In microglia, we identified four distinct states that did not adhere closely to the classical M1/M2 (inflammatory/anti-inflammatory) distinction35. Indeed, recent studies are beginning to question the biological accuracy of the widely-cited M1/M2 classification59. An increased proportion of microglia showed an inflammatory gene expression profile in those with AUD. Chronic alcohol exposure has been shown to cause microglial activation in mice, leading to neuroinflammation60. In microglia, we identified a sole regulatory module driven by ZBTB16, a negative regulator of inflammation61, that exhibited reduced expression in individuals with AUD. Therefore, this decrease in ZBTB16-mediated regulation may further exacerbate inflammatory responses in microglia in AUD.
