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Anti-Sclerostin Antibody Inhibits Internalization of Sclerostin and Sclerostin-Mediated Antagonism of Wnt/LRP6 Signaling

Sclerosteosis is a rare high bone mass disease that is caused by inactivating mutations in the SOST gene. Its gene product, Sclerostin, is a key negative regulator of bone formation and might therefore serve as a target for the anabolic treatment of osteoporosis. The exact molecular mechanism by whi...

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Autores principales: van Dinther, Maarten, Zhang, Juan, Weidauer, Stella E., Boschert, Verena, Muth, Eva-Maria, Knappik, Achim, de Gorter, David J. J., van Kasteren, Puck B., Frisch, Christian, Mueller, Thomas D., ten Dijke, Peter
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3639248/
https://www.ncbi.nlm.nih.gov/pubmed/23638027
http://dx.doi.org/10.1371/journal.pone.0062295
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author van Dinther, Maarten
Zhang, Juan
Weidauer, Stella E.
Boschert, Verena
Muth, Eva-Maria
Knappik, Achim
de Gorter, David J. J.
van Kasteren, Puck B.
Frisch, Christian
Mueller, Thomas D.
ten Dijke, Peter
author_facet van Dinther, Maarten
Zhang, Juan
Weidauer, Stella E.
Boschert, Verena
Muth, Eva-Maria
Knappik, Achim
de Gorter, David J. J.
van Kasteren, Puck B.
Frisch, Christian
Mueller, Thomas D.
ten Dijke, Peter
author_sort van Dinther, Maarten
collection PubMed
description Sclerosteosis is a rare high bone mass disease that is caused by inactivating mutations in the SOST gene. Its gene product, Sclerostin, is a key negative regulator of bone formation and might therefore serve as a target for the anabolic treatment of osteoporosis. The exact molecular mechanism by which Sclerostin exerts its antagonistic effects on Wnt signaling in bone forming osteoblasts remains unclear. Here we show that Wnt3a-induced transcriptional responses and induction of alkaline phosphatase activity, an early marker of osteoblast differentiation, require the Wnt co-receptors LRP5 and LRP6. Unlike Dickkopf1 (DKK1), Sclerostin does not inhibit Wnt-3a-induced phosphorylation of LRP5 at serine 1503 or LRP6 at serine 1490. Affinity labeling of cell surface proteins with [(125)I]Sclerostin identified LRP6 as the main specific Sclerostin receptor in multiple mesenchymal cell lines. When cells were challenged with Sclerostin fused to recombinant green fluorescent protein (GFP) this was internalized, likely via a Clathrin-dependent process, and subsequently degraded in a temperature and proteasome-dependent manner. Ectopic expression of LRP6 greatly enhanced binding and cellular uptake of Sclerostin-GFP, which was reduced by the addition of an excess of non-GFP-fused Sclerostin. Finally, an anti-Sclerostin antibody inhibited the internalization of Sclerostin-GFP and binding of Sclerostin to LRP6. Moreover, this antibody attenuated the antagonistic activity of Sclerostin on canonical Wnt-induced responses.
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spelling pubmed-36392482013-05-01 Anti-Sclerostin Antibody Inhibits Internalization of Sclerostin and Sclerostin-Mediated Antagonism of Wnt/LRP6 Signaling van Dinther, Maarten Zhang, Juan Weidauer, Stella E. Boschert, Verena Muth, Eva-Maria Knappik, Achim de Gorter, David J. J. van Kasteren, Puck B. Frisch, Christian Mueller, Thomas D. ten Dijke, Peter PLoS One Research Article Sclerosteosis is a rare high bone mass disease that is caused by inactivating mutations in the SOST gene. Its gene product, Sclerostin, is a key negative regulator of bone formation and might therefore serve as a target for the anabolic treatment of osteoporosis. The exact molecular mechanism by which Sclerostin exerts its antagonistic effects on Wnt signaling in bone forming osteoblasts remains unclear. Here we show that Wnt3a-induced transcriptional responses and induction of alkaline phosphatase activity, an early marker of osteoblast differentiation, require the Wnt co-receptors LRP5 and LRP6. Unlike Dickkopf1 (DKK1), Sclerostin does not inhibit Wnt-3a-induced phosphorylation of LRP5 at serine 1503 or LRP6 at serine 1490. Affinity labeling of cell surface proteins with [(125)I]Sclerostin identified LRP6 as the main specific Sclerostin receptor in multiple mesenchymal cell lines. When cells were challenged with Sclerostin fused to recombinant green fluorescent protein (GFP) this was internalized, likely via a Clathrin-dependent process, and subsequently degraded in a temperature and proteasome-dependent manner. Ectopic expression of LRP6 greatly enhanced binding and cellular uptake of Sclerostin-GFP, which was reduced by the addition of an excess of non-GFP-fused Sclerostin. Finally, an anti-Sclerostin antibody inhibited the internalization of Sclerostin-GFP and binding of Sclerostin to LRP6. Moreover, this antibody attenuated the antagonistic activity of Sclerostin on canonical Wnt-induced responses. Public Library of Science 2013-04-29 /pmc/articles/PMC3639248/ /pubmed/23638027 http://dx.doi.org/10.1371/journal.pone.0062295 Text en © 2013 van Dinther et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
van Dinther, Maarten
Zhang, Juan
Weidauer, Stella E.
Boschert, Verena
Muth, Eva-Maria
Knappik, Achim
de Gorter, David J. J.
van Kasteren, Puck B.
Frisch, Christian
Mueller, Thomas D.
ten Dijke, Peter
Anti-Sclerostin Antibody Inhibits Internalization of Sclerostin and Sclerostin-Mediated Antagonism of Wnt/LRP6 Signaling
title Anti-Sclerostin Antibody Inhibits Internalization of Sclerostin and Sclerostin-Mediated Antagonism of Wnt/LRP6 Signaling
title_full Anti-Sclerostin Antibody Inhibits Internalization of Sclerostin and Sclerostin-Mediated Antagonism of Wnt/LRP6 Signaling
title_fullStr Anti-Sclerostin Antibody Inhibits Internalization of Sclerostin and Sclerostin-Mediated Antagonism of Wnt/LRP6 Signaling
title_full_unstemmed Anti-Sclerostin Antibody Inhibits Internalization of Sclerostin and Sclerostin-Mediated Antagonism of Wnt/LRP6 Signaling
title_short Anti-Sclerostin Antibody Inhibits Internalization of Sclerostin and Sclerostin-Mediated Antagonism of Wnt/LRP6 Signaling
title_sort anti-sclerostin antibody inhibits internalization of sclerostin and sclerostin-mediated antagonism of wnt/lrp6 signaling
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3639248/
https://www.ncbi.nlm.nih.gov/pubmed/23638027
http://dx.doi.org/10.1371/journal.pone.0062295
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