Aijun Zhang1,2, Lukmon Raji2, Zhiping Wu4, Sufiya Khanam1, Dehui Kong1,2, Ling Li2, Junmin Peng4, Bob Moore3, Xusheng Wang1,2, Megan Mulligan2

Cannabis is the most used federally illegal drug in the United States. Over the past years, cannabis usage has increased among adults. Growing research shows that high levels of ∆9-tetrahydrocannabinol (THC), the major psychoactive component of cannabis, impairs memory, learning, attention, cognitive performance, motivation, and is also associated with brain alterations. Accordingly, there is increasing demand to define targets of cannabinoids and to understand how individual genetic differences moderate response to THC. To address this gap, we developed a holistic approach to quantify the effects of acute THC on brain signaling by integrating multi-omics data collected from genetically divergent C57BL/6J and DBA/2J mice. Both sexes (n=4) were injected with 10 mg/kg THC or vehicle (i.p). Cortex was collected 60 mins post-injection. RNA and protein were extracted from each hemisphere, followed by RNA-seq, proteomics, and phosphoproteomics profiling. The results show that the DBA/2J strain exhibits a larger acute response to acute high dose THC than C57BL/6J, with most differences observed at the phosphosite level. Moreover, multi-omics analysis indicates that the cortical phosphoproteome responds robustly to THC at 60 mins, while whole proteome and transcriptome are less responsive. This is consistent with expectations that phosphorylation acts as an early signaling event preceding downstream transcriptional remodeling and changes in protein abundance. Our preclinical study reveals new molecular targets and signaling pathways underlying individual differences in acute responses to THC that may provide insight into genetic differences in adverse health risks in humans.

1. Department of Neurology, University of Tennessee Health Science Center, Memphis, TN 38163 USA 2. Department of Genetics, Genomics and Informatics, University of Tennessee Health Science Center, Memphis, TN 38163 USA 3. Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN 38163 USA 4. Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105 USA