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- Title
Targeting Ergosterol Biosynthesis in Leishmania donovani: Essentiality of Sterol 14alpha-demethylase.
- Authors
McCall, Laura-Isobel; El Aroussi, Amale; Choi, Jun Yong; Vieira, Debora F.; De Muylder, Geraldine; Johnston, Jonathan B.; Chen, Steven; Kellar, Danielle; Siqueira-Neto, Jair L.; Roush, William R.; Podust, Larissa M.; McKerrow, James H.
- Abstract
Leishmania protozoan parasites (Trypanosomatidae family) are the causative agents of cutaneous, mucocutaneous and visceral leishmaniasis worldwide. While these diseases are associated with significant morbidity and mortality, there are few adequate treatments available. Sterol 14alpha-demethylase (CYP51) in the parasite sterol biosynthesis pathway has been the focus of considerable interest as a novel drug target in Leishmania. However, its essentiality in Leishmania donovani has yet to be determined. Here, we use a dual biological and pharmacological approach to demonstrate that CYP51 is indispensable in L. donovani. We show via a facilitated knockout approach that chromosomal CYP51 genes can only be knocked out in the presence of episomal complementation and that this episome cannot be lost from the parasite even under negative selection. In addition, we treated wild-type L. donovani and CYP51-deficient strains with 4-aminopyridyl-based inhibitors designed specifically for Trypanosoma cruzi CYP51. While potency was lower than in T. cruzi, these inhibitors had increased efficacy in parasites lacking a CYP51 allele compared to complemented parasites, indicating inhibition of parasite growth via a CYP51-specific mechanism and confirming essentiality of CYP51 in L. donovani. Overall, these results provide support for further development of CYP51 inhibitors for the treatment of visceral leishmaniasis. Author Summary: Visceral leishmaniasis is the second most lethal parasitic infection after malaria. Other forms of leishmaniasis also cause significant morbidity. However, there are few treatments available, and many cause severe side effects or are associated with the development of resistance. A key difference between mammalian cells and Leishmania parasites is the type of sterol in their membranes: while mammalian cell membranes contain cholesterol, Leishmania parasites use ergosterol. There has therefore been considerable interest in developing inhibitors of sterol biosynthesis pathways to target Leishmania parasites. Sterol 14alpha-demethylase (CYP51) is one of the enzymes in the sterol biosynthesis pathway, and the target of significant drug development research in Leishmania. Here we use a double approach to determine whether this gene is essential in Leishmania donovani, the causative agent of visceral leishmaniasis. We demonstrate via gene knockout and drug targeting approaches that loss or inhibition of CYP51 inhibits L. donovani growth. These results validate CYP51 as a drug target in L. donovani and support further work to develop CYP51-directed therapies for visceral leishmaniasis.
- Subjects
ERGOSTEROL; LEISHMANIA donovani; VISCERAL leishmaniasis; BIOSYNTHESIS; PARASITIC diseases; DRUG target; TRICHOMONIASIS
- Publication
PLoS Neglected Tropical Diseases, 2015, Vol 9, Issue 3, p1
- ISSN
1935-2727
- Publication type
Article
- DOI
10.1371/journal.pntd.0003588