Nowhere to hide : interrogating different metabolic parameters of Plasmodium falciparum gametocytes in a transmission blocking drug discovery pipeline towards malaria elimination

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dc.contributor.author Reader, Janette
dc.contributor.author Botha, M.E. (Mariette)
dc.contributor.author Theron, Anjo
dc.contributor.author Lauterbach, Sonja B.
dc.contributor.author Rossouw, C.L. (Claire Louise)
dc.contributor.author Engelbrecht, Dewaldt
dc.contributor.author Wepener, Melanie
dc.contributor.author Smit, Annél
dc.contributor.author Leroy, Didier
dc.contributor.author Mancama, Dalu
dc.contributor.author Coetzer, Theresa L.
dc.contributor.author Birkholtz, Lyn-Marie
dc.date.accessioned 2015-06-25T10:22:11Z
dc.date.available 2015-06-25T10:22:11Z
dc.date.issued 2015
dc.description.abstract BACKGROUND : The discovery of malaria transmission-blocking compounds is seen as key to malaria elimination strategies and gametocyte-screening platforms are critical filters to identify active molecules. However, unlike asexual parasite assays measuring parasite proliferation, greater variability in end-point readout exists between different gametocytocidal assays. This is compounded by difficulties in routinely producing viable, functional and stage-specific gametocyte populations. Here, a parallel evaluation of four assay platforms on the same gametocyte populations was performed for the first time. This allowed the direct comparison of the ability of different assay platforms to detect compounds with gametocytocidal activity and revealed caveats in some assay readouts that interrogate different parasite biological functions. METHODS : Gametocytogenesis from Plasmodium falciparum (NF54) was optimized with a robust and standardized protocol. ATP, pLDH, luciferase reporter and PrestoBlue® assays were compared in context of a set of 10 reference compounds. The assays were performed in parallel on the same gametocyte preparation (except for luciferase reporter lines) using the same drug preparations (48 h). The remaining parameters for each assay were all comparable. RESULTS : A highly robust method for generating viable and functional gametocytes was developed and comprehensively validated resulting in an average gametocytaemia of 4 %. Subsequent parallel assays for gametocytocidal activity indicated that different assay platforms were not able to screen compounds with variant chemical scaffolds similarly. Luciferase reporter assays revealed that synchronized stage-specific gametocyte production is essential for drug discovery, as differential susceptibility in various gametocyte developmental populations is evident. CONCLUSIONS : With this study, the key parameters for assays aiming at testing the gametocytocidal activity of potential transmission blocking molecules against Plasmodium gametocytes were accurately dissected. This first and uniquely comparative study emphasizes differential effects seen with the use of different assay platforms interrogating variant biological systems. Whilst this data is informative from a biological perspective and may provide indications of the drug mode of action, it does highlight the care that must be taken when screening broaddiversity chemotypes with a single assay platform against gametocytes for which the biology is not clearly understood. en_ZA
dc.description.librarian hb2015 en_ZA
dc.description.sponsorship South African Medical Research Council Strategic Health Initiatives Partnerships with the Medicines for Malaria Venture as well as the Council for Scientific and Industrial Research, and the 3R Foundation (project 118–10). en_ZA
dc.description.uri http://www.malariajournal.com en_ZA
dc.identifier.citation Reader, J, Botha, M, Theron, A, Lauterbach, SB, Rossouw, C, Engelbrecht, D, Wepener, M, Smit, A, Leroy, D, Mancama, D, Coetzer, TL & Birkholtz, LM 2015, 'Nowhere to hide : interrogating different metabolic parameters of Plasmodium falciparum gametocytes in a transmission blocking drug discovery pipeline towards malaria elimination', Malaria Journal, vol. 14, art. no. 213, pp. 1-17. en_ZA
dc.identifier.issn 1475-2875
dc.identifier.other 10.1186/s12936-015-0718-z
dc.identifier.uri http://hdl.handle.net/2263/45785
dc.language.iso en en_ZA
dc.publisher BioMed Central en_ZA
dc.rights © 2015 Reader et al.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0). en_ZA
dc.subject Malaria en_ZA
dc.subject Plasmodium falciparum en_ZA
dc.subject Gametocyte en_ZA
dc.subject Gametocytocidal compounds en_ZA
dc.subject Transmissionblocking assays en_ZA
dc.title Nowhere to hide : interrogating different metabolic parameters of Plasmodium falciparum gametocytes in a transmission blocking drug discovery pipeline towards malaria elimination en_ZA
dc.type Article en_ZA


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