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 Dang; David 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

doi:10.1016/j.stem.2020.07.020

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 journal › Article › Academic › peer-review

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.

Original language	English
Pages (from-to)	840-851.e6
Journal	Cell stem cell
Volume	27
Issue number	5
DOIs	https://doi.org/10.1016/j.stem.2020.07.020
Publication status	Published - 5 Nov 2020

Keywords

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

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10.1016/j.stem.2020.07.020

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Miao, Y., Ha, A., de Lau, W., Yuki, K., Santos, A. J. M., You, C., Geurts, M. H., Puschhof, J., Pleguezuelos-Manzano, C., Peng, W. C., Senlice, R., Piani, C., Buikema, J. W., Gbenedio, O. M., Vallon, M., Yuan, J., de Haan, S., Hemrika, W., Rösch, K., ... Garcia, K. C. (2020). Next-Generation Surrogate Wnts Support Organoid Growth and Deconvolute Frizzled Pleiotropy In Vivo. Cell stem cell, 27(5), 840-851.e6. https://doi.org/10.1016/j.stem.2020.07.020

@article{da881c31a6254337815abd31b6c371a3,

title = "Next-Generation Surrogate Wnts Support Organoid Growth and Deconvolute Frizzled Pleiotropy In Vivo",

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",

author = "Yi Miao and Andrew Ha and {de Lau}, Wim and Kanako Yuki and Santos, {Ant{\'o}nio J M} and Changjiang You and Geurts, {Maarten H} and Jens Puschhof and Cayetano Pleguezuelos-Manzano and Peng, {Weng Chuan} and Ramazan Senlice and Carol Piani and Buikema, {Jan W} and Gbenedio, {Oghenekevwe M} and Mario Vallon and Jenny Yuan and {de Haan}, Sanne and Wieger Hemrika and Kathrin R{\"o}sch and Dang, {Luke T} and David Baker and Melanie Ott and Philippe Depeille and Wu, {Sean M} and Jarno Drost and Roeland Nusse and Roose, {Jeroen P} and Jacob Piehler and Boj, {Sylvia F} and Janda, {Claudia Y} and Hans Clevers and Kuo, {Calvin J} and Garcia, {K Christopher}",

note = "Funding Information: We acknowledge Georg Busslinger and Else Driehuis for establishing and providing stomach and pancreas organoid cultures, respectively. This work is supported by the Ludwig Foundation (K.C.G. and C.J.K.); the Mathers Fund (K.C.G.); the Howard Hughes Medical Institute (HHMI) (K.C.G. and R.N.); NIH grants U01DK085527 , U19AI116484 , R01NS100904 , and U01CA217851 (C.J.K.); and NIH grant 1R01DK115728 (C.J.K. and K.C.G.). P.D. is a Mark Foundation Momentum Fellow supported by the Mark Foundation for Cancer Research . Publisher Copyright: {\textcopyright} 2020 Elsevier Inc. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

year = "2020",

month = nov,

day = "5",

doi = "10.1016/j.stem.2020.07.020",

language = "English",

volume = "27",

pages = "840--851.e6",

journal = "Cell stem cell",

issn = "1934-5909",

publisher = "Cell Press",

number = "5",

}

Miao, Y, Ha, A, de Lau, W, Yuki, K, Santos, AJM, You, C, Geurts, MH, Puschhof, J, Pleguezuelos-Manzano, C, Peng, WC, Senlice, R, Piani, C, Buikema, JW, Gbenedio, OM, Vallon, M, Yuan, J, de Haan, S, Hemrika, W, Rösch, K, Dang, LT, Baker, D, Ott, M, Depeille, P, Wu, SM, Drost, J, Nusse, R, Roose, JP, Piehler, J, Boj, SF, Janda, CY, Clevers, H, Kuo, CJ & Garcia, KC 2020, 'Next-Generation Surrogate Wnts Support Organoid Growth and Deconvolute Frizzled Pleiotropy In Vivo', Cell stem cell, vol. 27, no. 5, pp. 840-851.e6. https://doi.org/10.1016/j.stem.2020.07.020

TY - JOUR

T1 - Next-Generation Surrogate Wnts Support Organoid Growth and Deconvolute Frizzled Pleiotropy In Vivo

AU - Miao, Yi

AU - Ha, Andrew

AU - de Lau, Wim

AU - Yuki, Kanako

AU - Santos, António J M

AU - You, Changjiang

AU - Geurts, Maarten H

AU - Puschhof, Jens

AU - Pleguezuelos-Manzano, Cayetano

AU - Peng, Weng Chuan

AU - Senlice, Ramazan

AU - Piani, Carol

AU - Buikema, Jan W

AU - Gbenedio, Oghenekevwe M

AU - Vallon, Mario

AU - Yuan, Jenny

AU - de Haan, Sanne

AU - Hemrika, Wieger

AU - Rösch, Kathrin

AU - Dang, Luke T

AU - Baker, David

AU - Ott, Melanie

AU - Depeille, Philippe

AU - Wu, Sean M

AU - Drost, Jarno

AU - Nusse, Roeland

AU - Roose, Jeroen P

AU - Piehler, Jacob

AU - Boj, Sylvia F

AU - Janda, Claudia Y

AU - Clevers, Hans

AU - Kuo, Calvin J

AU - Garcia, K Christopher

N1 - Funding Information: We acknowledge Georg Busslinger and Else Driehuis for establishing and providing stomach and pancreas organoid cultures, respectively. This work is supported by the Ludwig Foundation (K.C.G. and C.J.K.); the Mathers Fund (K.C.G.); the Howard Hughes Medical Institute (HHMI) (K.C.G. and R.N.); NIH grants U01DK085527 , U19AI116484 , R01NS100904 , and U01CA217851 (C.J.K.); and NIH grant 1R01DK115728 (C.J.K. and K.C.G.). P.D. is a Mark Foundation Momentum Fellow supported by the Mark Foundation for Cancer Research . Publisher Copyright: © 2020 Elsevier Inc. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.

PY - 2020/11/5

Y1 - 2020/11/5

N2 - 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.

AB - 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.

KW - DARPin

KW - Frizzled

KW - Wnt

KW - canonical Wnt signaling

KW - organoids

KW - protein engineering

KW - regenerative medicine

KW - stem cell

KW - surrogate Wnt

UR - http://www.scopus.com/inward/record.url?scp=85090222968&partnerID=8YFLogxK

U2 - 10.1016/j.stem.2020.07.020

DO - 10.1016/j.stem.2020.07.020

M3 - Article

C2 - 32818433

SN - 1934-5909

VL - 27

SP - 840-851.e6

JO - Cell stem cell

JF - Cell stem cell

IS - 5

ER -

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

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Keywords

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