A high susceptibility to redox imbalance of the transmissible stages of Plasmodium falciparum revealed with a luciferase-based mature gametocyte assay

Giulia Siciliano, T. R. Santha Kumar, Roberta Bona, Grazia Camarda, Maria Maddalena Calabretta, Luca Cevenini, Elisabeth Davioud-Charvet, Katja Becker, Andrea Cara, David A. Fidock, Pietro Alano

Research output: Contribution to journalArticle

Abstract

The goal to prevent Plasmodium falciparum transmission from humans to mosquitoes requires the identification of targetable metabolic processes in the mature (stage V) gametocytes, the sexual stages circulating in the bloodstream. This task is complicated by the apparently low metabolism of these cells, which renders them refractory to most antimalarial inhibitors and constrains the development of specific and sensitive cell-based assays. Here, we identify and functionally characterize the regulatory regions of the P. falciparum gene PF3D7_1234700, encoding a CPW-WPC protein and named here Upregulated in Late Gametocytes (ULG8), which we have leveraged to express reporter genes in mature male and female gametocytes. Using transgenic parasites containing a pfULG8-luciferase cassette, we investigated the susceptibility of stage V gametocytes to compounds specifically affecting redox metabolism. Our results reveal a high sensitivity of mature gametocytes to the glutathione reductase inhibitor and redox cycler drug methylene blue (MB). Using isobologram analysis, we find that a concomitant inhibition of the parasite enzyme glucose-6-phosphate dehydrogenase-6-phosphogluconolactonase, a key component of NADPH synthesis, potently synergizes MB activity. These data suggest that redox metabolism and detoxification activity play an unsuspected yet vital role in stage V gametocytes, rendering these cells exquisitely sensitive to decreases in NADPH concentration.

Original languageEnglish
Pages (from-to)306-318
Number of pages13
JournalMolecular Microbiology
Volume104
Issue number2
DOIs
Publication statusPublished - Apr 1 2017

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Plasmodium falciparum
Luciferases
Oxidation-Reduction
Methylene Blue
6-phosphogluconolactonase
NADP
Parasites
Glutathione Reductase
Glucosephosphate Dehydrogenase
Nucleic Acid Regulatory Sequences
Antimalarials
Culicidae
Reporter Genes
Enzymes
Pharmaceutical Preparations
Genes
Proteins

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

A high susceptibility to redox imbalance of the transmissible stages of Plasmodium falciparum revealed with a luciferase-based mature gametocyte assay. / Siciliano, Giulia; Santha Kumar, T. R.; Bona, Roberta; Camarda, Grazia; Calabretta, Maria Maddalena; Cevenini, Luca; Davioud-Charvet, Elisabeth; Becker, Katja; Cara, Andrea; Fidock, David A.; Alano, Pietro.

In: Molecular Microbiology, Vol. 104, No. 2, 01.04.2017, p. 306-318.

Research output: Contribution to journalArticle

Siciliano, G, Santha Kumar, TR, Bona, R, Camarda, G, Calabretta, MM, Cevenini, L, Davioud-Charvet, E, Becker, K, Cara, A, Fidock, DA & Alano, P 2017, 'A high susceptibility to redox imbalance of the transmissible stages of Plasmodium falciparum revealed with a luciferase-based mature gametocyte assay', Molecular Microbiology, vol. 104, no. 2, pp. 306-318. https://doi.org/10.1111/mmi.13626
Siciliano, Giulia ; Santha Kumar, T. R. ; Bona, Roberta ; Camarda, Grazia ; Calabretta, Maria Maddalena ; Cevenini, Luca ; Davioud-Charvet, Elisabeth ; Becker, Katja ; Cara, Andrea ; Fidock, David A. ; Alano, Pietro. / A high susceptibility to redox imbalance of the transmissible stages of Plasmodium falciparum revealed with a luciferase-based mature gametocyte assay. In: Molecular Microbiology. 2017 ; Vol. 104, No. 2. pp. 306-318.
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