Tissue tension permits β-catenin phosphorylation to drive mesoderm specification in human embryonic stem cells

Nadia M E Ayad; Johnathon N Lakins; Ajinkya Ghagre; Allen J Ehrlicher; Valerie M Weaver

doi:10.1101/2023.07.14.549074

Tissue tension permits β-catenin phosphorylation to drive mesoderm specification in human embryonic stem cells

bioRxiv [Preprint]. 2023 Jul 15:2023.07.14.549074. doi: 10.1101/2023.07.14.549074.

Authors

Nadia M E Ayad^{1

2}, Johnathon N Lakins², Ajinkya Ghagre³, Allen J Ehrlicher⁴, Valerie M Weaver^{2

5}

Affiliations

¹ Graduate Program in Bioengineering, University of California, San Francisco and University of California Berkeley, San Francisco, CA 94143, USA; Center for Bioengineering and Tissue Regeneration, Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA.
² Center for Bioengineering and Tissue Regeneration, Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA.
³ Department of Bioengineering, McGill University, Montreal, QC H3A 0E9, Canada.
⁴ Department of Bioengineering, Department of Anatomy and Cell Biology, Department of Biomedical Engineering, Department of Mechanical Engineering, Centre for Structural Biology, Rosalind and Morris Goodman Cancer Institute, McGill University, Montreal, QC H3A 1A3, Canada.
⁵ Center for Bioengineering and Tissue Regeneration, Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; UCSF Comprehensive Cancer Center, Helen Diller Family Cancer Research Center, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Bioengineering and Therapeutic Sciences, Department of Radiation Oncology, University of California, San Francisco, San Francisco, CA 94143, USA.

Abstract

The role of morphogenetic forces in cell fate specification is an area of intense interest. Our prior studies suggested that the development of high cell-cell tension in human embryonic stem cells (hESC) colonies permits the Src-mediated phosphorylation of junctional β-catenin that accelerates its release to potentiate Wnt-dependent signaling critical for initiating mesoderm specification. Using an ectopically expressed nonphosphorylatable mutant of β-catenin (Y654F), we now provide direct evidence that impeding tension-dependent Src-mediated β-catenin phosphorylation impedes the expression of Brachyury (T) and the epithelial-to-mesenchymal transition (EMT) necessary for mesoderm specification. Addition of exogenous Wnt3a or inhibiting GSK3β activity rescued mesoderm expression, emphasizing the importance of force dependent Wnt signaling in regulating mechanomorphogenesis. Our work provides a framework for understanding tension-dependent β-catenin/Wnt signaling in the self-organization of tissues during developmental processes including gastrulation.

Keywords: Wnt signaling; cytoskeletal tension; gastrulation; gastruloids; human embryonic stem cells; mechanical forces; mechanotransduction; mesoderm specification; traction force microscopy (TFM); β-catenin.

Publication types

Preprint

Abstract

Publication types

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