Ros3 (Lem3p/CDC50) gene dosage is implicated in miltefosine susceptibility in Leishmania (Viannia) braziliensis clinical isolates and in Leishmania (Leishmania) major

Espada, C. R., Albuquerque-Wendt, A., Hornillos, V., Gluenz, E. , Coelho, A. C. and Uliana, S. R.B. (2021) Ros3 (Lem3p/CDC50) gene dosage is implicated in miltefosine susceptibility in Leishmania (Viannia) braziliensis clinical isolates and in Leishmania (Leishmania) major. ACS Infectious Diseases, 7(4), pp. 849-858. (doi: 10.1021/acsinfecdis.0c00857) (PMID:33724800) (PMCID:PMC8042657)

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Abstract

The Ros3 protein is a component of the MT-Ros3 transporter complex, considered as the main route of miltefosine entry in Leishmania. L. braziliensis clinical isolates presenting differences in miltefosine susceptibility and uptake were previously shown to differentially express ros3. In this work, we showed that the ros3 gene copy number was increased in the isolate presenting the highest rates of miltefosine uptake and, thus, the highest susceptibility to this drug. The role of the ros3 gene dosage in miltefosine susceptibility was then investigated through a modulation of the gene copy number using two distinct approaches: through an overexpression of ros3 in a tolerant L. braziliensis clinical isolate and in L. major and by generating mono- and diallelic knockouts of this gene in L. major using clustered regularly interspaced short palindromic repeats (CRISPR) Cas9 (Cas = CRISPR-associated). Although the levels of ros3 mRNA were increased at least 40-fold in overexpressing clones, no significant reduction in the half-maximal effective concentration (EC50) for miltefosine was observed in these parasites. The partial or complete deletion of ros3 in L. major, in turn, resulted in a significant increase of 3 and 20 times, respectively, in the EC50 to miltefosine. We unequivocally showed that the ros3 copy number is one of the factors involved in the differential susceptibility and uptake of miltefosine.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Gluenz, Dr Eva and Albuquerque-Wendt, Ms Andreia
Authors: Espada, C. R., Albuquerque-Wendt, A., Hornillos, V., Gluenz, E., Coelho, A. C., and Uliana, S. R.B.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:ACS Infectious Diseases
Publisher:American Chemical Society
ISSN:2373-8227
ISSN (Online):2373-8227
Published Online:16 March 2021
Copyright Holders:Copyright © 2021 American Chemical Society
First Published:First published in ACS Infectious Diseases 7(4): 849-858
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
170547The Wellcome Centre for Molecular Parasitology ( Core Support )Andrew WatersWellcome Trust (WELLCOTR)104111/Z/14/ZRIII - Parasitology