Proteomic Profiling Reveals Roles of Stress Response, Ca 2+ Transient Dysregulation and Novel Signaling Pathways in Alcohol‐Induced Cardiotoxicity

BACKGROUND Alcohol use in pregnancy increases the risk of abnormal cardiac development, and excessive alcohol consumption in adults can induce cardiomyopathy, contractile dysfunction, and arrhythmias. Understanding molecular mechanisms underlying alcohol-induced cardiac toxicity could provide guidance in the development of therapeutic strategies. METHODS We have performed proteomic and bioinformatic analysis to examine protein alterations globally and quantitatively in cardiomyocytes derived from human pluripotent stem cells (hiPSC-CMs) treated with ethanol. Proteins in both cell lysates and extracellular culture media were systematically quantitated. RESULTS Treatment with ethanol caused severe detrimental effects on hiPSC-CMs as indicated by significant cell death and deranged Ca2+ handling. Treatment of hiPSC-CMs with ethanol significantly affected proteins responsible for stress response (e.g. GPX1 and HSPs), ion channel related proteins (e.g. ATP1A2), myofibril structure proteins (e.g. MYL2/3), and those involved in focal adhesion and extracellular matrix (e.g. ILK and PXN). Proteins involved in the TRAF2 signaling (e.g. CPNE1 and TNIK) were also affected by ethanol treatment. CONCLUSIONS The observed changes in protein expression highlight the involvement of oxidative stress and dysregulation of Ca2+ handling and contraction while also implicating potential novel targets in alcohol-induced cardiotoxicity. These findings facilitate further exploration of potential mechanisms, discovery of novel biomarkers, and development of targeted therapeutics against ethanol-induced cardiotoxicity.