Development of a site-specific gene insertion technology for regenerative medicine:Basic study using developmental engineering

• Project/Area Number: 23659493
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Hematology
• Research Institution: Jichi Medical University
• Principal Investigator: OZAWA Keiya 自治医科大学, 医学部, 教授 (30137707)
• Co-Investigator(Kenkyū-buntansha): URABE Masashi 自治医科大学, 医学部, 講師 (40213516)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2011: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 再生医療 / 幹細胞 / iPS細胞 / 遺伝子導入 / 遺伝子組込み

Research Abstract

We developed a site-specific gene insertion technology for stem cells by using an integration machinery based on adeno-associated virus (AAV), which preferentially integrates into the AAVS1 site on chromosome 19. We tested this system with a GFP plasmid

Research Products

• [Journal Article] Minimizing the inhibitory effect of neutralizing antibody for efficient gene expression in the liver with adeno-associated virus 8 vectors. (2013)
  • Author(s): Mimuro J, Mizukami H, Hishikawa S, Ikemoto T, Ishiwata A, Sakata A, Ohmori T, Madoiwa S, Ono F, Ozawa K, and Sakata Y
  • Journal Title: Mol Ther Volume: 21 Issue: 2 Pages: 318-323
  • DOI: 10.1038/mt.2012.258
  • Peer Reviewed
• [Journal Article] NF-κB activity regulates mesenchymal stem cell accumulation at tumor sites. (2013)
  • Author(s): Uchibori R, Tsukahara T, Mizuguchi H, Saga Y, Urabe M, Mizukami H, Kume A, Ozawa K
  • Journal Title: Cancer Research Volume: 73 Issue: 1 Pages: 364-372
  • DOI: 10.1158/0008-5472.can-12-0088
  • Peer Reviewed
• [Journal Article] Downregulation of indoleamine-2, 3-dioxygenase in cervical cancer cells suppresses tumor growth by promoting natural killer cell accumulation. (2012)
  • Author(s): Sato N, Saga Y, Mizukami H, Wang D, Takahashi S, Nonaka H, Fujiwara H, Takei Y, Machida S, Takikawa O, Ozawa K, Suzuki M.
  • Journal Title: Oncol Rep. Volume: 28 Pages: 1574-1578
  • Peer Reviewed
• [Journal Article] Intra-articular injection of mesenchymal stem cells expressing coagulation factorameliorates hemophilic arthropathy in factor VIII-deficient mice. (2012)
  • Author(s): Kashiwakura, Y
  • Journal Title: J Thromb Haemost. Volume: 10(9) Issue: 9 Pages: 1802-1813
  • DOI: 10.1111/j.1538-7836.2012.04851.x
  • Peer Reviewed
• [Journal Article] Cetuximab inhibits the growth of mucinous ovarian carcinoma tumor cells lacking KRAS gene mutations. (2012)
  • Author(s): Sato, N.
  • Journal Title: Oncol Rep. Volume: 27 Pages: 1336-1340
  • DOI: 10.3892/or.2012.1626
  • Peer Reviewed
• [Journal Article] Interleukin-10 expression induced by adeno-associated virus vector suppresses proteinuria in Zucker obese rats (2012)
  • Author(s): Ogura M, Urabe M, Akimoto T, Onishi A, Ito C, Ito T, Tsukahara T, Mizukami H, Kume A, Muto S, Kusano E, and Ozawa K.
  • Journal Title: Gene Ther Volume: 19巻 Issue: 5 Pages: 476-82
  • DOI: 10.1038/gt.2011.183
  • Peer Reviewed
• [Journal Article] Recovery of neurogenic amines in phenylketonuria mice after liver-targeted gene therapy (2012)
  • Author(s): Yagi, H., Sanechika, S., Ichinose, H., Sumi-Ichinose, C., Mizukami, H., Urabe, M., Ozawa, K., and Kume, A.
  • Journal Title: Neuroreport Volume: 23 Issue: 1 Pages: 30-34
  • DOI: 10.1097/wnr.0b013e32834e3a87
  • Peer Reviewed
• [Journal Article] Indoleamine-2,3-dioxygenase, an immunosuppressive enzyme that inhibits natural killer cell function, as a useful target for ovarian cancer therapy (2012)
  • Author(s): Wang, D.
  • Journal Title: Int J Oncol Volume: 40 Issue: 4 Pages: 929-34
  • DOI: 10.3892/ijo.2011.1295
  • Peer Reviewed
• [Journal Article] Morphologic analysis in myelodysplastic syndromes with del(5q) treated with lenalidomide. A Japanese multiinstitutional study (2012)
  • Author(s): Matsuda A, Taniwaki M, Jinnai I, Harada H, Watanabe M, Suzuki K, Yanagita S, Suzuki T, Yoshida Y, Kimura A, Tsudo M, Tohyama K, Takatoku M, Ozawa K
  • Journal Title: Leuk Res Volume: (in press) Issue: 5 Pages: 575-580
  • DOI: 10.1016/j.leukres.2011.11.011
  • Peer Reviewed
• [Journal Article] Downregulation of indoleamine-2,3-dioxygenase in cervical cancer cells suppresses tumor growth by promoting natural killer cell accumulation. (2012)
  • Author(s): Sato, N.
  • Journal Title: Oncol Rep. Volume: 28 Issue: 5 Pages: 1574-1578
  • DOI: 10.3892/or.2012.1984
  • Peer Reviewed
• [Journal Article] Morphologic analysis in myelodysplastic syndromes with del(5q) treated with lenalidomide. A Japanese multiinstitutional study. (2012)
  • Author(s): Matsuda, A.
  • Journal Title: Leuk Res. Volume: 36 Pages: 575-80
  • Peer Reviewed
• [Journal Article] Recov ery of neurogenic amines in phenylketonuria mice after liver-targeted gene therapy. (2012)
  • Author(s): Yagi, H.
  • Journal Title: Neuroreport. Volume: 23 Pages: 30-4
  • Peer Reviewed
• [Journal Article] Vincristine potentiates the anti- proliferative effect of an aurora kinase inhibitor, VE-465, in myeloid leukemia cells (2011)
  • Author(s): Yoshida K, Nagai T, Ohmine K, Uesawa M, Sripayap P, Ishida Y, and Ozawa K.
  • Journal Title: Biochem Pharmacol Volume: 82 Issue: 12 Pages: 1884-1890
  • DOI: 10.1016/j.bcp.2011.09.015
  • Peer Reviewed
• [Journal Article] Development of a mouse model for lymph node metastasis with endometrial cancer (2011)
  • Author(s): Takahashi K, Saga Y, Mizukami H, Takei Y, Urabe M, Kume A, Suzuki M, and Ozawa K
  • Journal Title: Cancer Sci Volume: 102巻 Issue: 12 Pages: 2272-7
  • DOI: 10.1111/j.1349-7006.2011.02099.x
  • NAID: 10030246395
  • Peer Reviewed
• [Journal Article] Selective optical control of synaptic transmission in the subcorticalvisual pathway by activation of viral vector-expressed halorhodopsin (2011)
  • Author(s): Kaneda K, Kasahara H, Matsui R, Katoh T, Mizukami H, Ozawa K, Watanabe D, Isa T
  • Journal Title: PLoSONE Volume: 6 Issue: 4 Pages: 18452-18452
  • DOI: 10.1371/journal.pone.0018452
  • Peer Reviewed
• [Journal Article] Lack of IL-21 signal attenuates graft-versus-leukemia effect in the absence of CD8 T-cells (2011)
  • Author(s): Meguro A, Ozaki K, Hatanaka K, Oh I, Sudo K, Ohmori T, Matsu H, Tatara., Sato K, Sakata Y, Nakae S, Leonard W J, and Ozawa K
  • Journal Title: Bone Marrow Transplant Volume: 46巻 Issue: 12 Pages: 1557-1565
  • DOI: 10.1038/bmt.2010.342
  • Peer Reviewed
• [Journal Article] Mesenchymal stromal cells inhibit Th17 but not regulatory T-cell differentiation (2011)
  • Author(s): Tatara R, Ozaki K, Kikuchi Y, Hatanaka K, Oh I, Meguro A, Matsu H, Sato K, Ozawa K.
  • Journal Title: Cytotherapy Volume: 13 Issue: 6 Pages: 686-694
  • DOI: 10.3109/14653249.2010.542456
  • Peer Reviewed
• [Journal Article] Complete restoration of phenylalanine oxidation in phenylketonuria mouse by a self-complementary adeno-associated virus vector (2011)
  • Author(s): Yagi H, Ogura T, Mizukami H, Urabe M, Hamada H, Yoshikawa H, Ozawa K and Kume A
  • Journal Title: J. Gene Med Volume: 13巻 Issue: 2 Pages: 114-122
  • DOI: 10.1002/jgm.1543
  • Peer Reviewed
• [Journal Article] Vincristine potentiates the anti-proliferative effect of an aurora kinase inhibitor, VE-465, in myeloid leukemia cells. (2011)
  • Author(s): Yoshida, K.
  • Journal Title: Biochem Pharmacol. Volume: 82 Pages: 1884-1890
  • Peer Reviewed
• [Journal Article] Selective optical control of synaptic transmission in the subcortical visual pathway by activation of viral vector-expressed halorhodopsin. (2011)
  • Author(s): Kaneda, K.
  • Journal Title: PLoS One. Volume: 6
  • Peer Reviewed
• [Presentation] Dose-response relationship of factor IX expression in non-human primates following IV administration of AAV vectors (2012)
  • Author(s): Mizukami H
  • Organizer: The 15th Annual Meeeting of The American Society of Gene and Cell Therapy
  • Place of Presentation: Pennsylvania Convention Center (Philadelphia, USA))
• [Presentation] AAV-vector mediated systemic delivery of interleukin-10 attenuates the progression of peritoneal fibrosis in a rat model of peritoneal dialysis (2012)
  • Author(s): Onishi A
  • Organizer: The 15th Annual Meeeting of The American Society of Gene and Cell Therapy
  • Place of Presentation: Pennsylvania Convention Center (Philadelphia, USA)
• [Presentation] Evaluation of anti-tumor effects mediated by engineered T lymphocytes expressing a CD19 specific CAR for B cell lynphoma. (2012)
  • Author(s): Tsukahara T
  • Organizer: The 15th Annual Meeeting of The American Society of Gene and Cell Therapy
  • Place of Presentation: Pennsylvania Convention Center (Philadelphia, USA)
• [Presentation] Site-specific gene insertion of transgene by adeno-associated virus integration machinery (2012)
  • Author(s): Urabe M
  • Organizer: The 10th Annual Meeting of The International Society for Stem Cell Research
  • Place of Presentation: Pacifico Yokohama (Yokohama)
• [Presentation] Relationship between vector dose and factor IX expression in non-human primates following administration of AAV8 vectors (2012)
  • Author(s): Mizukami H
  • Organizer: 第18回日本遺伝子治療学会学術集会
  • Place of Presentation: 熊本テルサ(熊本)
• [Presentation] AADC gene therapy for parkinson disease: long term outcomes and preliminary results of an autopsy case (2012)
  • Author(s): Muramatsu S
  • Organizer: 第18回日本遺伝子治療学会学術集会
  • Place of Presentation: 熊本テルサ(熊本)
• [Presentation] Interleukin-10 prevents the progression of peritoneal fibrosis in an animal model of peritoneal dialysis (2012)
  • Author(s): Onishi A
  • Organizer: 第18回日本遺伝子治療学会学術集会
  • Place of Presentation: 熊本テルサ(熊本)
• [Presentation] Genetically modified T lymphocytes targeted to CD19 mediated-cytotoxicity for B cell lymphoma (2012)
  • Author(s): Tsukahara T
  • Organizer: 第18回日本遺伝子治療学会学術集会
  • Place of Presentation: 熊本テルサ(熊本)
• [Presentation] Manipulationg the genome in stem cells. (2012)
  • Author(s): Urabe M
  • Organizer: 第18回日本遺伝子治療学会学術集会
  • Place of Presentation: 熊本テルサ(熊本)
• [Presentation] Anti-tumor effects of anti-CD19 chimeric antigen receptor modified T-cells for B cell lymphoma. (2012)
  • Author(s): Tsukahara T
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌(札幌)
• [Presentation] A novel anti-tumor mechanism of galanin receptor type 2 in head and neck qquamous cell carcinoma cells (2012)
  • Author(s): Uehara T
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌(札幌)
• [Presentation] Dose-response relationship of factor IX expression in non-human primates following IV administration of AAV vectors (2012)
  • Author(s): Mizukami, H.
  • Organizer: The 15th Annual Meeeting of The American Society of Gene and Cell Therapy
  • Place of Presentation: Philadelphia Convention Center (Philadelphia,USA)
• [Presentation] AAV-vector mediated systemic delivery of interleukin-10 attenuates the progression of peritoneal fibrosis in a rat model of peritoneal dialysis (2012)
  • Author(s): Onishi, A.
  • Organizer: The 15th Annual Meeeting of The American Society of Gene and Cell Therapy
  • Place of Presentation: Philadelphia Convention Center (Philadelphia,USA)
• [Presentation] Evaluation of anti-tumor effects mediated by engineered T lymphocytes expressing a CD19 specific CAR for B cell lynphoma (2012)
  • Author(s): Tsukahara, T.
  • Organizer: The 15th Annual Meeeting of The American Society of Gene and Cell Therapy
  • Place of Presentation: Philadelphia Convention Center (Philadelphia,USA)
• [Presentation] Site-specific gene insertion of transgene by adeno-associated virus integration machinery (2012)
  • Author(s): Urabe, M.
  • Organizer: The 10th Annual Meeting of The International Society for Stem Cell Research
  • Place of Presentation: パシフィコ横浜（横浜）
• [Presentation] Relationship between vector dose and factor IX expression in non-human primates following administration of AAV8 vectors (2012)
  • Author(s): Mizukami, H.
  • Organizer: 第18回日本遺伝子治療学会学術集会
  • Place of Presentation: 熊本テルサ（熊本）
• [Presentation] AADC gene therapy for parkinson disease: long term outcomes and preliminary results of an autopsy case (2012)
  • Author(s): Muramatsu, S.
  • Organizer: 第18回日本遺伝子治療学会学術集会
  • Place of Presentation: 熊本テルサ（熊本）
• [Presentation] Interleukin-10 prevents the progression of peritoneal fibrosis in an animal model of peritoneal dialysis (2012)
  • Author(s): Onishi, A.
  • Organizer: 第18回日本遺伝子治療学会学術集会
  • Place of Presentation: 熊本テルサ（熊本）
• [Presentation] Genetically modified T lymphocytes targeted to CD19 mediated-cytotoxicity for B cell lymphoma (2012)
  • Author(s): Tsukahara, T.
  • Organizer: 第18回日本遺伝子治療学会学術集会
  • Place of Presentation: 熊本テルサ（熊本）
• [Presentation] Manipulationg the genome in stem cells (2012)
  • Author(s): Urabe, M.
  • Organizer: 第18回日本遺伝子治療学会学術集会
  • Place of Presentation: 熊本テルサ（熊本）
• [Presentation] Anti-tumor effects of anti-CD19 chimeric antigen receptor modified T-cells for B cell lymphoma (2012)
  • Author(s): Tsukahara, T.
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌（札幌）
• [Presentation] A novel anti-tumor mechanism of galanin receptor type 2 in head and neck qquamous cell carcinoma cells (2012)
  • Author(s): Uehara, T.
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌（札幌）
• [Presentation] The adeno-associated virus p5 promoter sequence mediated integration of transgene into the AAVS1 site (2011)
  • Author(s): Urabe M
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜)
  • Year and Date: 2011-12-13
• [Presentation] The p5 promoter of adeno- associated virus for AAVS1-specific integration. (2011)
  • Author(s): Urabe M
  • Organizer: The XVth International Congress of Virology
  • Place of Presentation: 札幌コンベンションセンター(札幌)
  • Year and Date: 2011-09-13
• [Presentation] Relationship between neutralizing Antibody and factor IX expression in non-human primates following iv administration of AAV8 vectors (2011)
  • Author(s): Mizukami H
  • Organizer: The XVth International Congress of Virology
  • Place of Presentation: 札幌コンベンションセンター(札幌)
  • Year and Date: 2011-09-13
• [Presentation] The role of the p5 promoter of adeno-associated virus in AAVS1-directed integration. (2011)
  • Author(s): Urabe M
  • Organizer: The 17th Annual Meeting of the Japan Society of Gene Therapy
  • Place of Presentation: 九州大学医学部百年講堂(福岡)
  • Year and Date: 2011-07-17
• [Presentation] The p5 promoter sequence of AAV genome regulates the direction of transgene insertion into the AAVS1 site. (2011)
  • Author(s): Urabe M
  • Organizer: The 14th Annual Meeting of the American Society of Gene and Cell Therapy
  • Place of Presentation: The Washington State Convention Center (Seattle, USA)
  • Year and Date: 2011-05-21
• [Presentation] Relationship between neutralizing Antibody and factor IX expression in non-human primates following iv administration of AAV8 vectors (2011)
  • Author(s): Mizukami H
  • Organizer: The 14th Annual Meeting of the American Society of Gene and Cell Therapy
  • Place of Presentation: The Washington State Convention Center (Seattle, USA)
  • Year and Date: 2011-05-19
• [Presentation] Relationship between neutralizing antibody and factor IX expression in non-human primates following iv administration of AAV8 vectors.
  • Author(s): Mizukami, H.
  • Organizer: The 14th Annual Meeting of the American Society of Gene and Cell Therapy
  • Place of Presentation: The Washington State Convention Center（Seattle）
• [Presentation] The p5 promoter sequence of AAV genome regulates the direction of transgene insertion into the AAVS1 site.
  • Author(s): Urabe, M.
  • Organizer: The 14th Annual Meeting of the American Society of Gene and Cell Therapy
  • Place of Presentation: The Washington State Convention Center（Seattle）
• [Presentation] The role of the p5 promoter of adeno-associated virus in AAVS1-directed integration.
  • Author(s): Urabe, M.
  • Organizer: 第17回日本遺伝子治療学会学術集会
  • Place of Presentation: 九州大学医学部 百年講堂(福岡 ）
• [Presentation] The p5 promoter of adeno-associated virus for AAVS1-specific integration.
  • Author(s): Urabe, M.
  • Organizer: The XVth International Congress of Virology
  • Place of Presentation: 札幌コンベンションセンター（札幌）
• [Presentation] Relationship between neutralizing antibody and transgene expression in non-human primates following administration of AAV8 vectors.
  • Author(s): Mizukami, H.
  • Organizer: The XVth International Congress of Virology
  • Place of Presentation: 札幌コンベンションセンター（札幌）
• [Presentation] The adeno-associated virus p5 promoter sequence mediated integration of transgene into the AAVS1 site.
  • Author(s): Urabe, M.
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（横浜）