Authors

Discussion

Kinetoplastid parasites continue to pose a burden on global health and current treatments for these infections are largely toxic and difficult to administer. With drug resistant strains emerging, it is important to identify new targets for therapeutics. Flap endonucleases are essential enzymes involved in DNA replication and repair. The critical role of flap endonucleases in these processes has led to interest in the human enzyme as a potential target in cancer therapy. In the current study we focus on investigating the potential of the T. brucei flap endonuclease as a novel therapeutic target. We over-expressed both a native and catalytically inert mutant form of the parasite enzyme in T. brucei blood stream form cells, using a tetracycline-inducible system. The catalytically inert protein had a severely detrimental effect on cell growth, and morphological changes were observed 72 hours post-induction. We also targeted the recombinant enzyme with a commercially available flavonoid, which is known to inhibit the human homologue. This resulted in inhibition of enzymatic activity in vitro, and critically was shown to be effective at killing parasites. This study highlights the identification of a novel therapeutic target in kinetoplastids. Further work is in progress to identify selective inhibitors of the T. brucei enzyme and to determine on-target effects in the parasite.