RNA-seq of CBD-treated SK-N-BE(2) cells over a time course of 3hr - 24hr with vehicle (ethanol) matched controls at each time point.

Cannabidiol (CBD) is an FDA approved drug for treatment of drug-intractable forms of pediatric epilepsy. While a unified mechanism of action has yet to emerge from numerous proposed molecular targets of CBD, preclinical studies reveal a pleiotropic pharmacology. Here, we report the application of temporally-resolved multi-omic profiling and biosensor screening to dissect the molecular mechanism of CBD in human neuroblastoma cells. CDB treatment led to a rise in cytosolic calcium and activation of AMPK signaling within 2 hours. Subcellular profiling of proteins, metabolites, and mRNA transcripts identified CBD-dependent activation of cholesterol biosynthesis, transport and storage. We found that CBD incorporates into cellular membranes, alters cholesterol activity, and increases lipid order. We also report CBD-induced apoptosis in multiple human cell lines, which is rescued by statins and potentiated by compounds that disrupt cholesterol trafficking and storage. Our data point to pharmacological interaction of CBD with cholesterol homeostasis pathways, with potential implications in its therapeutic use. mRNA profiles of CBD or vehicle (ethanol, 0.1% v/v)-treated SK-N-BE(2) cells. Four time points (3h, 6h, 12h, and 24h) were profiled at 20µM, with four replicates of CBD treatment and four replicates vehicle treatment at each time point (total of 32 samples).