Development of gene repair therapy using autologous cells for primary immunodeficiencies

• Project/Area Number: 22249043
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Kyushu University
• Principal Investigator: HARA Toshiro 九州大学, 医学(系)研究科(研究院), 教授 (40150445)
• Co-Investigator(Kenkyū-buntansha): TAKADA Hidetoshi 九州大学, 医学研究院, 教授 (70294931) ; IHARA Kenji 大分大学, 医学部, 教授 (80294932) ; MIYAWAKI Toshio 富山大学, 医学系研究科, 教授 (10143885) ; NAKAHATA Tatsutoshi 京都大学, iPS細胞研究所, 教授 (20110744) ; NAKATSU Yoshimichi 九州大学, 医学研究院, 准教授 (00207820)
• Co-Investigator(Renkei-kenkyūsha): ISHIKAWA Fumihiko 理化学研究所, 免疫・アレルギー科学総合研究センター・ヒト疾患モデル研究グループ, グループディレクター (30403918)
• Project Period (FY): 2010-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥44,720,000 (Direct Cost: ¥34,400,000、Indirect Cost: ¥10,320,000); Fiscal Year 2014: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000); Fiscal Year 2013: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000); Fiscal Year 2012: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000); Fiscal Year 2011: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000); Fiscal Year 2010: ¥9,880,000 (Direct Cost: ¥7,600,000、Indirect Cost: ¥2,280,000)
• Keywords: 遺伝子修復治療 / 遺伝子治療 / BTK遺伝子 / CD34陽性造血幹細胞 / iPS細胞 / アデノ.AAV.ベクター

Outline of Final Research Achievements

We constructed a HD-Ad.AAV BTK targeting vector that is composed of the genomic sequence containing BTK exons 6-19, TIMM8A and GFP/hygromycin resistant gene. By using HD-Ad.AAV. BTK vector, we found that the targeting in the BTK gene occurred in 4 among 755 hygromycin-resistant BFU-E colonies differentiated from the cord blood CD34+ cells. Importantly, the gene targeting was also observed in CD19+ lymphoid progenitor cells that could be differentiated from the homologous recombinants of CD34+ cells. Our study shows the potential of HD-Ad.AAV. BTK vector for the BTK gene therapy by homologous recombination in hematopoietic stem cells.

Research Products

• [Journal Article] Successful treatment of non-Hodgkin's lymphoma using R-CHOP in a patient with Wiskott-Aldrich syndrome followed by a reduced-intensity stem cell transplant. (2014)
  • Author(s): Yuhki Koga, Hidetoshi Takada, Aiko Suminoe, Shouichi Ohga, Toshiro Hara
  • Journal Title: Pediatric Transplantation Volume: VOL.18 Issue: 6
  • DOI: 10.1111/petr.12297
  • Peer Reviewed / Open Access
• [Journal Article] 限られたウイルスに易感染性を示す免疫不全症（単純ヘルペス脳炎、EBV、パピローマウイルス、細胞融解型感染形式をとるウイルス） (2014)
  • Author(s): 石村匡崇、高田英俊、原 寿郎
  • Journal Title: 小児内科 Volume: 46巻 Pages: 1470-1474
• [Journal Article] B cell function after unrelated umbilical cord blood transplantation using minimal-intensity conditioning regimen in patients with X-SCID. (2013)
  • Author(s): Kumaki S, Sasahara Y, Kamachi Y, Muramatsu H, Morio T, Goi K, Sugita K, Urabe T, Takada H, Kojima S, Tsuchiya S, Hara T
  • Journal Title: Int J Hematol Volume: 98 Issue: 3 Pages: 355-60
  • DOI: 10.1007/s12185-013-1408-7
  • Peer Reviewed
• [Journal Article] A non-invasive diagnosis of histiocytic necrotizing lymphadenitis by means of gene expression profile analysis of peripheral blood mononuclear cells (2013)
  • Author(s): Ishimura M, Yamamoto H, Mizuno Y, Takada H, Goto M, Doi T, Hoshina T, Ohga S, Ohshima K, Hara T
  • Journal Title: J Clin Immunol Volume: 33(5) Issue: 5 Pages: 1018-26
  • DOI: 10.1007/s10875-013-9897-y
  • Peer Reviewed
• [Journal Article] 2.幹細胞異常と内科系疾患、現状と展望 1）造血幹細胞の異常：先天性免疫不全症 (2013)
  • Author(s): 原 寿郎
  • Journal Title: 日本内科学雑誌 Volume: 102(9) Pages: 2255-61
• [Journal Article] NKRP1A+ γδ and αβ T cells are preferentially induced in patients with Salmonella infection. (2012)
  • Author(s): Hoshina T
  • Journal Title: Hum Immunol Volume: 73 Issue: 4 Pages: 623-628
  • DOI: 10.1111/j.1365-2265.2012.04390.x
  • Peer Reviewed
• [Journal Article] Clinical and host genetic characteristics of Mendelian susceptibility to mycobacterial diseases in Japan (2011)
  • Author(s): Hoshina T, Takada H, Sasaki-Mihara Y, Kusuhara K, Ohshima K, Okada S, Kobayashi M, Ohara O, Hara T
  • Journal Title: J Clin Immunol Volume: 31 Pages: 309-314
  • Peer Reviewed
• [Journal Article] Potent induction of IFN-Y production from cord blood NK cells by the stimulation with single-stranded RNA (2011)
  • Author(s): Eljaafari FM, Takada H, Tanaka T, Doi T, Ohga S, Hara T
  • Journal Title: J Clin Immunol Volume: 31 Pages: 728-735
  • Peer Reviewed
• [Journal Article] Nationwide Survey of Patients with Primary Immunodeficiency Diseases in Japan (2011)
  • Author(s): Ishimura M, Takada H, Doi T, Imai K, Sasahara Y, Kanegane H, Nishikomori R, Morio T, Heike T, Kobayashi M, Ariga T, Tsuchiya S, Nonoyama S, Miyawaki T, Hara T
  • Journal Title: J Clin Immunol Volume: 31 Pages: 968-976
  • NAID: 120005133089
  • Peer Reviewed
• [Journal Article] PD1 as a common candidate susceptibility gene of subacute sclerosing panencephalitis. (2010)
  • Author(s): Ishizaki Y, et al.
  • Journal Title: Hum Genet Volume: 127 Pages: 411-419
  • Peer Reviewed
• [Journal Article] Smad2 and Smad3 are redundantly essential for the TGF-β-mediated regulation of regulatory T plasticity and Th1 development. (2010)
  • Author(s): Takimoto T, et al.
  • Journal Title: J Immunol Volume: 185 Pages: 842-55
  • Peer Reviewed
• [Journal Article] Clinical Features and Outcome of Patients With IRAK-4 and My D88 Deficiency. (2010)
  • Author(s): Picard C, et al.
  • Journal Title: Medicine(Baltimore) Volume: 89 Pages: 403-425
  • Peer Reviewed
• [Presentation] アデノ・アデノ随伴ウイルスベクターを用いた相同組換えによるBTK遺伝子修復 (2014)
  • Author(s): 石村匡崇、山元裕之、落合正行、高田英俊、土居岳彦、楠原浩一、中津可道、續輝久、三谷幸之介、原寿郎
  • Organizer: 第117回日本小児科学会学術集会
  • Place of Presentation: 名古屋国際会議場（愛知県名古屋市）
  • Year and Date: 2014-04-11 – 2014-04-13
• [Presentation] Autoimmune diseases in patients with primary immunodeficiency diseases. (2013)
  • Author(s): Hara T, Ishimura M, Tahada H
  • Organizer: The 2nd International Congress on Controversies in Rheumatology and Autoimmunity
  • Place of Presentation: Budapest, Hungary
• [Presentation] Innate immunity and infection in the newborn (2013)
  • Author(s): Hara T
  • Organizer: The 9th Congress of Asian Society for Pediatric Research
  • Place of Presentation: Kuching, Sarawak, Malaysia
  • Invited
• [Presentation] A critical role of the innate immune system to host defense in infants. (2012)
  • Author(s): Hara T
  • Organizer: The 8th Congress of Asian Society for Pediatric Research
  • Place of Presentation: Seoul, Korea
  • Invited
• [Presentation] Pediatric Research From Bedside to Bench to Bedside (2011)
  • Author(s): Hara T.
  • Organizer: PAS/ASPR
  • Place of Presentation: Denver, U.S.A.(招待講演)
• [Presentation] A Critical Role of Innate Immunity Receptors in the Development of Kawasaki Disease (2010)
  • Author(s): Hara T.
  • Organizer: Pediatric Academic Societies.
  • Place of Presentation: Vancouver, USA
• [Presentation] BTK gene repair by homologous recombination using helper-dependent adenovirus/adeno-associated virus hybrid vector
  • Author(s): 山元裕之
  • Organizer: 日本免疫学会
  • Place of Presentation: 幕張メッセ（千葉県）
• [Book] 希少疾患／難病の診断・治療と製品開発 (2013)
  • Author(s): 原 寿郎
  • Total Pages: 17
  • Publisher: （株）技術情報協会
• [Book] 免疫学コア講義 改訂3版 (2012)
  • Author(s): 原 寿郎
  • Publisher: 南山堂
• [Book] 今日の治療と看護(改訂第3版)原発性免疫不全症候群 (2012)
  • Author(s): 原寿郎(分担)
  • Publisher: 医学書院(印刷中)
• [Book] 今日の診断指針 症候編第6版 自己炎症性症候群 (2010)
  • Author(s): 原寿郎
  • Publisher: 医学書院(印刷中)