Next-Generation Surrogate Wnts Support Organoid Growth and Deconvolute Frizzled Pleiotropy In Vivo

Yi Miao, Andrew Ha, Wim de Lau, Kanako Yuki, António J M Santos, Changjiang You, Maarten H Geurts, Jens Puschhof, Cayetano Pleguezuelos-Manzano, Weng Chuan Peng, Ramazan Senlice, Carol Piani, Jan W Buikema, Oghenekevwe M Gbenedio, Mario Vallon, Jenny Yuan, Sanne de Haan, Wieger Hemrika, Kathrin Rösch, Luke T DangDavid Baker, Melanie Ott, Philippe Depeille, Sean M Wu, Jarno Drost, Roeland Nusse, Jeroen P Roose, Jacob Piehler, Sylvia F Boj, Claudia Y Janda, Hans Clevers, Calvin J Kuo, K Christopher Garcia

Research output: Contribution to journalArticlepeer-review

73 Citations (Scopus)


Modulation of Wnt signaling has untapped potential in regenerative medicine due to its essential functions in stem cell homeostasis. However, Wnt lipidation and Wnt-Frizzled (Fzd) cross-reactivity have hindered translational Wnt applications. Here, we designed and engineered water-soluble, Fzd subtype-specific "next-generation surrogate" (NGS) Wnts that hetero-dimerize Fzd and Lrp6. NGS Wnt supports long-term expansion of multiple different types of organoids, including kidney, colon, hepatocyte, ovarian, and breast. NGS Wnts are superior to Wnt3a conditioned media in organoid expansion and single-cell organoid outgrowth. Administration of Fzd subtype-specific NGS Wnt in vivo reveals that adult intestinal crypt proliferation can be promoted by agonism of Fzd5 and/or Fzd8 receptors, while a broad spectrum of Fzd receptors can induce liver zonation. Thus, NGS Wnts offer a unified organoid expansion protocol and a laboratory "tool kit" for dissecting the functions of Fzd subtypes in stem cell biology.

Original languageEnglish
Pages (from-to)840-851.e6
JournalCell stem cell
Issue number5
Publication statusPublished - 5 Nov 2020


  • DARPin
  • Frizzled
  • Wnt
  • canonical Wnt signaling
  • organoids
  • protein engineering
  • regenerative medicine
  • stem cell
  • surrogate Wnt


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