Role of protein acylation in stemness characteristics of human embryonic stem cells

Abstract

The aim of this study was to determine the effect of Porcn inhibition on human embryonic stem cells (hESCs) fate. Methods: hESCs were cultured on a feeder layer or Matrigel-coated plates. Small molecule LGK-974 was used to inhibit Porcn activity. We assessed the effect of Porcn inhibition on viability, expression of Wnt signaling targets, pluripotency markers, proliferation, differentiation and protein fatty acylation. hESCs conditioned medium (CM) containing secreted Wnt proteins was applied in rescue experiments. Results: LGK-974 at the selected concentrations showed mild effects on hESCs viability, but significantly reduced mRNA and protein expression of Wnt signaling targets (Axin-2 and c-Myc) and pluripotency markers (OCT-4 and SOX-2) (p<0.05). The Porcn inhibition showed reduced proliferation (p<0.0001), and enhanced differentiation capacity into ectodermal progenitors (SSEA-3+, NCAM+) (p<0.05), which were reverted by CM. Click chemistry reaction did not show significant alteration in protein fatty acylation upon LGK-974 treatment.