Non-equivalence of Wnt and R-spondin ligands during Lgr5 + intestinal stem-cell self-renewal

Kelley S. Yan, Claudia Y. Janda, Junlei Chang, Grace X.Y. Zheng, Kathryn A. Larkin, Vincent C. Luca, Luis A. Chia, Amanda T. Mah, Arnold Han, Jessica M. Terry, Akifumi Ootani, Kelly Roelf, Mark Lee, Jenny Yuan, Xiao Li, Christopher R. Bolen, Julie Wilhelmy, Paige Davies, Hiroo Ueno, Richard J. Von FurstenbergPhillip Belgrader, Solongo B. Ziraldo, Heather Ordonez, Susan J. Henning, Melissa Wong, Michael P. Snyder, Irving L. Weissman, Aaron J. Hsueh, Tarjei S. Mikkelsen, K. Christopher Garcia, Calvin J. Kuo

Research output: Contribution to journalArticle

97 Citations (Scopus)

Abstract

The canonical Wnt/β-catenin signalling pathway governs diverse developmental, homeostatic and pathological processes. Palmitoylated Wnt ligands engage cell-surface frizzled (FZD) receptors and LRP5 and LRP6 co-receptors, enabling β-catenin nuclear translocation and TCF/LEF-dependent gene transactivation. Mutations in Wnt downstream signalling components have revealed diverse functions thought to be carried out by Wnt ligands themselves. However, redundancy between the 19 mammalian Wnt proteins and 10 FZD receptors and Wnt hydrophobicity have made it difficult to attribute these functions directly to Wnt ligands. For example, individual mutations in Wnt ligands have not revealed homeostatic phenotypes in the intestinal epithelium-an archetypal canonical, Wnt pathway-dependent, rapidly self-renewing tissue, the regeneration of which is fueled by proliferative crypt Lgr5 + intestinal stem cells (ISCs). R-spondin ligands (RSPO1-RSPO4) engage distinct LGR4-LGR6, RNF43 and ZNRF3 receptor classes, markedly potentiate canonical Wnt/β-catenin signalling, and induce intestinal organoid growth in vitro and Lgr5 + ISCs in vivo. However, the interchangeability, functional cooperation and relative contributions of Wnt versus RSPO ligands to in vivo canonical Wnt signalling and ISC biology remain unknown. Here we identify the functional roles of Wnt and RSPO ligands in the intestinal crypt stem-cell niche. We show that the default fate of Lgr5 + ISCs is to differentiate, unless both RSPO and Wnt ligands are present. However, gain-of-function studies using RSPO ligands and a new non-lipidated Wnt analogue reveal that these ligands have qualitatively distinct, non-interchangeable roles in ISCs. Wnt proteins are unable to induce Lgr5 + ISC self-renewal, but instead confer a basal competency by maintaining RSPO receptor expression that enables RSPO ligands to actively drive and specify the extent of stem-cell expansion. This functionally non-equivalent yet cooperative interaction between Wnt and RSPO ligands establishes a molecular precedent for regulation of mammalian stem cells by distinct priming and self-renewal factors, with broad implications for precise control of tissue regeneration.

Original languageEnglish (US)
Pages (from-to)238-242
Number of pages5
JournalNature
Volume545
Issue number7653
DOIs
StatePublished - May 11 2017

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Ligands
Stem Cells
Catenins
Wnt Proteins
Frizzled Receptors
Wnt Signaling Pathway
Cell Self Renewal
Regeneration
Organoids
Stem Cell Niche
Mutation
Cell Surface Receptors
Pathologic Processes
Intestinal Mucosa
Hydrophobic and Hydrophilic Interactions
Transcriptional Activation
Cell Biology
Phenotype
Growth
Genes

ASJC Scopus subject areas

  • Medicine(all)
  • General

Cite this

Yan, K. S., Janda, C. Y., Chang, J., Zheng, G. X. Y., Larkin, K. A., Luca, V. C., ... Kuo, C. J. (2017). Non-equivalence of Wnt and R-spondin ligands during Lgr5 + intestinal stem-cell self-renewal. Nature, 545(7653), 238-242. https://doi.org/10.1038/nature22313

Non-equivalence of Wnt and R-spondin ligands during Lgr5 + intestinal stem-cell self-renewal. / Yan, Kelley S.; Janda, Claudia Y.; Chang, Junlei; Zheng, Grace X.Y.; Larkin, Kathryn A.; Luca, Vincent C.; Chia, Luis A.; Mah, Amanda T.; Han, Arnold; Terry, Jessica M.; Ootani, Akifumi; Roelf, Kelly; Lee, Mark; Yuan, Jenny; Li, Xiao; Bolen, Christopher R.; Wilhelmy, Julie; Davies, Paige; Ueno, Hiroo; Von Furstenberg, Richard J.; Belgrader, Phillip; Ziraldo, Solongo B.; Ordonez, Heather; Henning, Susan J.; Wong, Melissa; Snyder, Michael P.; Weissman, Irving L.; Hsueh, Aaron J.; Mikkelsen, Tarjei S.; Garcia, K. Christopher; Kuo, Calvin J.

In: Nature, Vol. 545, No. 7653, 11.05.2017, p. 238-242.

Research output: Contribution to journalArticle

Yan, KS, Janda, CY, Chang, J, Zheng, GXY, Larkin, KA, Luca, VC, Chia, LA, Mah, AT, Han, A, Terry, JM, Ootani, A, Roelf, K, Lee, M, Yuan, J, Li, X, Bolen, CR, Wilhelmy, J, Davies, P, Ueno, H, Von Furstenberg, RJ, Belgrader, P, Ziraldo, SB, Ordonez, H, Henning, SJ, Wong, M, Snyder, MP, Weissman, IL, Hsueh, AJ, Mikkelsen, TS, Garcia, KC & Kuo, CJ 2017, 'Non-equivalence of Wnt and R-spondin ligands during Lgr5 + intestinal stem-cell self-renewal', Nature, vol. 545, no. 7653, pp. 238-242. https://doi.org/10.1038/nature22313
Yan KS, Janda CY, Chang J, Zheng GXY, Larkin KA, Luca VC et al. Non-equivalence of Wnt and R-spondin ligands during Lgr5 + intestinal stem-cell self-renewal. Nature. 2017 May 11;545(7653):238-242. https://doi.org/10.1038/nature22313
Yan, Kelley S. ; Janda, Claudia Y. ; Chang, Junlei ; Zheng, Grace X.Y. ; Larkin, Kathryn A. ; Luca, Vincent C. ; Chia, Luis A. ; Mah, Amanda T. ; Han, Arnold ; Terry, Jessica M. ; Ootani, Akifumi ; Roelf, Kelly ; Lee, Mark ; Yuan, Jenny ; Li, Xiao ; Bolen, Christopher R. ; Wilhelmy, Julie ; Davies, Paige ; Ueno, Hiroo ; Von Furstenberg, Richard J. ; Belgrader, Phillip ; Ziraldo, Solongo B. ; Ordonez, Heather ; Henning, Susan J. ; Wong, Melissa ; Snyder, Michael P. ; Weissman, Irving L. ; Hsueh, Aaron J. ; Mikkelsen, Tarjei S. ; Garcia, K. Christopher ; Kuo, Calvin J. / Non-equivalence of Wnt and R-spondin ligands during Lgr5 + intestinal stem-cell self-renewal. In: Nature. 2017 ; Vol. 545, No. 7653. pp. 238-242.
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abstract = "The canonical Wnt/β-catenin signalling pathway governs diverse developmental, homeostatic and pathological processes. Palmitoylated Wnt ligands engage cell-surface frizzled (FZD) receptors and LRP5 and LRP6 co-receptors, enabling β-catenin nuclear translocation and TCF/LEF-dependent gene transactivation. Mutations in Wnt downstream signalling components have revealed diverse functions thought to be carried out by Wnt ligands themselves. However, redundancy between the 19 mammalian Wnt proteins and 10 FZD receptors and Wnt hydrophobicity have made it difficult to attribute these functions directly to Wnt ligands. For example, individual mutations in Wnt ligands have not revealed homeostatic phenotypes in the intestinal epithelium-an archetypal canonical, Wnt pathway-dependent, rapidly self-renewing tissue, the regeneration of which is fueled by proliferative crypt Lgr5 + intestinal stem cells (ISCs). R-spondin ligands (RSPO1-RSPO4) engage distinct LGR4-LGR6, RNF43 and ZNRF3 receptor classes, markedly potentiate canonical Wnt/β-catenin signalling, and induce intestinal organoid growth in vitro and Lgr5 + ISCs in vivo. However, the interchangeability, functional cooperation and relative contributions of Wnt versus RSPO ligands to in vivo canonical Wnt signalling and ISC biology remain unknown. Here we identify the functional roles of Wnt and RSPO ligands in the intestinal crypt stem-cell niche. We show that the default fate of Lgr5 + ISCs is to differentiate, unless both RSPO and Wnt ligands are present. However, gain-of-function studies using RSPO ligands and a new non-lipidated Wnt analogue reveal that these ligands have qualitatively distinct, non-interchangeable roles in ISCs. Wnt proteins are unable to induce Lgr5 + ISC self-renewal, but instead confer a basal competency by maintaining RSPO receptor expression that enables RSPO ligands to actively drive and specify the extent of stem-cell expansion. This functionally non-equivalent yet cooperative interaction between Wnt and RSPO ligands establishes a molecular precedent for regulation of mammalian stem cells by distinct priming and self-renewal factors, with broad implications for precise control of tissue regeneration.",
author = "Yan, {Kelley S.} and Janda, {Claudia Y.} and Junlei Chang and Zheng, {Grace X.Y.} and Larkin, {Kathryn A.} and Luca, {Vincent C.} and Chia, {Luis A.} and Mah, {Amanda T.} and Arnold Han and Terry, {Jessica M.} and Akifumi Ootani and Kelly Roelf and Mark Lee and Jenny Yuan and Xiao Li and Bolen, {Christopher R.} and Julie Wilhelmy and Paige Davies and Hiroo Ueno and {Von Furstenberg}, {Richard J.} and Phillip Belgrader and Ziraldo, {Solongo B.} and Heather Ordonez and Henning, {Susan J.} and Melissa Wong and Snyder, {Michael P.} and Weissman, {Irving L.} and Hsueh, {Aaron J.} and Mikkelsen, {Tarjei S.} and Garcia, {K. Christopher} and Kuo, {Calvin J.}",
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T1 - Non-equivalence of Wnt and R-spondin ligands during Lgr5 + intestinal stem-cell self-renewal

AU - Yan, Kelley S.

AU - Janda, Claudia Y.

AU - Chang, Junlei

AU - Zheng, Grace X.Y.

AU - Larkin, Kathryn A.

AU - Luca, Vincent C.

AU - Chia, Luis A.

AU - Mah, Amanda T.

AU - Han, Arnold

AU - Terry, Jessica M.

AU - Ootani, Akifumi

AU - Roelf, Kelly

AU - Lee, Mark

AU - Yuan, Jenny

AU - Li, Xiao

AU - Bolen, Christopher R.

AU - Wilhelmy, Julie

AU - Davies, Paige

AU - Ueno, Hiroo

AU - Von Furstenberg, Richard J.

AU - Belgrader, Phillip

AU - Ziraldo, Solongo B.

AU - Ordonez, Heather

AU - Henning, Susan J.

AU - Wong, Melissa

AU - Snyder, Michael P.

AU - Weissman, Irving L.

AU - Hsueh, Aaron J.

AU - Mikkelsen, Tarjei S.

AU - Garcia, K. Christopher

AU - Kuo, Calvin J.

PY - 2017/5/11

Y1 - 2017/5/11

N2 - The canonical Wnt/β-catenin signalling pathway governs diverse developmental, homeostatic and pathological processes. Palmitoylated Wnt ligands engage cell-surface frizzled (FZD) receptors and LRP5 and LRP6 co-receptors, enabling β-catenin nuclear translocation and TCF/LEF-dependent gene transactivation. Mutations in Wnt downstream signalling components have revealed diverse functions thought to be carried out by Wnt ligands themselves. However, redundancy between the 19 mammalian Wnt proteins and 10 FZD receptors and Wnt hydrophobicity have made it difficult to attribute these functions directly to Wnt ligands. For example, individual mutations in Wnt ligands have not revealed homeostatic phenotypes in the intestinal epithelium-an archetypal canonical, Wnt pathway-dependent, rapidly self-renewing tissue, the regeneration of which is fueled by proliferative crypt Lgr5 + intestinal stem cells (ISCs). R-spondin ligands (RSPO1-RSPO4) engage distinct LGR4-LGR6, RNF43 and ZNRF3 receptor classes, markedly potentiate canonical Wnt/β-catenin signalling, and induce intestinal organoid growth in vitro and Lgr5 + ISCs in vivo. However, the interchangeability, functional cooperation and relative contributions of Wnt versus RSPO ligands to in vivo canonical Wnt signalling and ISC biology remain unknown. Here we identify the functional roles of Wnt and RSPO ligands in the intestinal crypt stem-cell niche. We show that the default fate of Lgr5 + ISCs is to differentiate, unless both RSPO and Wnt ligands are present. However, gain-of-function studies using RSPO ligands and a new non-lipidated Wnt analogue reveal that these ligands have qualitatively distinct, non-interchangeable roles in ISCs. Wnt proteins are unable to induce Lgr5 + ISC self-renewal, but instead confer a basal competency by maintaining RSPO receptor expression that enables RSPO ligands to actively drive and specify the extent of stem-cell expansion. This functionally non-equivalent yet cooperative interaction between Wnt and RSPO ligands establishes a molecular precedent for regulation of mammalian stem cells by distinct priming and self-renewal factors, with broad implications for precise control of tissue regeneration.

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