Abstract
Alcohol use contributes to numerous diseases and injuries. The nervous system is affected by alcohol in diverse ways, though the molecular mechanisms of these effects are not clearly understood. Using human-induced pluripotent stem cells (iPSCs), we developed a neural cell culture model to identify the mechanisms of alcohol’s effects. iPSCs were generated from fibroblasts and differentiated into forebrain neural cells cultures that were treated with 50 mM alcohol or sham conditions (same media lacking alcohol) for 7 days. We analyzed gene expression using total RNA sequencing (RNA-seq) for 34 samples derived from 10 subjects and for 10 samples from 5 subjects in an independent experiment that had intermittent exposure to the same dose of alcohol. We also analyzed genetic effects on gene expression and conducted a weighted correlation network analysis. We found that differentiated neural cell cultures have the capacity to recapitulate gene regulatory effects previously observed in specific primary neural tissues and identified 226 genes that were differentially expressed (FDR < 0.1) after alcohol treatment. The effects on expression included decreases in INSIG1 and LDLR, two genes involved in cholesterol homeostasis. We also identified a module of 58 co-expressed genes that were uniformly decreased following alcohol exposure. The majority of these effects were supported in independent alcohol exposure experiments. Enrichment analysis linked the alcohol responsive genes to cell cycle, notch signaling, and cholesterol biosynthesis pathways, which are disrupted in several neurological disorders. Our findings suggest that there is convergence between these disorders and the effects of alcohol exposure.
Highlights
Alcohol is commonly consumed worldwide[1]
Human-induced pluripotent stem cells Fibroblasts obtained from subjects enrolled in studies at the University of Connecticut Health Center (Farmington, CT)[21,22,23] were reprogrammed to iPSCs using CytoTune retro- or sendai virus kits (Thermo Fisher Scientific) by the University of Connecticut Stem Cell Core (Farmington, CT) and cultured on irradiated mouse embryonic fibroblasts as we have previously described in detail[24,25,26]
SNPs that were previously identified as eQTLs in cortex tissue had effects that ranked significantly higher among the eQTLs identified in iPSC-derived neural cultures (anterior cingulate cortex (BA24), p = 0.001; cortex, p = 0.003; and frontal cortex (BA9)), p = 0.012; Fig. 2)
Summary
Moderate intake of alcohol may have modest health benefits[2,3], its misuse significantly contributes to numerous diseases and injuries from accidents[1,4]. Alcohol consumption can progress to the development of an alcohol use disorder (AUD). AUD affects nearly 14% of the U.S population and is characterized by tolerance to alcohol’s effects, continued use despite adverse consequences, and the development of withdrawal symptoms upon reducing alcohol intake[5,6]. It remains to be fully elucidated how the structural changes in the brain may relate to the development and progression of AUD. Our understanding of the effects of alcohol at the molecular level in human neural cells is limited, due in part to the challenges associated with obtaining and Jensen et al Translational Psychiatry (2019)9:96
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