Genome-wide screening of the key molecules for metastasizing collagen VII-deficient squamous cell carcinoma.

• Project/Area Number: 19K08764
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53050:Dermatology-related
• Research Institution: Hokkaido University
• Principal Investigator: Nakamura Hideki 北海道大学, 医学研究院, 助手 (60435956)
• Co-Investigator(Kenkyū-buntansha): 西江 渉 北海道大学, 医学研究院, 客員教授 (20443955) ; 柳 輝希 北海道大学, 大学病院, 講師 (50755973) ; 泉 健太郎 北海道大学, 医学研究院, 助教 (50793668)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: VII型コラーゲン / 有棘細胞癌 / 遠隔転移 / 網羅的ゲノムスクリーニング / CRISPR/Cas9

Outline of Research at the Start

COL7発現をノックアウトした不死化角化細胞（HaCaT細胞）の19,500種類の遺伝子を、1細胞につき1遺伝子ずつランダムにゲノム編集し、癌化し浸潤・転移を来した細胞から標的となった遺伝子を次世代シークエンサーを用いて同定する。独創的な手法で、DEB関連SCCの浸潤・遠隔転移機構の解明を目指す

Outline of Final Research Achievements

The aim of this study was to elucidate the metastatic mechanism of squamous cell carcinoma (SCC) arising in patients with dystrophic epidermolysis bullosa (DEB) caused by mutations in type 7 collagen (COL7) gene. DEB patient model keratinocytes with a comprehensive gene-editing did not develop metastasis when transplanted subcutaneously in mice, but the subcutaneous tumors of DEB patient model showed an overall irregular structure, surrounding fibrosis, and hypervascularization compared to subcutaneous tumors of normal keratinocytes. These results suggest that subcutaneous tumors in DEB patient model induce changes in the surrounding tissue environment via abnormal construction of the basement membrane, and that these changes may contribute to the metastatic mechanism of SCC in DEB patients.

Academic Significance and Societal Importance of the Research Achievements

本研究によって、栄養障害型表皮水疱症患者（DEB）の表皮細胞は癌化する前に血管増生などの遠隔転移に繋がる組織環境を作ることが明らかとなった。今までは、線維芽細胞が主に関連するTGF-βの持続的な活性化に伴う慢性炎症がDEB患者の表皮細胞を取り囲む組織環境を変化させていると考えられてきた。本研究で得られた、DEB患者の表皮細胞自体が周囲の組織環境を遠隔転移しやすいように変化させる新たな知見は、今後DEB患者の表皮細胞を標的とした有棘細胞癌の予防法や治療法の可能性を示唆する。

Research Products

• [Journal Article] Hair follicle stem cell progeny heal blisters while pausing skin development (2021)
  • Author(s): Fujimura Yu、Watanabe Mika、Ohno Kota、Kobayashi Yasuaki、Takashima Shota、Nakamura Hideki、Kosumi Hideyuki、Wang Yunan、Mai Yosuke、Lauria Andrea、Proserpio Valentina、Ujiie Hideyuki、Iwata Hiroaki、Nishie Wataru、Nagayama Masaharu、Oliviero Salvatore、Donati Giacomo、Shimizu Hiroshi、Natsuga Ken
  • Journal Title: EMBO reports Volume: 22 Issue: 7
  • DOI: 10.15252/embr.202050882
  • NAID: 120007182923
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Intravenous allogeneic multilineage‐differentiating stress‐enduring cells in adults with dystrophic epidermolysis bullosa: a phase 1/2 open‐label study (2021)
  • Author(s): Fujita Y.、Nohara T.、Takashima S.、Natsuga K.、Adachi M.、Yoshida K.、Shinkuma S.、Takeichi T.、Nakamura H.、Wada O.、Akiyama M.、Ishiko A.、Shimizu H.
  • Journal Title: Journal of the European Academy of Dermatology and Venereology Volume: 35 Issue: 8
  • DOI: 10.1111/jdv.17201
  • Peer Reviewed
• [Journal Article] Zonula occludens-1 demonstrates a unique appearance in buccal mucosa over several layers (2021)
  • Author(s): Imafuku Keisuke、Kamaguchi Mayumi、Natsuga Ken、Nakamura Hideki、Shimizu Hiroshi、Iwata Hiroaki
  • Journal Title: Cell and Tissue Research Volume: 384 Issue: 3 Pages: 691-702
  • DOI: 10.1007/s00441-021-03425-8
  • Peer Reviewed
• [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] Cultured Epidermal Autografts from Clinically Revertant Skin as a Potential Wound Treatment for Recessive Dystrophic Epidermolysis Bullosa (2019)
  • Author(s): Matsumura Wakana、Fujita Yasuyuki、Shinkuma Satoru、Suzuki Shotaro、Yokoshiki Saki、Goto Hideki、Hayashi Hiroshi、Ono Kota、Inoie Masukazu、Takashima Shota、Nakayama Chihiro、Nomura Toshifumi、Nakamura Hideki、Abe Riichiro、Sato Norihiro、Shimizu Hiroshi
  • Journal Title: Journal of Investigative Dermatology Volume: 139(10) Issue: 10 Pages: 2115-2124.e11
  • DOI: 10.1016/j.jid.2019.03.1155
  • Peer Reviewed
• [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
• [Presentation] Wnt/β-catenin signaling regulates hemidesmosomes in keratinocytes (2021)
  • Author(s): Kosumi H, Watanabe M, Shinkuma S, Nohara T, Fujimura Y, Tsukiyama T, Donati G, Nakamura H, Iwata H, Ujiie H, Natsuga K
  • Organizer: The 46th Annual Meeting of The Japanese Society for Investigative Dermatology(Kyoto, Japan)
• [Presentation] Wnt/β-cateninシグナルが表皮角化細胞のヘミデスモソームを制御する (2021)
  • Author(s): 小住英之, 渡邉美佳, 新熊悟, 野原拓馬, 藤村悠, 築山忠維, Giacomo Donati, 岩田浩明, 中村秀樹, 氏家英之, 夏賀健.
  • Organizer: 第428回日本皮膚科学会北海道地方会（札幌、日本）
• [Presentation] 毛包幹細胞の娘細胞は皮膚の発生を停止させながら表皮下水疱を治癒させる (2021)
  • Author(s): 藤村悠, 渡邉美佳, 大野航太, 小林康明, 高島翔太, 中村秀樹, 小住英之, 王禹楠, 眞井洋輔, Andrea Lauria, Valentina Proserpio, 氏家英之, 岩田浩明, 西江渉, 長山雅晴, Salvatore Oliviero, Giacomo Donati, 清水宏, 夏賀健
  • Organizer: 第426回日本皮膚科学会北海道地方会（札幌、日本）
• [Presentation] 成人栄養障害型表皮水疱症患者を対象としたMuse細胞製品CL2020の国内1/2相試験：52週までの安全性の検討 (2021)
  • Author(s): 藤田靖幸, 野原拓馬, 高島翔太, 夏賀健, 足立太起, 吉田憲司, 新熊悟, 武市拓也, 中村秀樹, 和田理, 秋山真志, 石河晃, 清水宏.
  • Organizer: 第72回日本皮膚科学会中部支部学術大会（奈良、日本）
• [Presentation] Hair follicle stem cell progeny repair subepidermal blisters at the expense of skin development. (2020)
  • Author(s): Fujimura Y, Watanabe M, Ohno K, Kobayashi Y, Nakamura H, Lauria A, Proserpio V, Ujiie H, Iwata H, Nishie W, Nagayama M, Olivielo S, Donati G, Shimizu H, Natsuga K
  • Organizer: The 45th Annual Meeting of the Japanese Society for Investigative Dermatology
• [Presentation] Efficient gene reframing therapy for recessive dystrophic epidermolysis bullosa with CRISPR/Cas9 (2020)
  • Author(s): Takashima S, Shinkuma S, Fujita Y, Nomura T, Ujiie H, Natsuga K, Iwata H, Nakamura H, Vorobyev A, Abe R, Shimizu H
  • Organizer: The 45th Annual Meeting of the Japanese Society for Investigative Dermatology, Virtual Meeting