Simultaneous lineage tracing of multiple gastric stem cells and their paracrine regulation by stem cell niche

• Project/Area Number: 21K19474
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 53:Organ-based internal medicine and related fields
• Research Institution: The University of Tokyo
• Principal Investigator: Hayakawa Yoku 東京大学, 医学部附属病院, 講師 (60777655)
• Project Period (FY): 2021-07-09 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2022: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2021: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: 胃 / 幹細胞 / マウスモデル / 胃上皮幹細胞 / 系譜追跡 / 幹細胞ニッチ / ガストリン / R-spondin

Outline of Research at the Start

本提案では、胃上皮内幹細胞集団の動態とその制御機構について、新規のマウスモデルと単一細胞レベルの解析を用いて詳細に解析する。腺管内に存在するとされる複数の幹細胞群がどのように分裂・分化を行い、細胞間相互転換および子孫細胞の供給を行っていくのかを、多系統の細胞群を異なる蛍光蛋白で標識・追跡可能なマウスを用いて解明する。さらに、周囲傍分泌ニッチ刺激が異なる幹細胞の分裂・分化様式を誘導する様子とその機序を、単一細胞レベルの転写プロファイリングにより明らかにする。

Outline of Final Research Achievements

To examine Rspondin signaling to stem cells, we analyzed Tff1-Cre;tetO-Rspo3;R26-LSL-rtTA mice and conditional knockout mice of its receptors Lgr4 and Lgr5. As a result, Rspo3 induced differentiation into chief cells via Lgr4 expressed in stem cells and mucous neck cells. Furthermore, as a result of a lineage tracing experiment in mice in which intermediate cells located in the middle of differentiation from mucous neck cells to chief cells were selectively labeled, this intermediate cell population was divided into two fractions. The more differentiated cell type cells do not have the ability to divide, while the progenitor type cells have the function of gradual differentiation towards chief cells.

Academic Significance and Societal Importance of the Research Achievements

消化管上皮の腺管底部に存在するLgr5陽性幹細胞が同定されて以降、消化管幹細胞研究はLgr5陽性細胞の動態解析を中心に進められてきた。しかし申請者の精密かつ詳細な観察によって、胃上皮ではLgr5陽性幹細胞よりもその上方に位置するLgr5陰性（Lgr4陽性）幹細胞の方がより活発に腺管構成細胞を供給していることが明らかとなった。本研究は「これまでのLgr5=胃上皮の主たる幹細胞」という学説を覆す、大きな成果を輩出した。また、胃の基礎研究領域で簡易的な脱分化マーカーとして汎用されている主細胞・頸細胞間の中間細胞について世界で初めて選択的な細胞標識を実現し、分裂・分化様式を可視化することに成功した。

Research Products

• [Journal Article] R‐spondin signaling in the stomach: isthmal Lgr4 rules (2022)
  • Author(s): Malagola Ermanno、Hayakawa Yoku、Wang Timothy C
  • Journal Title: The EMBO Journal Volume: 41 Issue: 13
  • DOI: 10.15252/embj.2022111696
  • Peer Reviewed
• [Journal Article] R-spondin 3 governs secretory differentiation in the gastric oxyntic glands (2022)
  • Author(s): Kurokawa Ken、Wang Timothy C.、Hayakawa Yoku
  • Journal Title: Journal of Clinical Investigation Volume: 132 Issue: 21
  • DOI: 10.1172/jci163380
  • Peer Reviewed
• [Journal Article] Epithelial memory of inflammation limits tissue damage while promoting pancreatic tumorigenesis (2021)
  • Author(s): Del Poggetto Edoardo、Ho I-Lin、省略、Hayakawa Yoku、省略、Viale Andrea
  • Journal Title: Science Volume: 373 Issue: 6561 Pages: 6561-6561
  • DOI: 10.1126/science.abj0486
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Cell fate analysis of zone 3 hepatocytes in liver injury and tumorigenesis (2021)
  • Author(s): Kurosaki Shigeyuki、Nakagawa Hayato、Hayata Yuki、Kawamura Satoshi、Matsushita Yuki、Yamada Tomoharu、Uchino Koji、Hayakawa Yoku、Suzuki Nobumi、Hata Masahiro、Tsuboi Mayo、Kinoshita Hiroto、Tanaka Yasuo、Nakatsuka Takuma、Hirata Yoshihiro、Tateishi Keisuke、Koike Kazuhiko
  • Journal Title: JHEP Reports Volume: 3 Issue: 4 Pages: 100315-100315
  • DOI: 10.1016/j.jhepr.2021.100315
  • Peer Reviewed
• [Journal Article] Stem cells and origins of cancer in the upper gastrointestinal tract (2021)
  • Author(s): Hayakawa Yoku、Nakagawa Hiroshi、Rustgi Anil K.、Que Jianwen、Wang Timothy C.
  • Journal Title: Cell Stem Cell Volume: 28 Issue: 8 Pages: 1343-1361
  • DOI: 10.1016/j.stem.2021.05.012
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Helicobacter pylori CagA elicits BRCAness to induce genome instability that may underlie bacterial gastric carcinogenesis (2021)
  • Author(s): S Imai, T Ooki, N Murata-Kamiya, D Komura, K Tahmina, W Wu, Atsushi Takahashi-Kanemitsu, C T Knight, A Kunita, N Suzuki, A Del Valle, M Tsuboi, M Hata, Y Hayakawa, N Ohnishi, K Ueda, M Fukayama, T Ushiku, S Ishikawa, M Hatakeyama
  • Journal Title: Cell Host & Microbe Volume: 29 Issue: 6 Pages: 1-18
  • DOI: 10.1016/j.chom.2021.04.006
  • Peer Reviewed / Open Access / Int'l Joint Research