Chemical genomic analysis of GPR35 signalingElectronic supplementary information (ESI) available: shRNA hit identification, cAMP and Ca2+ flux assay results, kinase shRNA hits, gene expression data, and other assay data. See DOI: 10.1039/c7ib00005g

GPR35, a family A orphan G protein-coupled receptor, has been implicated in inflammatory, neurological, and cardiovascular diseases. However, not much is known about the signaling and functions of GPR35. We performed a label-free kinome short hairpin RNA screen and identified a putative signaling network of GPR35 in HT-29 cells, some of which was validated using gene expression, biochemical and cellular assays. The results showed that GPR35 induced hypoxia-inducible factor 1α, and was involved in synaptic transmission, sensory perception, the immune system, and morphogenetic processes. Collectively, our data suggest that GPR35 may play an important role in response to hypoxic stress and be a potential target for the treatment of inflammatory, cardiovascular, and neurological disorders. Label-free human kinome shRNA screening and DNA microarray gene expression analysis offer distinct, but complementary, views of GPR35 signaling.