2. Academic Validation
  3. Next-Generation Surrogate Wnts Support Organoid Growth and Deconvolute Frizzled Pleiotropy In Vivo

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

  • Cell Stem Cell. 2020 Nov 5;27(5):840-851.e6. doi: 10.1016/j.stem.2020.07.020.
Yi Miao 1,  Andrew Ha 2,  Wim de Lau 3,  Kanako Yuki 4,  António J M Santos 4,  Changjiang You 5,  Maarten H Geurts 3,  Jens Puschhof 3,  Cayetano Pleguezuelos-Manzano 3,  Weng Chuan Peng 6,  Ramazan Senlice 7,  Carol Piani 7,  Jan W Buikema 8,  Oghenekevwe M Gbenedio 9,  Mario Vallon 4,  Jenny Yuan 4,  Sanne de Haan 10,  Wieger Hemrika 11,  Kathrin Rösch 12,  Luke T Dang 13,  David Baker 13,  Melanie Ott 12,  Philippe Depeille 8,  Sean M Wu 14,  Jarno Drost 10,  Roeland Nusse 15,  Jeroen P Roose 9,  Jacob Piehler 5,  Sylvia F Boj 7,  Claudia Y Janda 16,  Hans Clevers 17,  Calvin J Kuo 4,  K Christopher Garcia 18
Affiliations

Affiliations

  • 1 Department of Molecular and Cellular Physiology, Department of Structural Biology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 2 Department of Medicine, Division of Hematology, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 3 Oncode Institute, Hubrecht Institute, University Medical Centre Utrecht, Utrecht, the Netherlands.
  • 4 Department of Medicine, Division of Hematology, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 5 Division of Biophysics, Department of Biology, University of Osnabrück, 49076 Osnabrück, Germany.
  • 6 Howard Hughes Medical Institute, Department of Developmental Biology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA; Princess Máxima Center for Pediatric Oncology, Utrecht, the Netherlands.
  • 7 Foundation Hubrecht Organoid Technology (HUB), Utrecht, the Netherlands.
  • 8 Department of Cardiology, University Medical Center Utrecht & Utrecht Regenerative Medicine Center, Utrecht University, 3508 GA Utrecht, the Netherlands.
  • 9 Department of Anatomy, University of California, San Francisco, San Francisco, CA, USA.
  • 10 Oncode Institute, Princess Máxima Center for Pediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, the Netherlands.
  • 11 U-Protein Express BV, Yalelaan 62, 3584 CM Utrecht, the Netherlands.
  • 12 Gladstone Institutes and Department of Medicine, University of California, San Francisco, San Francisco, CA, USA.
  • 13 Department of Biochemistry, Institute for Protein Design and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98105, USA.
  • 14 Division of Cardiovascular Medicine, Department of Medicine, Cardiovascular Institute and Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 15 Howard Hughes Medical Institute, Department of Developmental Biology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 16 Princess Máxima Center for Pediatric Oncology, Utrecht, the Netherlands.
  • 17 Oncode Institute, Hubrecht Institute, University Medical Centre Utrecht, Utrecht, the Netherlands; Oncode Institute, Princess Máxima Center for Pediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, the Netherlands.
  • 18 Department of Molecular and Cellular Physiology, Department of Structural Biology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA 94305, USA. Electronic address: [email protected].
PMID: 32818433 DOI: 10.1016/j.stem.2020.07.020
Abstract

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.

Keywords

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

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