Developing the Universal iPS cell lines using genetic engineering system

• Project/Area Number: 26290066
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Medical genome science
• Research Institution: Tokai University
• Principal Investigator: KIMURA Minoru 東海大学, 医学部, 教授 (10146706)
• Co-Investigator(Kenkyū-buntansha): 佐藤 健人 東海大学, 医学部, 准教授 (50235363)
• Research Collaborator: SHIINA Takashi 東海大学, 医学部, 准教授 (00317744)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,900,000 (Direct Cost: ¥13,000,000、Indirect Cost: ¥3,900,000); Fiscal Year 2016: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2015: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
• Keywords: iPS細胞 / 移植片拒絶回避 / HLA遺伝子 / 遺伝子欠失 / CRISPR/Cas9 / 遺伝子発現制御 / NK細胞 / HLA発現低下 / ベータ２m遺伝子 / 移植片拒絶 / HLA / CRISPR / 遺伝子発現 / 再生 / 遺伝子工学 / HLA抗原 / 発現抑制 / 遺伝子変異 / 遺伝子改変 / MHC / 主要組織適合抗原 / 多分化能

Outline of Final Research Achievements

When stocked iPSCs are utilized to allograft transplantation, immune responses to transplanted cells should be controlled. Since construction of stocked iPSCs covering all HLA allo-types are beyond the realm of possibility, various approaches to overcome the difficulty are to be explored. While we easily manipulated HLA gene locus by CRISPR/Cas9 system, it was suggested that some selection cassettes harboring PB sequences is useful to select genome-edited cells. It was raised the possibility that beta;2m-deificient iPSCs would be the target of NK cells. Therefore, instead of all HLA class I, deletion of single HLA gene locus, such as HLA-A, would be effective to apply stocked iPSCs to patients who share the HLA allo-type other than the deleted HLA-A gene locus. It is also important to establish the in vivo experimental system in which human immune cells are re-constructed and the reactions to iPSC-derived allo antigens are validated.

Research Products

• [Journal Article] Reduced immunocompetent B cells and increased secondary infection in elderly patients with severe sepsis. (2016)
  • Author(s): K.Suzuki, S.Inoue, Y.Kametani, Y.Komori, S.Chiba, T.Sato, S.Inokuchi, S.Ogura
  • Journal Title: Shock Volume: 46 Pages: 270-278
  • Peer Reviewed / Open Access
• [Journal Article] Identification of a novel HLA-C allele, HLA-C*03:313, in a Japanese individual. HLA(Tissue Antigen) (2016)
  • Author(s): S. Suzuki T. Sato, A. Akatsuka1, M. Kimura1 and T. Stiina
  • Journal Title: HLA (Tissue Antigen) Volume: 87 Pages: 186-187
  • DOI: 10.1111/tan.12750
  • Peer Reviewed
• [Journal Article] Identification of a novel HLA-C allele, HLA-C*03:313, in a Japanese Indivisual (2016)
  • Author(s): Suzuki, S., Sato, T., Akatsuka, H., Kimura, M. and Shiina, T.
  • Journal Title: HLA Volume: 87 Pages: 186-187
  • Peer Reviewed / Open Access
• [Presentation] Ambra1はT細胞レセプターに下流で制御されるミトコンドリアの生合成と品質管理に重要である (2016)
  • Author(s): 久我周平、赤塚尚子、大塚正人、木村穣、井ノ上逸朗、佐藤健人
  • Organizer: 第３９回日本分子生物学会年会
  • Place of Presentation: パシフフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-11-30
• [Presentation] シアル酸修飾制御はリソソーム内分解のほかオートファジー開始にも重要である (2016)
  • Author(s): 赤塚尚子、Odontuya Davaadorj、木村穣、鈴木崇弘、関島良樹、大野田秀樹、佐藤健人
  • Organizer: 第３９回日本分子生物学会年会
  • Place of Presentation: パシフフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-11-30
• [Presentation] コーデン症候群患者由来iPS細胞の樹立と解析 (2015)
  • Author(s): 赤塚尚子、大塚正人、木村穣、井ノ上逸朗、佐藤健人
  • Organizer: 第38回日本分子生物学会年会・第88回日本生化学会大会合同大会
  • Place of Presentation: 兵庫県神戸市、神戸ポートアイランド
  • Year and Date: 2015-12-01