Orthogonal Strategies for Profiling Potential Cellular Targets of Anandamide and Cannabidiol

The human endocannabinoid system regulates a myriad of physiological processes through a complex lipid signaling network involving cannabinoids and their respective receptors, cannabinoid receptor 1 (hCB1R) and cannabinoid receptor 2 (hCB2R). Anandamide (AEA) and cannabidiol (CBD) are classical examples of cannabinoids that elicit a variety of effects, both beneficial and detrimental, through these receptors. Mounting evidence suggested the presence of other potential cannabinoid targets that may be responsible for other observable effects. However, prior pharmacological studies on these cannabinoid compounds provided scant evidence of direct engagement to these proposed targets. Moreover, to the best of our knowledge, no chemoproteomic studies have been demonstrated on CBD. Here we showed that, by taking advantage of a recently developed ‘label‐free’ 2D‐TPP (2 Dimensional‐Thermal Protein Profiling) approach, we have identified several new putative targets of both AEA and CBD. Comparison of these interaction landscapes with those obtained from well‐established affinity‐based protein profiling (AfBPP) platforms has led to the discovery of both shared and unique protein targets. Subsequent target validation of selected proteins led us to conclude that this 2D‐TPP strategy complements well with AfBPP. Affinity‐based protein profiling (AfBPP) and ‘label‐free’ 2D‐TPP (2 Dimensional‐Thermal Protein Profiling) platforms have been used to identify potential cellular targets of cannabinoids, especially cannabidiol (CBD), leading to the discovery of both shared and unique protein targets. Subsequent target validation led to the conclusion that this 2D‐TPP strategy complements well with AfBPP.