GPR35 inhibits TRPA1-mediated colonic afferent hypersensitivity through suppression of Substance P release

The development of non-opioid analgesics for the treatment of chronic abdominal pain is a pressing area of unmet clinical need. To address this, we examined the expression of Gi/o-coupled receptors in colonic sensory neurons, which, like opioid receptors, have the potential to inhibit nociceptor activation due to their inhibitory G protein coupling. This led to the identification of the orphan receptor GPR35 as a visceral analgesic drug target due to its marked co-expression with TRPA1, a mediator of noxious mechanotransduction in the bowel. Consistent with in silico docking studies which identified binding sites for the mast cell stabiliser cromolyn and phosphodiesterase inhibitor zaprinast at GPR35, we demonstrated, using GPR35 knockout mice, that the antinociceptive effects of these drugs on TRPA1-mediated colonic nociceptor activation and mechanosensitisation were GPR35-dependent. Further work showed these antinociceptive effects occurred through the inhibition of substance P (SP) release. This confirmed both the pronociceptive effect of SP on colonic afferents, and the contribution of SP to TRPA1-mediated colonic nociceptor activation and sensitisation. We also found that TRPA1-induced contraction of the colon was mediated by SP signalling and could be inhibited by cromolyn in a GPR35-dependent manner. Our data identify GPR35, through its inhibition of SP-mediated colonic contractility and nociceptor activation and sensitisation, as a putative mechanism for the reported clinical efficacy of cromolyn in the treatment of irritable bowel syndrome. These findings highlight the potential utility of GPR35 agonists to deliver non-opioid analgesia for the treatment of abdominal pain associated with gastrointestinal diseases such as irritable bowel syndrome and inflammatory bowel disease. ### Competing Interest Statement A.J.H.B. and R.S. are employed by Sosei-Heptares. D.C.B. receives research funding from Sosei-Heptares.