Establishment of epidermolysis bullosa model mouse with revertant mosaicism and its treatment

• Project/Area Number: 17H01572
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Dermatology
• Research Institution: Hokkaido University
• Principal Investigator: Shimizu Hiroshi 北海道大学, 医学研究院, 教授 (00146672)
• Co-Investigator(Kenkyū-buntansha): 新熊 悟 新潟大学, 医歯学系, 准教授 (00613788) ; 岩田 浩明 北海道大学, 大学病院, 助教 (20397334) ; 西江 渉 北海道大学, 医学研究院, 准教授 (20443955) ; 乃村 俊史 北海道大学, 大学病院, 講師 (50399911) ; 藤田 靖幸 北海道大学, 医学研究院, 客員研究員 (80374437)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥43,030,000 (Direct Cost: ¥33,100,000、Indirect Cost: ¥9,930,000); Fiscal Year 2019: ¥12,350,000 (Direct Cost: ¥9,500,000、Indirect Cost: ¥2,850,000); Fiscal Year 2018: ¥12,350,000 (Direct Cost: ¥9,500,000、Indirect Cost: ¥2,850,000); Fiscal Year 2017: ¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000)
• Keywords: 復帰変異モザイク / 表皮水疱症 / 17型コラーゲン / 細胞療法 / 遺伝子編集 / 遺伝子治療 / モデルマウス / CRISPR/Cas9 / 染色体組換え / 皮膚科学

Outline of Final Research Achievements

Almost all patients with epidermolysis bullosa lacking type XVII collagen are known to have revertant mosaic spots. This revertant mosaicism is a phenomenon in which gene abnormalities are naturally repaired without any artificial modification of the gene, but the mechanism is not clear. In this study, we developed a mouse model of epidermolysis bullosa that could develop revertant mosaicism. Using this mouse, we have succeeded in inducing revertant mosaic cells, and we are now analyzing these cells.

Academic Significance and Societal Importance of the Research Achievements

近年、表皮水疱症などの遺伝性皮膚疾患で、後天的に一部の皮膚が正常化する現象（復帰変異モザイク）が報告された。そのような部位では、遺伝子異常が正常化した表皮細胞が観察されるが、この現象は尋常性魚鱗癬や先天性免疫不全などでも報告されている。現在、この復帰変異モザイク現象が生じた皮膚から採取した表皮細胞を増幅培養し自家移植する再生医療が開発されるなど、本現象は、先天性疾患に対する自家細胞を用いた根本的な再生医療の可能性をもたらす画期的な自然現象である。もしこの現象のメカニズムを解明できれば、遺伝子異常によって生じる様々な疾患の治療法の開発につながる。

Research Products

• [Journal Article] Efficient gene reframing therapy for recessive dystrophic epidermolysis bullosa using CRISPR/Cas9 (2019)
  • Author(s): Takashima Shota、Shinkuma Satoru、Fujita Yasuyuki、Nomura Toshifumi、Ujiie Hideyuki、Natsuga Ken、Iwata Hiroaki、Nakamura Hideki、Vorobyev Artem、Abe Riichiro、Shimizu Hiroshi
  • Journal Title: Journal of Investigative Dermatology Volume: in press Issue: 8 Pages: 1711-1721
  • DOI: 10.1016/j.jid.2019.02.015
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Electron Microscopic and Immunohistochemical Findings of the Epidermal Basement Membrane in Two Families with Nail-patella Syndrome (2019)
  • Author(s): Shinkuma S、Nakamura H、Maehara M、Takashima S、Nomura T、Fujita Y、Hasegawa S、Sato-Matsumura K、Abe R、Shimizu H
  • Journal Title: Acta Dermato Venereologica Volume: 99(12) Pages: 1110-1115
  • DOI: 10.2340/00015555-3318
  • Peer Reviewed / Open Access
• [Journal Article] The development of induced pluripotent stem cell-derived mesenchymal stem/stromal cells from normal human and RDEB epidermal keratinocytes (2018)
  • Author(s): Nakayama Chihiro、Fujita Yasuyuki、Matsumura Wakana、Ujiie Inkin、Takashima Shota、Shinkuma Satoru、Nomura Toshifumi、Abe Riichiro、Shimizu Hiroshi
  • Journal Title: Journal of Dermatological Science Volume: 91 Issue: 3 Pages: 301-310
  • DOI: 10.1016/j.jdermsci.2018.06.004
  • Peer Reviewed
• [Journal Article] Epidermal aspects of type VII collagen: Implications for dystrophic epidermolysis bullosa and epidermolysis bullosa acquisita (2018)
  • Author(s): Watanabe Mika、Natsuga Ken、Shinkuma Satoru、Shimizu Hiroshi
  • Journal Title: The Journal of Dermatology Volume: 45 Issue: 5 Pages: 515-521
  • DOI: 10.1111/1346-8138.14222
  • Peer Reviewed
• [Journal Article] Establishment of integration-free induced pluripotent stem cells from human recessive dystrophic epidermolysis bullosa keratinocytes (2018)
  • Author(s): Matsumura Wakana、Fujita Yasuyuki、Nakayama Chihiro、Shinkuma Satoru、Suzuki Shotaro、Nomura Toshifumi、Abe Riichiro、Shimizu Hiroshi
  • Journal Title: Journal of Dermatological Science Volume: 89 Issue: 3 Pages: 263-271
  • DOI: 10.1016/j.jdermsci.2017.11.017
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Frameshift mutations in different exons of Col17a1 lead to distinctive phenotype of junctional epidermolysis bullosa model mice. (2019)
  • Author(s): Nguyen Hong Ha, Shinkuma Satoru, Ansai Osamu, Kabata Yudai, Takashima Shota, Mori Masashi, Ikawa Masahito, Shimizu Hiroshi, Abe Riichiro
  • Organizer: The 44th Annual Meeting of the Japanese Society for Investigative Dermatology
  • Int'l Joint Research
• [Presentation] Efficient reframed gene therapy for recessive dystrophic epidermolysis bullosa using CRISPR/Cas9. (2018)
  • Author(s): Shota Takashima, Satoru Shinkuma, Yasuyuki Fujita, Toshifumi Nomura, Hideki Nakamura, Hiroshi Shimizu
  • Organizer: The 7th International Investigative Dermatology
  • Int'l Joint Research
• [Presentation] Emerging cell therapies in epidermysis bullosa. (2018)
  • Author(s): Fujita Yasuyuki
  • Organizer: The 16th International Symposium of the Cutaneous Biology Research Institute.
  • Int'l Joint Research / Invited
• [Presentation] Long lasting epithelization after cultured epidermal autograft with revertant mosaicism in recessive dystrophic epidermolysis bullosa. (2017)
  • Author(s): Matsumura W, Fujita Y, Nakamura H, Nishie W, Shinkuma S, Ujiie I, Nakayama C, Takashima S, Sawamura D, Inoie M, Shimizu H
  • Organizer: DebRA International 2017
  • Int'l Joint Research
• [Presentation] Gene-edition targeting mutation site recovers null-expression of type VII collagen caused by frameshift mutation via non-homologous end joining in recessive dystrophic epidermolysis bullosa. (2017)
  • Author(s): Takashima S, Shinkuma S, Fujita Y, Nomura T, Ujiie H, Abe R, Shimizu H
  • Organizer: The 73rd Annual Meeting of the Society for Investigative Dermatology
  • Int'l Joint Research
• [Presentation] 表皮水疱症と歩む：最近のトピックス (2017)
  • Author(s): 藤田靖幸
  • Organizer: 第116回日本皮膚科学会総会
  • Invited