analysis of postmortem brain tissues cannot determine whether the dysregulation of brain miRNAs and mRNAs was due to pre-existing vulnerability factors (such as genetic variants and/or environmental insults) or long-term alcohol consumption. We intended to verify AUD-associated brain miRNA and mRNA changes using ethanol-exposed hESC-derived cortical interneurons as models. However, not many AUD-associated brain miRNAs and mRNAs were validated by the in vitro cellular model study. This is likely due to cell-type differences between brain tissues (containing many different types of neurons and glial cells) and hESC-derived neural cultures (mainly cortical interneurons and a small number of glial cells). To confirm whether AUD-associated brain miRNA and mRNA expression changes were indeed due to alcohol use and occur in a certain type of brain neuronal or glial cells, we could use controlled animal model studies and single-cell (or nucleus) RNA-seq.
