Abstract text

Introduction: Fasciolosis is a globally prevalent zoonosis and a neglected tropical disease caused by infection with liver flukes such as Fasciola hepatica. Due to resistance against the main drug in use, triclabendazole, new drug candidates are urgently needed. Protein kinases (PKs) might serve as starting point for the development of new drugs, because they regulate a vast variety of cellular processes including cell growth, proliferation, differentiation and metabolism.

Objective: We pursue the hypothesis that inhibition of PKs may represent a promising therapeutic strategy against fasciolosis. The aim of this project is to identify druggable kinases in F. hepatica and potent PK inhibitors.

Material & Methods: To generate a draft kinome dataset of F. hepatica we employ a genomic-bioinformatic approach using the programs Kinnanote, OrthoMCL, Exonerate, pBLAT and InterProScan to generate a kinome dataset of F.hepatica. This dataset will allow the prioritization of potential target kinases followed by virtual screening of compound libraries against homology models of these prioritized kinases. The top hit compounds will be screened against different life stages of F. hepatica in vitro, and their inhibitory activity confirmed against recombinantly expressed PKs. Genetic target validation will be achieved by RNA interference (RNAi) against selected kinases.

Results: A first F. hepatica draft kinome contains 225 PKs to form 9 (sub) families. Single-cell transcriptomics data, show that some prioritized PKs are predominately expressed in cells and tissues of therapeutic interest, such as neoblasts, the tegument and neuronal cells. PK inhibitor treatment had lethal activity against both, immature and adult worms 24 hours after treatment at concentrations comparable to triclabendazole.

Conclusion: Preliminary data suggest that targeting PKs may represent an effective approach to control F. hepatica. Further investigations will assess the re-purposing potential of PK inhibitors against this parasitic flatworm.