Evidence for Regulation of Hemoglobin Metabolism and Intracellular Ionic Flux by the Plasmodium falciparum Chloroquine Resistance Transporter

Andrew H. Lee, Satish K. Dhingra, Ian A. Lewis, Maneesh K. Singh, Amila Siriwardana, Seema Dalal, Kelly Rubiano, Matthias S. Klein, Katelynn S. Baska, Sanjeev Krishna, Michael Klemba, Paul D. Roepe, Manuel Llinás, Celia R.S. Garcia, David A. Fidock

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Plasmodium falciparum multidrug resistance constitutes a major obstacle to the global malaria elimination campaign. Specific mutations in the Plasmodium falciparum chloroquine resistance transporter (PfCRT) mediate resistance to the 4-aminoquinoline drug chloroquine and impact parasite susceptibility to several partner agents used in current artemisinin-based combination therapies, including amodiaquine. By examining gene-edited parasites, we report that the ability of the wide-spread Dd2 PfCRT isoform to mediate chloroquine and amodiaquine resistance is substantially reduced by the addition of the PfCRT L272F mutation, which arose under blasticidin selection. We also provide evidence that L272F confers a significant fitness cost to asexual blood stage parasites. Studies with amino acid-restricted media identify this mutant as a methionine auxotroph. Metabolomic analysis also reveals an accumulation of short, hemoglobin-derived peptides in the Dd2 + L272F and Dd2 isoforms, compared with parasites expressing wild-type PfCRT. Physiologic studies with the ionophores monensin and nigericin support an impact of PfCRT isoforms on Ca2+ release, with substantially reduced Ca2+ levels observed in Dd2 + L272F parasites. Our data reveal a central role for PfCRT in regulating hemoglobin catabolism, amino acid availability, and ionic balance in P. falciparum, in addition to its role in determining parasite susceptibility to heme-binding 4-aminoquinoline drugs.

Original languageEnglish (US)
Article number13578
JournalScientific reports
Volume8
Issue number1
DOIs
StatePublished - Dec 1 2018

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Parasites
Hemoglobins
Amodiaquine
Protein Isoforms
Chloroquine
Plasmodium falciparum
Nigericin
Amino Acids
Monensin
Mutation
Metabolomics
Ionophores
Multiple Drug Resistance
Heme
Pharmaceutical Preparations
Methionine
Malaria
Plasmodium falciparum PfCRT protein
Costs and Cost Analysis
Peptides

All Science Journal Classification (ASJC) codes

  • General

Cite this

Lee, Andrew H. ; Dhingra, Satish K. ; Lewis, Ian A. ; Singh, Maneesh K. ; Siriwardana, Amila ; Dalal, Seema ; Rubiano, Kelly ; Klein, Matthias S. ; Baska, Katelynn S. ; Krishna, Sanjeev ; Klemba, Michael ; Roepe, Paul D. ; Llinás, Manuel ; Garcia, Celia R.S. ; Fidock, David A. / Evidence for Regulation of Hemoglobin Metabolism and Intracellular Ionic Flux by the Plasmodium falciparum Chloroquine Resistance Transporter. In: Scientific reports. 2018 ; Vol. 8, No. 1.
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abstract = "Plasmodium falciparum multidrug resistance constitutes a major obstacle to the global malaria elimination campaign. Specific mutations in the Plasmodium falciparum chloroquine resistance transporter (PfCRT) mediate resistance to the 4-aminoquinoline drug chloroquine and impact parasite susceptibility to several partner agents used in current artemisinin-based combination therapies, including amodiaquine. By examining gene-edited parasites, we report that the ability of the wide-spread Dd2 PfCRT isoform to mediate chloroquine and amodiaquine resistance is substantially reduced by the addition of the PfCRT L272F mutation, which arose under blasticidin selection. We also provide evidence that L272F confers a significant fitness cost to asexual blood stage parasites. Studies with amino acid-restricted media identify this mutant as a methionine auxotroph. Metabolomic analysis also reveals an accumulation of short, hemoglobin-derived peptides in the Dd2 + L272F and Dd2 isoforms, compared with parasites expressing wild-type PfCRT. Physiologic studies with the ionophores monensin and nigericin support an impact of PfCRT isoforms on Ca2+ release, with substantially reduced Ca2+ levels observed in Dd2 + L272F parasites. Our data reveal a central role for PfCRT in regulating hemoglobin catabolism, amino acid availability, and ionic balance in P. falciparum, in addition to its role in determining parasite susceptibility to heme-binding 4-aminoquinoline drugs.",
author = "Lee, {Andrew H.} and Dhingra, {Satish K.} and Lewis, {Ian A.} and Singh, {Maneesh K.} and Amila Siriwardana and Seema Dalal and Kelly Rubiano and Klein, {Matthias S.} and Baska, {Katelynn S.} and Sanjeev Krishna and Michael Klemba and Roepe, {Paul D.} and Manuel Llin{\'a}s and Garcia, {Celia R.S.} and Fidock, {David A.}",
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Lee, AH, Dhingra, SK, Lewis, IA, Singh, MK, Siriwardana, A, Dalal, S, Rubiano, K, Klein, MS, Baska, KS, Krishna, S, Klemba, M, Roepe, PD, Llinás, M, Garcia, CRS & Fidock, DA 2018, 'Evidence for Regulation of Hemoglobin Metabolism and Intracellular Ionic Flux by the Plasmodium falciparum Chloroquine Resistance Transporter', Scientific reports, vol. 8, no. 1, 13578. https://doi.org/10.1038/s41598-018-31715-9

Evidence for Regulation of Hemoglobin Metabolism and Intracellular Ionic Flux by the Plasmodium falciparum Chloroquine Resistance Transporter. / Lee, Andrew H.; Dhingra, Satish K.; Lewis, Ian A.; Singh, Maneesh K.; Siriwardana, Amila; Dalal, Seema; Rubiano, Kelly; Klein, Matthias S.; Baska, Katelynn S.; Krishna, Sanjeev; Klemba, Michael; Roepe, Paul D.; Llinás, Manuel; Garcia, Celia R.S.; Fidock, David A.

In: Scientific reports, Vol. 8, No. 1, 13578, 01.12.2018.

Research output: Contribution to journalArticle

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T1 - Evidence for Regulation of Hemoglobin Metabolism and Intracellular Ionic Flux by the Plasmodium falciparum Chloroquine Resistance Transporter

AU - Lee, Andrew H.

AU - Dhingra, Satish K.

AU - Lewis, Ian A.

AU - Singh, Maneesh K.

AU - Siriwardana, Amila

AU - Dalal, Seema

AU - Rubiano, Kelly

AU - Klein, Matthias S.

AU - Baska, Katelynn S.

AU - Krishna, Sanjeev

AU - Klemba, Michael

AU - Roepe, Paul D.

AU - Llinás, Manuel

AU - Garcia, Celia R.S.

AU - Fidock, David A.

PY - 2018/12/1

Y1 - 2018/12/1

N2 - Plasmodium falciparum multidrug resistance constitutes a major obstacle to the global malaria elimination campaign. Specific mutations in the Plasmodium falciparum chloroquine resistance transporter (PfCRT) mediate resistance to the 4-aminoquinoline drug chloroquine and impact parasite susceptibility to several partner agents used in current artemisinin-based combination therapies, including amodiaquine. By examining gene-edited parasites, we report that the ability of the wide-spread Dd2 PfCRT isoform to mediate chloroquine and amodiaquine resistance is substantially reduced by the addition of the PfCRT L272F mutation, which arose under blasticidin selection. We also provide evidence that L272F confers a significant fitness cost to asexual blood stage parasites. Studies with amino acid-restricted media identify this mutant as a methionine auxotroph. Metabolomic analysis also reveals an accumulation of short, hemoglobin-derived peptides in the Dd2 + L272F and Dd2 isoforms, compared with parasites expressing wild-type PfCRT. Physiologic studies with the ionophores monensin and nigericin support an impact of PfCRT isoforms on Ca2+ release, with substantially reduced Ca2+ levels observed in Dd2 + L272F parasites. Our data reveal a central role for PfCRT in regulating hemoglobin catabolism, amino acid availability, and ionic balance in P. falciparum, in addition to its role in determining parasite susceptibility to heme-binding 4-aminoquinoline drugs.

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