Genetic analysis of praziquantel resistance in schistosome parasites implicates a Transient Receptor Potential channel

10 Jun 2021

Mass treatment with praziquantel (PZQ) monotherapy is the mainstay for schistosome treatment. This drug shows imperfect cure rates in the field and parasites showing reduced response to PZQ can be selected in the laboratory, but the extent of resistance in Schistosoma mansoni populations is unknown. We examined the genetic basis of variation in PZQ response in a S. mansoni population (SmLE-PZQ-R) selected with PZQ in the laboratory: 35% of these worms survive high dose (73 µg/mL) PZQ treatment. We used genome wide association to map loci underlying PZQ response. The major chr. 3 peak shows recessive inheritance and contains a transient receptor potential (Sm.TRPMPZQ) channel (Smp_246790), activated by nanomoles of PZQ. Marker-assisted selection of parasites at a single Sm.TRPMPZQ SNP enriched populations of PZQ-R and PZQ-S parasites showing >377 fold difference in PZQ response. The PZQ-R parasites survived treatment in rodents better than PZQ-S. Resistant parasites show 2.25-fold lower expression of Sm.TRPMPZQ than sensitive parasites. Specific chemical blockers of Sm.TRPMPZQ enhanced PZQ resistance, while Sm.TRPMPZQ activators increased sensitivity. A single SNP in Sm.TRPMPZQ differentiated PZQ-ER and PZQ-ES lines, but mutagenesis showed this was not involved in PZQ-R, suggesting linked regulatory changes. We surveyed Sm.TRPMPZQ sequence variation in 259 individual parasites from the New and Old World revealing one nonsense mutation, that results in a truncated protein with no PZQ binding site. Our results demonstrate that Sm.TRPMPZQ underlies variation in PZQ response in S. mansoni and provides an approach for monitoring emerging PZQ-resistance alleles in schistosome elimination programs.