Abrogation of E-Cadherin-Mediated Cell–Cell Contact in Mouse Embryonic Stem Cells Results in Reversible LIF-Independent Self-Renewal

Abstract

Abstract We have previously demonstrated that differentiation of embryonic stem (ES) cells is associated with downregulation of cell surface E-cadherin. In this study, we assessed the function of E-cadherin in mouse ES cell pluripotency and differentiation. We show that inhibition of E-cadherin-mediated cell–cell contact in ES cells using gene knockout (Ecad−/−), RNA interference (EcadRNAi), or a transhomodimerization-inhibiting peptide (CHAVC) results in cellular proliferation and maintenance of an undifferentiated phenotype in fetal bovine serum-supplemented medium in the absence of leukemia inhibitory factor (LIF). Re-expression of E-cadherin in Ecad−/−, EcadRNAi, and CHAVC-treated ES cells restores cellular dependence to LIF supplementation. Although reversal of the LIF-independent phenotype in Ecad−/− ES cells is dependent on the β-catenin binding domain of E-cadherin, we show that β-catenin null (βcat−/−) ES cells also remain undifferentiated in the absence of LIF. This suggests that LIF-independent self-renewal of Ecad−/− ES cells is unlikely to be via β-catenin signaling. Exposure of Ecad−/−, EcadRNAi, and CHAVC-treated ES cells to the activin receptor-like kinase inhibitor SB431542 led to differentiation of the cells, which could be prevented by re-expression of E-cadherin. To confirm the role of transforming growth factor β family signaling in the self-renewal of Ecad−/− ES cells, we show that these cells maintain an undifferentiated phenotype when cultured in serum-free medium supplemented with Activin A and Nodal, with fibroblast growth factor 2 required for cellular proliferation. We conclude that transhomodimerization of E-cadherin protein is required for LIF-dependent ES cell self-renewal and that multiple self-renewal signaling networks subsist in ES cells, with activity dependent upon the cellular context. Disclosure of potential conflicts of interest is found at the end of this article.