Functional and genetic evidence that nucleoside transport is highly conserved in Leishmania species: Implications for pyrimidine-based chemotherapy

Alzahrani, K. J.H., Ali, J. A.M., Eze, A. A., Looi, W. L., Tagoe, D. N.A., Creek, D. J., Barrett, M. P. and De Koning, H. P. (2017) Functional and genetic evidence that nucleoside transport is highly conserved in Leishmania species: Implications for pyrimidine-based chemotherapy. International Journal for Parasitology: Drugs and Drug Resistance, 7(2), pp. 206-226. (doi:10.1016/j.ijpddr.2017.04.003) (PMID:28453984) (PMCID:PMC5407577)

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Abstract

Leishmania pyrimidine salvage is replete with opportunities for therapeutic intervention with enzyme inhibitors or antimetabolites. Their uptake into cells depends upon specific transporters; therefore it is essential to establish whether various Leishmania species possess similar pyrimidine transporters capable of drug uptake. Here, we report a comprehensive characterization of pyrimidine transport in L. major and L. mexicana. In both species, two transporters for uridine/adenosine were detected, one of which also transported uracil and the antimetabolites 5-fluoruracil (5-FU) and 5F,2′deoxyuridine (5F,2′dUrd), and was designated uridine-uracil transporter 1 (UUT1); the other transporter mediated uptake of adenosine, uridine, 5F,2′dUrd and thymidine and was designated Nucleoside Transporter 1 (NT1). To verify the reported L. donovani model of two NT1-like genes encoding uridine/adenosine transporters, and an NT2 gene encoding an inosine transporter, we cloned the corresponding L. major and L. mexicana genes, expressing each in T. brucei. Consistent with the L. donovani reports, the NT1-like genes of either species mediated the adenosine-sensitive uptake of [3H]-uridine but not of [3H]-inosine. Conversely, the NT2-like genes mediated uptake of [3H]-inosine but not [3H]-uridine. Among pyrimidine antimetabolites tested, 5-FU and 5F,2′dUrd were the most effective antileishmanials; resistance to both analogs was induced in L. major and L. mexicana. In each case it was found that the resistant cells had lost the transport capacity for the inducing drug. Metabolomics analysis found that the mechanism of action of 5-FU and 5F-2′dUrd was similar in both Leishmania species, with major changes in deoxynucleotide metabolism. We conclude that the pyrimidine salvage system is highly conserved in Leishmania species - essential information for the development of pyrimidine-based chemotherapy.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Barrett, Professor Michael and Creek, Dr Darren and Alzahrani, Khalid Jamaan and De Koning, Professor Harry
Authors: Alzahrani, K. J.H., Ali, J. A.M., Eze, A. A., Looi, W. L., Tagoe, D. N.A., Creek, D. J., Barrett, M. P., and De Koning, H. P.
College/School:College of Medical Veterinary and Life Sciences
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:International Journal for Parasitology: Drugs and Drug Resistance
Publisher:Elsevier
ISSN:2211-3207
ISSN (Online):2211-3207
Published Online:20 April 2017
Copyright Holders:Copyright © 2017 The Authors
First Published:First published in International Journal for Parasitology: Drugs and Drug Resistance 7(2):206-226
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
590521Wellcome 096984/Z - D TagoeHarry De KoningWellcome Trust (WELLCOTR)096984/Z/11/ZIII - PARASITOLOGY
590522Wellcome 096984/Z - D TagoeHarry De KoningWellcome Trust (WELLCOTR)096984/Z/11/ZIII - PARASITOLOGY
371796The Wellcome Centre for Molecular Parasitology ( Core Support )Andrew WatersWellcome Trust (WELLCOTR)085349/Z/08/ZIII - PARASITOLOGY
371798The Wellcome Centre for Molecular Parasitology ( Core Support )Andrew WatersWellcome Trust (WELLCOTR)085349/B/08/ZIII - PARASITOLOGY