In-silico Analysis of THC/CBD-Responsive miRNA- Target Interactions and Pathway Associations in Humans

Abstract

Cannabinoids such as cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC) exert diverse immunological and cellular effects, yet the post -transcriptional mechanisms underlying these responses remain insufficiently defined . Emerging evidence suggests that microRNAs (miRNAs) may mediate cannabinoid -induced regulatory changes; however, an integrated overview of cannabinoid -responsive miRNAs and their downstream targets is lacking . This study employed an in -silico framework to systematically identify miRNAs with altered expression following THC, CBD, or combined THC +CBD exposure . Fourteen cannabinoid - responsive miRNAs were curated from human studies and animal models wit h validated human orthologs . Predicted human mRNA targets were retrieved using TargetScan, incorporating canonical seed matches and weighted context++ scores, which were subsequently classified into high-, moderate-, and low-confidence interaction groups. Target genes associated with three major signaling pathways including NF-κB/TLR, JAK/STAT, and PI3K/AKT were selected for pathway -focused analysis . A total of 3 9 miRNA-mRNA interactions were identified, with the PI3K /AKT pathway receiving the highest inter action density . Network visualization in Cytoscape revealed miR -126-5p and miR -1972 as the primary central regulators, each targeting multiple components across NF-κB/TLR, JAK/STAT, and PI3K/AKT pathways. Key genes such as STAT1, STAT3, TLR4, IRAK1, IRAK4, and AKT emerged as prominent convergence points. This study provides a mechanistic framework linking THC- and CBD-responsive miRNAs to key inflammatory, immune, and survival pathways, offering a valuable foundation for guiding future molecular validation and the development of cannabinoid-based therapeutic applications.