Herein, the presented results reveal the complex mechanism of CBD action in OA. We have identified the most relevant molecular targets of CBD and the spinal cord as the main site of its action. Our results highlight the importance of molecular changes regarding pain pathway sensitization occurring in OA as a confounding factor in the analgesic efficacy of drugs. We identified the PPAR? receptor as the most influential target for beneficial CBD effects in OA. On the other hand, we uncovered that TRP channel upregulation in OA and activation by CBD is a significant disruptive factor for CBD analgesic effects (see Figure 11 for the schematic representation of key factors in the CBD mechanism of action). Our results pave the way for novel CBD-based therapy for OA with improved therapeutic potential but also encourage the use of bioinformatic tools to predict the CBD mechanism of action in different conditions.