Functional and genetic evidence that nucleoside transport is highly conserved in Leishmania species

Implications for pyrimidine-based chemotherapy

Khalid J.H. Alzahrani, Juma A M Ali, Anthonius A. Eze, Wan Limm Looi, Daniel N.A. Tagoe, Darren J. Creek, Michael P Barrett, Harry P De Koning

Research output: Contribution to journalArticleResearchpeer-review

11 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)206-226
Number of pages21
JournalInternational Journal for Parasitology: Drugs and Drug Resistance
Volume7
Issue number2
DOIs
Publication statusPublished - 1 Aug 2017

Keywords

  • 5-fluorouracil
  • Leishmania
  • Metabolomics
  • Nucleoside transporter
  • Pyrimidine chemotherapy
  • Pyrimidine metabolism
  • Uracil transporter

Cite this

Alzahrani, Khalid J.H. ; Ali, Juma A M ; Eze, Anthonius A. ; Looi, Wan Limm ; Tagoe, Daniel N.A. ; Creek, Darren J. ; Barrett, Michael P ; De Koning, Harry P. / Functional and genetic evidence that nucleoside transport is highly conserved in Leishmania species : Implications for pyrimidine-based chemotherapy. In: International Journal for Parasitology: Drugs and Drug Resistance. 2017 ; Vol. 7, No. 2. pp. 206-226.
@article{c9ba99415fbc4835990bcb0b543c652c,
title = "Functional and genetic evidence that nucleoside transport is highly conserved in Leishmania species: Implications for pyrimidine-based chemotherapy",
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.",
keywords = "5-fluorouracil, Leishmania, Metabolomics, Nucleoside transporter, Pyrimidine chemotherapy, Pyrimidine metabolism, Uracil transporter",
author = "Alzahrani, {Khalid J.H.} and Ali, {Juma A M} and Eze, {Anthonius A.} and Looi, {Wan Limm} and Tagoe, {Daniel N.A.} and Creek, {Darren J.} and Barrett, {Michael P} and {De Koning}, {Harry P}",
year = "2017",
month = "8",
day = "1",
doi = "10.1016/j.ijpddr.2017.04.003",
language = "English",
volume = "7",
pages = "206--226",
journal = "International Journal for Parasitology: Drugs and Drug Resistance",
issn = "2211-3207",
publisher = "Elsevier",
number = "2",

}

Functional and genetic evidence that nucleoside transport is highly conserved in Leishmania species : Implications for pyrimidine-based chemotherapy. / Alzahrani, Khalid J.H.; Ali, Juma A M; Eze, Anthonius A.; Looi, Wan Limm; Tagoe, Daniel N.A.; Creek, Darren J.; Barrett, Michael P; De Koning, Harry P.

In: International Journal for Parasitology: Drugs and Drug Resistance, Vol. 7, No. 2, 01.08.2017, p. 206-226.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Functional and genetic evidence that nucleoside transport is highly conserved in Leishmania species

T2 - Implications for pyrimidine-based chemotherapy

AU - Alzahrani, Khalid J.H.

AU - Ali, Juma A M

AU - Eze, Anthonius A.

AU - Looi, Wan Limm

AU - Tagoe, Daniel N.A.

AU - Creek, Darren J.

AU - Barrett, Michael P

AU - De Koning, Harry P

PY - 2017/8/1

Y1 - 2017/8/1

N2 - 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.

AB - 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.

KW - 5-fluorouracil

KW - Leishmania

KW - Metabolomics

KW - Nucleoside transporter

KW - Pyrimidine chemotherapy

KW - Pyrimidine metabolism

KW - Uracil transporter

UR - http://www.scopus.com/inward/record.url?scp=85018661396&partnerID=8YFLogxK

U2 - 10.1016/j.ijpddr.2017.04.003

DO - 10.1016/j.ijpddr.2017.04.003

M3 - Article

VL - 7

SP - 206

EP - 226

JO - International Journal for Parasitology: Drugs and Drug Resistance

JF - International Journal for Parasitology: Drugs and Drug Resistance

SN - 2211-3207

IS - 2

ER -