Molecular Pathway and Cell State Responsible for Dissociation-Induced Apoptosis in Human Pluripotent Stem Cells

SummaryHuman embryonic stem cells (hESCs), unlike mouse ones (mESCs), are vulnerable to apoptosis upon dissociation. Here, we show that the apoptosis, which is of a nonanoikis type, is caused by ROCK-dependent hyperactivation of actomyosin and efficiently suppressed by the myosin inhibitor Blebbistatin. The actomyosin hyperactivation is triggered by the loss of E-cadherin-dependent intercellular contact and also observed in dissociated mouse epiblast-derived pluripotent cells but not in mESCs. We reveal that Abr, a unique Rho-GEF family factor containing a functional Rac-GAP domain, is an indispensable upstream regulator of the apoptosis and ROCK/myosin hyperactivation. Rho activation coupled with Rac inhibition is induced in hESCs upon dissociation, but not in Abr-depleted hESCs or mESCs. Furthermore, artificial Rho or ROCK activation with Rac inhibition restores the vulnerability of Abr-depleted hESCs to dissociation-induced apoptosis. Thus, the Abr-dependent “Rho-high/Rac-low” state plays a decisive role in initiating the dissociation-induced actomyosin hyperactivation and apoptosis in hESCs.

► hESC dissociation causes ROCK-dependent apoptosis with extensive blebbing ► The apoptosis is caused by Rho/ROCK-mediated actomyosin hyperactivation ► Loss of E-cadherin signaling activates the Rho-GEF/Rac-GAP Abr ► The Rho-high/Rac-low state induces myosin light chain phosphorylation via ROCK