A high-content phenotypic screen reveals the disruptive potency of quinacrine and 3',4'-Dichlorobenzamil on the digestive vacuole of plasmodium falciparum

Plasmodium falciparum is the etiological agent of malignant malaria and has been shown to exhibit features resembling programmed cell death. This is triggered upon treatment with low micromolar doses of chloroquine or other lysosomotrophic compounds and is associated with leakage of the digestive va...

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Main Authors: Lee, Y., Goh, A., Ch'Ng, J., Nosten, F., Preiser, P., Pervaiz, Shazib, Yadav, S., Tan, K.
Format: Journal Article
Published: American Society for Microbiology 2014
Online Access:http://hdl.handle.net/20.500.11937/51046
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author Lee, Y.
Goh, A.
Ch'Ng, J.
Nosten, F.
Preiser, P.
Pervaiz, Shazib
Yadav, S.
Tan, K.
author_facet Lee, Y.
Goh, A.
Ch'Ng, J.
Nosten, F.
Preiser, P.
Pervaiz, Shazib
Yadav, S.
Tan, K.
author_sort Lee, Y.
building Curtin Institutional Repository
collection Online Access
description Plasmodium falciparum is the etiological agent of malignant malaria and has been shown to exhibit features resembling programmed cell death. This is triggered upon treatment with low micromolar doses of chloroquine or other lysosomotrophic compounds and is associated with leakage of the digestive vacuole contents. In order to exploit this cell death pathway, we developed a high-content screening method to select compounds that can disrupt the parasite vacuole, as measured by the leakage of intravacuolar Ca2+. This assay uses the ImageStream 100, an imaging-capable flow cytometer, to assess the distribution of the fluorescent calcium probe Fluo-4. We obtained two hits from a small library of 25 test compounds, quinacrine and 3',4'-dichlorobenzamil. The ability of these compounds to permeabilize the digestive vacuole in laboratory strains and clinical isolates was validated by confocal microscopy. The hits could induce programmed cell death features in both chloroquine-sensitive and -resistant laboratory strains. Quinacrine was effective at inhibiting field isolates in a 48-h reinvasion assay regardless of artemisinin clearance status. We therefore present as proof of concept a phenotypic screening method with the potential to provide mechanistic insights to the activity of antimalarial drugs. © 2014, American Society for Microbiology.
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spelling curtin-20.500.11937-510462017-09-13T15:34:26Z A high-content phenotypic screen reveals the disruptive potency of quinacrine and 3',4'-Dichlorobenzamil on the digestive vacuole of plasmodium falciparum Lee, Y. Goh, A. Ch'Ng, J. Nosten, F. Preiser, P. Pervaiz, Shazib Yadav, S. Tan, K. Plasmodium falciparum is the etiological agent of malignant malaria and has been shown to exhibit features resembling programmed cell death. This is triggered upon treatment with low micromolar doses of chloroquine or other lysosomotrophic compounds and is associated with leakage of the digestive vacuole contents. In order to exploit this cell death pathway, we developed a high-content screening method to select compounds that can disrupt the parasite vacuole, as measured by the leakage of intravacuolar Ca2+. This assay uses the ImageStream 100, an imaging-capable flow cytometer, to assess the distribution of the fluorescent calcium probe Fluo-4. We obtained two hits from a small library of 25 test compounds, quinacrine and 3',4'-dichlorobenzamil. The ability of these compounds to permeabilize the digestive vacuole in laboratory strains and clinical isolates was validated by confocal microscopy. The hits could induce programmed cell death features in both chloroquine-sensitive and -resistant laboratory strains. Quinacrine was effective at inhibiting field isolates in a 48-h reinvasion assay regardless of artemisinin clearance status. We therefore present as proof of concept a phenotypic screening method with the potential to provide mechanistic insights to the activity of antimalarial drugs. © 2014, American Society for Microbiology. 2014 Journal Article http://hdl.handle.net/20.500.11937/51046 10.1128/AAC.01441-13 American Society for Microbiology unknown
spellingShingle Lee, Y.
Goh, A.
Ch'Ng, J.
Nosten, F.
Preiser, P.
Pervaiz, Shazib
Yadav, S.
Tan, K.
A high-content phenotypic screen reveals the disruptive potency of quinacrine and 3',4'-Dichlorobenzamil on the digestive vacuole of plasmodium falciparum
title A high-content phenotypic screen reveals the disruptive potency of quinacrine and 3',4'-Dichlorobenzamil on the digestive vacuole of plasmodium falciparum
title_full A high-content phenotypic screen reveals the disruptive potency of quinacrine and 3',4'-Dichlorobenzamil on the digestive vacuole of plasmodium falciparum
title_fullStr A high-content phenotypic screen reveals the disruptive potency of quinacrine and 3',4'-Dichlorobenzamil on the digestive vacuole of plasmodium falciparum
title_full_unstemmed A high-content phenotypic screen reveals the disruptive potency of quinacrine and 3',4'-Dichlorobenzamil on the digestive vacuole of plasmodium falciparum
title_short A high-content phenotypic screen reveals the disruptive potency of quinacrine and 3',4'-Dichlorobenzamil on the digestive vacuole of plasmodium falciparum
title_sort high-content phenotypic screen reveals the disruptive potency of quinacrine and 3',4'-dichlorobenzamil on the digestive vacuole of plasmodium falciparum
url http://hdl.handle.net/20.500.11937/51046