Profiling G protein-coupled receptors of Fasciola hepatica identifies orphan rhodopsins unique to phylum Platyhelminthes

Abstract

AbstractG protein-coupled receptors (GPCRs) are established drug targets. Despite their considerable appeal as targets for next-generation anthelmintics, poor understanding of their diversity and function in parasitic helminths has thwarted progress towards GPCR-targeted anti-parasite drugs. This study facilitates GPCR research in the liver fluke, Fasciola hepatica, by generating the first profile of GPCRs from the F. hepatica genome. Our dataset describes 146 high confidence GPCRs, representing the largest cohort of GPCRs, and the most complete set of in silico ligand-receptor predictions, yet reported in any parasitic helminth. All GPCRs fall within the established GRAFS nomenclature; comprising three glutamate, 135 rhodopsin, two adhesion, five frizzled and one smoothened GPCR. Stringent annotation pipelines identified 18 highly diverged rhodopsins in F. hepatica that maintained core rhodopsin signatures, but lacked significant similarity with non-flatworm sequences, providing a new sub-group of potential flukicide targets. These facilitated identification of a larger cohort of 76 related sequences from available flatworm genomes, representing new members of existing groups of flatworm-specific rhodopsins. These receptors imply flatworm specific GPCR functions, and/or co-evolution with unique flatworm ligands, and could facilitate development of exquisitely selective anthelminthics. Ligand binding domain sequence conservation relative to deorphanised rhodopsins enabled high confidence ligand-receptor matching of seventeen receptors activated by acetylcholine, neuropeptide F/Y, octopamine or serotonin. RNA-Seq analyses showed expression of 101 GPCRs across various developmental stages, with the majority expressed most highly in the pathogenic intra-mammalian juvenile parasites. These data identify a broad complement of GPCRs in F. hepatica, including rhodopsins likely to have key functions in neuromuscular control and sensory perception, as well as frizzled and adhesion families implicated, in other species, in growth, development and reproduction. This catalogue of liver fluke GPCRs provides a platform for new avenues into our understanding of flatworm biology and anthelmintic discovery.Author SummaryFasciola spp. liver fluke are important veterinary pathogens with impacts on human and animal health, and food security, around the world. Liver fluke have developed resistance to most of the drugs used to treat them (flukicides). Since no vaccines exist, we need to develop new flukicides as a matter of urgency. Most anthelmintic drugs used to treat parasitic worm infections operate by impeding the functioning of their nerve and muscle. In flatworms, most nervous signals are received by a type of receptor called a G protein-coupled receptor (GPCR). Since GPCRs control important parasite functions (e.g. movement, egg-laying, feeding), they represent appealing targets for new flukicides, but have not yet been targeted as such. This work exploited the F. hepatica genome to determine the quantity and diversity of GPCRs in liver fluke. We found more GPCRs in the Fasciola genome than have been reported in any other parasitic worm. These findings provide a foundation that for researchers to determine the functions of these receptors, and which molecules/ligands they are activated by. These data will pave the way to exploring the potential of F. hepatica GPCRs as targets for new flukicides.