Here we examined the effect of EtOH on the pluripotency of undifferentiated hESCs as a model of early FAE that includes the pre-implantation period. Specifically, we demonstrated that a 24 hr low dose (20 mM) EtOH treatment significantly reduced the pluripotency (differentiation potential) of hESCs. While to our knowledge no other hESC studies have addressed this issue, somewhat analogous observations were made with rhesus monkey ESCs, albeit at much higher EtOH concentrations and over a course of 4 weeks (VandeVoort et al., 2011). We also, for the first time, demonstrated overall increases in DNA methylation in hESCs, defined the genetic and epigenetic molecular landscapes affected by low dose EtOH exposure, and identified genome-wide hotspots that could potentially be vulnerable to FAE. Furthermore, we have identified landscapes of molecular networks that are potentially deregulated by EtOH exposure through DNA methylomic alterations. This is in contrast to the recently reported lack of methylation changes after exposure of hESCs to 20 mM EtOH (Krishnamoorthy et al., 2013). Interestingly, their reported selective gene subsets are, for the most part, also unchanged in our study,
