Surrogate Wnt agonists that phenocopy canonical Wnt and β-catenin signalling

Claudia Y Janda; Luke T Dang; Changjiang You; Junlei Chang; Wim de Lau; Zhendong A Zhong; Kelley S Yan; Owen Marecic; Dirk Siepe; Xingnan Li; James D Moody; Bart O Williams; Hans Clevers; Jacob Piehler; David Baker; Calvin J Kuo; K Christopher Garcia

doi:10.1038/nature22306

Surrogate Wnt agonists that phenocopy canonical Wnt and β-catenin signalling

Claudia Y Janda, Luke T Dang, Changjiang You, Junlei Chang, Wim de Lau, Zhendong A Zhong, Kelley S Yan, Owen Marecic, Dirk Siepe, Xingnan Li, James D Moody, Bart O Williams, Hans Clevers, Jacob Piehler, David Baker, Calvin J Kuo, K Christopher Garcia

Hubrecht Institute

Onderzoeksoutput: Bijdrage aan wetenschappelijk tijdschrift/periodieke uitgave › Artikel › Wetenschappelijk › peer review

256 Citaten (Scopus)

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Wnt proteins modulate cell proliferation and differentiation and the self-renewal of stem cells by inducing β-catenin-dependent signalling through the Wnt receptor frizzled (FZD) and the co-receptors LRP5 and LRP6 to regulate cell fate decisions and the growth and repair of several tissues. The 19 mammalian Wnt proteins are cross-reactive with the 10 FZD receptors, and this has complicated the attribution of distinct biological functions to specific FZD and Wnt subtype interactions. Furthermore, Wnt proteins are modified post-translationally by palmitoylation, which is essential for their secretion, function and interaction with FZD receptors. As a result of their acylation, Wnt proteins are very hydrophobic and require detergents for purification, which presents major obstacles to the preparation and application of recombinant Wnt proteins. This hydrophobicity has hindered the determination of the molecular mechanisms of Wnt signalling activation and the functional importance of FZD subtypes, and the use of Wnt proteins as therapeutic agents. Here we develop surrogate Wnt agonists, water-soluble FZD-LRP5/LRP6 heterodimerizers, with FZD5/FZD8-specific and broadly FZD-reactive binding domains. Similar to WNT3A, these Wnt agonists elicit a characteristic β-catenin signalling response in a FZD-selective fashion, enhance the osteogenic lineage commitment of primary mouse and human mesenchymal stem cells, and support the growth of a broad range of primary human organoid cultures. In addition, the surrogates can be systemically expressed and exhibit Wnt activity in vivo in the mouse liver, regulating metabolic liver zonation and promoting hepatocyte proliferation, resulting in hepatomegaly. These surrogates demonstrate that canonical Wnt signalling can be activated by bi-specific ligands that induce receptor heterodimerization. Furthermore, these easily produced, non-lipidated Wnt surrogate agonists facilitate functional studies of Wnt signalling and the exploration of Wnt agonists for translational applications in regenerative medicine.

Originele taal-2	Engels
Pagina's (van-tot)	234-237
Aantal pagina's	4
Tijdschrift	Nature
Volume	545
Nummer van het tijdschrift	7653
DOI's	https://doi.org/10.1038/nature22306
Status	Gepubliceerd - 11 mei 2017

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10.1038/nature22306

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@article{c0057540d2b04d0a8337b03099710eae,

title = "Surrogate Wnt agonists that phenocopy canonical Wnt and β-catenin signalling",

abstract = "Wnt proteins modulate cell proliferation and differentiation and the self-renewal of stem cells by inducing β-catenin-dependent signalling through the Wnt receptor frizzled (FZD) and the co-receptors LRP5 and LRP6 to regulate cell fate decisions and the growth and repair of several tissues. The 19 mammalian Wnt proteins are cross-reactive with the 10 FZD receptors, and this has complicated the attribution of distinct biological functions to specific FZD and Wnt subtype interactions. Furthermore, Wnt proteins are modified post-translationally by palmitoylation, which is essential for their secretion, function and interaction with FZD receptors. As a result of their acylation, Wnt proteins are very hydrophobic and require detergents for purification, which presents major obstacles to the preparation and application of recombinant Wnt proteins. This hydrophobicity has hindered the determination of the molecular mechanisms of Wnt signalling activation and the functional importance of FZD subtypes, and the use of Wnt proteins as therapeutic agents. Here we develop surrogate Wnt agonists, water-soluble FZD-LRP5/LRP6 heterodimerizers, with FZD5/FZD8-specific and broadly FZD-reactive binding domains. Similar to WNT3A, these Wnt agonists elicit a characteristic β-catenin signalling response in a FZD-selective fashion, enhance the osteogenic lineage commitment of primary mouse and human mesenchymal stem cells, and support the growth of a broad range of primary human organoid cultures. In addition, the surrogates can be systemically expressed and exhibit Wnt activity in vivo in the mouse liver, regulating metabolic liver zonation and promoting hepatocyte proliferation, resulting in hepatomegaly. These surrogates demonstrate that canonical Wnt signalling can be activated by bi-specific ligands that induce receptor heterodimerization. Furthermore, these easily produced, non-lipidated Wnt surrogate agonists facilitate functional studies of Wnt signalling and the exploration of Wnt agonists for translational applications in regenerative medicine.",

keywords = "Animals, Cell Differentiation, Cell Lineage, Cell Proliferation, Frizzled Receptors, HEK293 Cells, Hepatocytes, Hepatomegaly, Humans, Hydrophobic and Hydrophilic Interactions, Intestines, Ligands, Liver, Low Density Lipoprotein Receptor-Related Protein-5, Low Density Lipoprotein Receptor-Related Protein-6, Mesenchymal Stromal Cells, Mice, Models, Molecular, Organoids, Protein Multimerization, Signal Transduction, Solubility, Tissue Culture Techniques, Wnt Proteins, Wnt Signaling Pathway, beta Catenin, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't",

author = "Janda, {Claudia Y} and Dang, {Luke T} and Changjiang You and Junlei Chang and {de Lau}, Wim and Zhong, {Zhendong A} and Yan, {Kelley S} and Owen Marecic and Dirk Siepe and Xingnan Li and Moody, {James D} and Williams, {Bart O} and Hans Clevers and Jacob Piehler and David Baker and Kuo, {Calvin J} and Garcia, {K Christopher}",

year = "2017",

month = may,

day = "11",

doi = "10.1038/nature22306",

language = "English",

volume = "545",

pages = "234--237",

journal = "Nature",

issn = "0028-0836",

publisher = "Nature Publishing Group",

number = "7653",

}

TY - JOUR

T1 - Surrogate Wnt agonists that phenocopy canonical Wnt and β-catenin signalling

AU - Janda, Claudia Y

AU - Dang, Luke T

AU - You, Changjiang

AU - Chang, Junlei

AU - de Lau, Wim

AU - Zhong, Zhendong A

AU - Yan, Kelley S

AU - Marecic, Owen

AU - Siepe, Dirk

AU - Li, Xingnan

AU - Moody, James D

AU - Williams, Bart O

AU - Clevers, Hans

AU - Piehler, Jacob

AU - Baker, David

AU - Kuo, Calvin J

AU - Garcia, K Christopher

PY - 2017/5/11

Y1 - 2017/5/11

N2 - Wnt proteins modulate cell proliferation and differentiation and the self-renewal of stem cells by inducing β-catenin-dependent signalling through the Wnt receptor frizzled (FZD) and the co-receptors LRP5 and LRP6 to regulate cell fate decisions and the growth and repair of several tissues. The 19 mammalian Wnt proteins are cross-reactive with the 10 FZD receptors, and this has complicated the attribution of distinct biological functions to specific FZD and Wnt subtype interactions. Furthermore, Wnt proteins are modified post-translationally by palmitoylation, which is essential for their secretion, function and interaction with FZD receptors. As a result of their acylation, Wnt proteins are very hydrophobic and require detergents for purification, which presents major obstacles to the preparation and application of recombinant Wnt proteins. This hydrophobicity has hindered the determination of the molecular mechanisms of Wnt signalling activation and the functional importance of FZD subtypes, and the use of Wnt proteins as therapeutic agents. Here we develop surrogate Wnt agonists, water-soluble FZD-LRP5/LRP6 heterodimerizers, with FZD5/FZD8-specific and broadly FZD-reactive binding domains. Similar to WNT3A, these Wnt agonists elicit a characteristic β-catenin signalling response in a FZD-selective fashion, enhance the osteogenic lineage commitment of primary mouse and human mesenchymal stem cells, and support the growth of a broad range of primary human organoid cultures. In addition, the surrogates can be systemically expressed and exhibit Wnt activity in vivo in the mouse liver, regulating metabolic liver zonation and promoting hepatocyte proliferation, resulting in hepatomegaly. These surrogates demonstrate that canonical Wnt signalling can be activated by bi-specific ligands that induce receptor heterodimerization. Furthermore, these easily produced, non-lipidated Wnt surrogate agonists facilitate functional studies of Wnt signalling and the exploration of Wnt agonists for translational applications in regenerative medicine.

AB - Wnt proteins modulate cell proliferation and differentiation and the self-renewal of stem cells by inducing β-catenin-dependent signalling through the Wnt receptor frizzled (FZD) and the co-receptors LRP5 and LRP6 to regulate cell fate decisions and the growth and repair of several tissues. The 19 mammalian Wnt proteins are cross-reactive with the 10 FZD receptors, and this has complicated the attribution of distinct biological functions to specific FZD and Wnt subtype interactions. Furthermore, Wnt proteins are modified post-translationally by palmitoylation, which is essential for their secretion, function and interaction with FZD receptors. As a result of their acylation, Wnt proteins are very hydrophobic and require detergents for purification, which presents major obstacles to the preparation and application of recombinant Wnt proteins. This hydrophobicity has hindered the determination of the molecular mechanisms of Wnt signalling activation and the functional importance of FZD subtypes, and the use of Wnt proteins as therapeutic agents. Here we develop surrogate Wnt agonists, water-soluble FZD-LRP5/LRP6 heterodimerizers, with FZD5/FZD8-specific and broadly FZD-reactive binding domains. Similar to WNT3A, these Wnt agonists elicit a characteristic β-catenin signalling response in a FZD-selective fashion, enhance the osteogenic lineage commitment of primary mouse and human mesenchymal stem cells, and support the growth of a broad range of primary human organoid cultures. In addition, the surrogates can be systemically expressed and exhibit Wnt activity in vivo in the mouse liver, regulating metabolic liver zonation and promoting hepatocyte proliferation, resulting in hepatomegaly. These surrogates demonstrate that canonical Wnt signalling can be activated by bi-specific ligands that induce receptor heterodimerization. Furthermore, these easily produced, non-lipidated Wnt surrogate agonists facilitate functional studies of Wnt signalling and the exploration of Wnt agonists for translational applications in regenerative medicine.

KW - Animals

KW - Cell Differentiation

KW - Cell Lineage

KW - Cell Proliferation

KW - Frizzled Receptors

KW - HEK293 Cells

KW - Hepatocytes

KW - Hepatomegaly

KW - Humans

KW - Hydrophobic and Hydrophilic Interactions

KW - Intestines

KW - Ligands

KW - Liver

KW - Low Density Lipoprotein Receptor-Related Protein-5

KW - Low Density Lipoprotein Receptor-Related Protein-6

KW - Mesenchymal Stromal Cells

KW - Mice

KW - Models, Molecular

KW - Organoids

KW - Protein Multimerization

KW - Signal Transduction

KW - Solubility

KW - Tissue Culture Techniques

KW - Wnt Proteins

KW - Wnt Signaling Pathway

KW - beta Catenin

KW - Journal Article

KW - Research Support, N.I.H., Extramural

KW - Research Support, Non-U.S. Gov't

U2 - 10.1038/nature22306

DO - 10.1038/nature22306

M3 - Article

C2 - 28467818

SN - 0028-0836

VL - 545

SP - 234

EP - 237

JO - Nature

JF - Nature

IS - 7653

ER -

Surrogate Wnt agonists that phenocopy canonical Wnt and β-catenin signalling

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