
  • Journal Title: Hum Gene Ther 17 Pages: 921-8
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Utility of intraperitoneal administration as a route of AAV serotype 5 vector-mediated neonatal gene transfer. (2006)
  • Author(s): Ogura, T., Mizukami, H., Mimuro, J., Madoiwa, S., Okada, T., Matsushita, T., Urabe, M., Kume, A., Hamada, H., Yoshikawa, H., Sakata, Y., Ozawa, K.
  • Journal Title: J Gene Med 8 Pages: 990-7
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Phenotype correction of hemophilia A mice with adeno-associated virus vectors carrying the B domain-deleted canine factor VIII gene. (2006)
  • Author(s): Ishiwata, A., Mimuro, J., Kashiwakura, Y., Niimura, M., Takano, K., Ohmori, T., Madoiwa, S., Mizukami, H., Okada, T., Naka, H., Yoshioka, A., Ozawa, K., Sakata, Y.
  • Journal Title: Thromb Res 118 Pages: 627-35
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A histone deacetylase inhibitor enhances recombinant adeno-associated virus-mediated gene expression in tumor cells. (2006)
  • Author(s): Okada, T., Uchibori, R., Iwata-Okada, M., Takahashi, M., Nomoto, T., Nonaka-Sarukawa, M., Ito, T., Liu, Y., Mizukami, H., Kume, A., Kobayashi, E., Ozawa, K.
  • Journal Title: Mol Ther 13 Pages: 738-46
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Scalable generation of high-titer recombinant adeno-associated virus type 5 in insect cells. (2006)
  • Author(s): Urabe, M., Nakakura, T., Xin, KQ., Obara, Y., Mizukami, H., Kume, A., Kotin, RM., Ozawa, K.
  • Journal Title: J Virol 80 Pages: 1874-85
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Removal of empty capsids from type 1 adeno-associated virus vector stocks by anion-exchange chromatography potentiates transgene expression. (2006)
  • Author(s): Urabe, M., Xin, KQ., Obara, Y., Nakakura, T., Mizukami, H., Kume, A., Okuda, K., Ozawa, K.
  • Journal Title: Mol Ther 13 Pages: 823-8
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The induction of robust immune responses against HIV is supported by the inherent tropism of AAV5 for DC. (2006)
  • Author(s): Xin, KQ., Mizukami, H., Urabe, M., Toda, Y., Shinoda, K., Yoshida, A., Oomura, K., Kojima, Y., Ichino, M., Klinman, D., Ozawa K., Okuda, K.
  • Journal Title: J Virol 80 Pages: 11899-910
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Adipose tissue as a novel target for in vivo gene transfer by using adeno-associated virus vectors. (2006)
  • Author(s): Mizukami H, et al.
  • Journal Title: Hum Gene Ther 17・9 Pages: 921-8
• [Journal Article] The induction of robust immune responses against HIV is supported by the inherent tropism of AAV5 for DC. (2006)
  • Author(s): Xin, KQ, et al.
  • Journal Title: J Virol 80・24 Pages: 11899-910
• [Journal Article] Utility of intraperitoneal administraction as a route of AAV serotype 5 vector-mediated neonatal gene transfer. (2006)
  • Author(s): Ogura T, et al.
  • Journal Title: J Gene Med 8・8 Pages: 990-7
• [Journal Article] Phenotype correction of hemophilia A mice with adeno-associated virus vectors carrying the B domain-deleted canine factor VIII gene. (2006)
  • Author(s): Ishiwata A, et al.
  • Journal Title: Thromb Res 118・5 Pages: 627-35
• [Journal Article] A histone deacetylase inhibitor enhances recombinant adeno-associated virus-mediated gene expression in tumor cells. (2006)
  • Author(s): Okada T, et al.
  • Journal Title: Mol Ther 13・4 Pages: 738-46
• [Journal Article] Removal of empty capsids from type 1 adeno-associated virus vector stocks by anion-exchange chromatography potentiates transgene expression. (2006)
  • Author(s): Urabe M, et al.
  • Journal Title: Mol Ther 13・4 Pages: 823-8
• [Journal Article] Repair of articular cartilage defect by intraarticular administration of basic fibroblast growth factor gene, using adeno-associated virus vector. (2005)
  • Author(s): Hiraide A, et al.
  • Journal Title: Hum Gene Ther 16・12 Pages: 1413-1421
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Specific and efficient transduction of cochlear inner hair cells with recombinant adeno-associated virus type 3 vector. (2005)
  • Author(s): Liu Y, et al.
  • Journal Title: Mol Ther 12・4 Pages: 725-733
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Repair of articular cartilage defect by intraarticular administration of basic fibroblast growth factor gene, using adeno-associated virus vector. (2005)
  • Author(s): Hiraide, A., Yokoo, N., Xin, KQ., Okuda, K., Mizukami, H., Ozawa, K., Saito, T.
  • Journal Title: Hum Gene Ther 16 Pages: 1413-21
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Specific and efficient transduction of cochlear inner hair cells with recombinant adeno-associated virus type 3 vector. (2005)
  • Author(s): Liu, Y., Okada, T., Sheykholeslami, K., Shimazaki, K., Nomoto, T., Muramatsu, S., Kanazawa, T., Takeuchi, K., Ajalli, R., Mizukami, H., Kume, A., Ichimura, K., Ozawa, K.
  • Journal Title: Mol Ther 12 Pages: 725-33
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Repair of articular defect by intraarticular administration of basic fibroblast growth factor gene, using adeno-associated virus vector. (2005)
  • Author(s): Hiraide A, et al.
  • Journal Title: Hum Gene Ther 16・12 Pages: 1413-1421
• [Journal Article] Utility of intraperitoneal administration as a route of AAV serotype 5 vector-mediated neonatal gene transfer.
  • Author(s): Ogura T, et al.
  • Journal Title: J Gene Med (in press)
• [Journal Article] Removal of empty capsids from type 1 adeno-associated virus vector stocks by anion-exchange chromatography potentiates transgene expression.
  • Author(s): Urabe M, et al.
  • Journal Title: Mol Ther (in press)
• [Journal Article] Phenotype correction of hemophilia A mice with adeno-associated virus vectors carrying the B domain-deleted canine factor VIII gene.
  • Author(s): Ishiwata A, et al.
  • Journal Title: Thromb Res (in press)
• [Journal Article] A histone deacetylase inhibitor enhances recombinant adeno-associated virus-mediated gene expression in tumor cells.
  • Author(s): Okada T, et al.
  • Journal Title: Mol Ther (in press)
• [Book] 糖尿病学-基礎と臨床 (2007)
  • Author(s): 水上 浩明(分担執筆)
  • Total Pages: 1200
  • Publisher: 西村書店
  • Description: 「研究成果報告書概要(和文)」より