Development of new gene-repair therapy for primary immunodeficiencies with artificial nuclease

• Project/Area Number: 15K15393
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Pediatrics
• Research Institution: Fukuoka Children’s Hospital (2017); Kyushu University (2015-2016)
• Principal Investigator: HARA Toshiro 地方独立行政法人福岡市立病院機構福岡市立こども病院(臨床研究部), 臨床研究部, 院長 (40150445)
• Co-Investigator(Kenkyū-buntansha): 高田 英俊 九州大学, 医学研究院, 教授 (70294931) ; 山元 裕之 九州大学, 環境発達医学研究センター, 特任助教 (00710170)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 遺伝子治療 / 人工ヌクレアーゼ

Outline of Final Research Achievements

To examine the possibility of using gene therapy for primary immunodeficiencies, we constructed a helper-dependent adenovirus/adeno-associated virus hybrid vector to serve as a donor template, and a sequence-specific gRNA for double strand breaks. Human cord blood CD34 positive cells were infected with the hybrid vector and electroporated with gRNA and Cas9 protein. This delivery method can be a new gene-repair therapy for clinical application.

Research Products

• [Journal Article] BTK gene targeting by homologous recombination using a helper-dependent adenovirus/adeno-associated virus hybrid vector. (2016)
  • Author(s): Yamamoto H, Ishimura M, Ochiai M, Takada H, Kusuhara K, Nakatsu Y, Tsuzuki T, Mitani K, Hara T.
  • Journal Title: Gene Ther Volume: 23 Issue: 2 Pages: 205-13
  • DOI: 10.1038/gt.2015.91
  • Peer Reviewed / Open Access
• [Presentation] BTK Gene Targeting by Homologous Recombination Using a Helper-Dependent Adenovirus/Adeno-Associated Virus Hybrid Vector and CRISPR/Cas9 System (2017)
  • Author(s): Hiroyuki Yamamoto, Hidetoshi Takada, Masataka Ishimura, Masayuki Ochiai, Toshiro Hara, Shouichi Ohga
  • Organizer: The 8th JSH International Symposium 2017
  • Int'l Joint Research
• [Presentation] BTK gene targeting by homologous recombination using a helper-dependent adenovirus/adeno-associated virus hybrid vector (2016)
  • Author(s): 山元裕之
  • Organizer: 第２７回福岡国際母子総合研究シンポジウム
  • Place of Presentation: JR 博多シティ（福岡県福岡市）
  • Year and Date: 2016-08-28
  • Int'l Joint Research
• [Presentation] Gene Therapy for PID (2015)
  • Author(s): 高田英俊
  • Organizer: The 11th Congress of Asian Society for Pediatric Research (ASPR 2015)
  • Place of Presentation: 大阪府立国際会議場（大阪府大阪市）
  • Year and Date: 2015-04-15
  • Int'l Joint Research