In addition to identifying EtOH-induced global changes in genetic and epigenetic molecular landscapes, we were able to narrow down these changes further to four major potential signaling pathways with help of extensive bioinformatics analyses. To our knowledge, this is the first time that energy metabolism, neuroactive ligand-receptor interaction, vascular smooth muscle contraction, and calcium signaling pathways are implicated in EtOH-induced changes in human embryonic stem cells. We provided strong molecular and bioinformatic evidence that even at a low physiologically relevant dose, EtOH may epigenetically alter key genes such as SLC12A4, P2RX3 and others in glycerophospholipid metabolism and regulation of calcium ion transport and/or calcium-mediated signaling, which ultimately may lead to the significant decrease in self-renewal capacity of hESCs.
