Targeted Phenotypic Screening in <named-content content-type="genus-species">Plasmodium falciparum</named-content> and <named-content content-type="genus-species">Toxoplasma gondii</named-content> Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules

ABSTRACT The Malaria Box collection includes 400 chemically diverse small molecules with documented potency against malaria parasite growth, but the underlying modes of action are largely unknown. Using complementary phenotypic screens against Plasmodium falciparum and Toxoplasma gondii, we report p...

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Autores principales:	Gowtham Subramanian, Meenakshi A. Belekar, Anurag Shukla, Jie Xin Tong, Ameya Sinha, Trang T. T. Chu, Akshay S. Kulkarni, Peter R. Preiser, D. Srinivasa Reddy, Kevin S. W. Tan, Dhanasekaran Shanmugam, Rajesh Chandramohanadas
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	MMV Malaria Box malaria Plasmodium falciparum Toxoplasma gondii apicoplast chemical phenotyping Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/d819bcc1aa584947876cb9484e9bb1d1
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spelling	oai:doaj.org-article:d819bcc1aa584947876cb9484e9bb1d12021-11-15T15:22:01ZTargeted Phenotypic Screening in <named-content content-type="genus-species">Plasmodium falciparum</named-content> and <named-content content-type="genus-species">Toxoplasma gondii</named-content> Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules10.1128/mSphere.00534-172379-5042https://doaj.org/article/d819bcc1aa584947876cb9484e9bb1d12018-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00534-17https://doaj.org/toc/2379-5042ABSTRACT The Malaria Box collection includes 400 chemically diverse small molecules with documented potency against malaria parasite growth, but the underlying modes of action are largely unknown. Using complementary phenotypic screens against Plasmodium falciparum and Toxoplasma gondii, we report phenotype-specific hits based on inhibition of overall parasite growth, apicoplast segregation, and egress or host invasion, providing hitherto unavailable insights into the possible mechanisms affected. First, the Malaria Box library was screened against tachyzoite stage T. gondii and the half-maximal effective concentrations (EC50s) of molecules showing ≥80% growth inhibition at 10 µM were determined. Comparison of the EC50s for T. gondii and P. falciparum identified a subset of 24 molecules with nanomolar potency against both parasites. Thirty molecules that failed to induce acute growth inhibition in T. gondii tachyzoites in a 2-day assay caused delayed parasite death upon extended exposure, with at least three molecules interfering with apicoplast segregation during daughter cell formation. Using flow cytometry and microscopy-based examinations, we prioritized 26 molecules with the potential to inhibit host cell egress/invasion during asexual developmental stages of P. falciparum. None of the inhibitors affected digestive vacuole integrity, ruling out a mechanism mediated by broadly specific protease inhibitor activity. Interestingly, five of the plasmodial egress inhibitors inhibited ionophore-induced egress of T. gondii tachyzoites. These findings highlight the advantage of comparative and targeted phenotypic screens in related species as a means to identify lead molecules with a conserved mode of action. Further work on target identification and mechanism analysis will facilitate the development of antiparasitic compounds with cross-species efficacy. IMPORTANCE The phylum Apicomplexa includes many human and animal pathogens, such as Plasmodium falciparum (human malaria) and Toxoplasma gondii (human and animal toxoplasmosis). Widespread resistance to current antimalarials and the lack of a commercial vaccine necessitate novel pharmacological interventions with distinct modes of action against malaria. For toxoplasmosis, new drugs to effectively eliminate tissue-dwelling latent cysts of the parasite are needed. The Malaria Box antimalarial collection, managed and distributed by the Medicines for Malaria Venture, includes molecules of novel chemical classes with proven antimalarial efficacy. Using targeted phenotypic assays of P. falciparum and T. gondii, we have identified a subset of the Malaria Box molecules as potent inhibitors of plastid segregation and parasite invasion and egress, thereby providing early insights into their probable mode of action. Five molecules that inhibit the egress of both parasites have been identified for further mechanistic studies. Thus, the approach we have used to identify novel molecules with defined modes of action in multiple parasites can expedite the development of pan-active antiparasitic agents.Gowtham SubramanianMeenakshi A. BelekarAnurag ShuklaJie Xin TongAmeya SinhaTrang T. T. ChuAkshay S. KulkarniPeter R. PreiserD. Srinivasa ReddyKevin S. W. TanDhanasekaran ShanmugamRajesh ChandramohanadasAmerican Society for MicrobiologyarticleMMV Malaria BoxmalariaPlasmodium falciparumToxoplasma gondiiapicoplastchemical phenotypingMicrobiologyQR1-502ENmSphere, Vol 3, Iss 1 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	MMV Malaria Box malaria Plasmodium falciparum Toxoplasma gondii apicoplast chemical phenotyping Microbiology QR1-502
spellingShingle	MMV Malaria Box malaria Plasmodium falciparum Toxoplasma gondii apicoplast chemical phenotyping Microbiology QR1-502 Gowtham Subramanian Meenakshi A. Belekar Anurag Shukla Jie Xin Tong Ameya Sinha Trang T. T. Chu Akshay S. Kulkarni Peter R. Preiser D. Srinivasa Reddy Kevin S. W. Tan Dhanasekaran Shanmugam Rajesh Chandramohanadas Targeted Phenotypic Screening in <named-content content-type="genus-species">Plasmodium falciparum</named-content> and <named-content content-type="genus-species">Toxoplasma gondii</named-content> Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules
description	ABSTRACT The Malaria Box collection includes 400 chemically diverse small molecules with documented potency against malaria parasite growth, but the underlying modes of action are largely unknown. Using complementary phenotypic screens against Plasmodium falciparum and Toxoplasma gondii, we report phenotype-specific hits based on inhibition of overall parasite growth, apicoplast segregation, and egress or host invasion, providing hitherto unavailable insights into the possible mechanisms affected. First, the Malaria Box library was screened against tachyzoite stage T. gondii and the half-maximal effective concentrations (EC50s) of molecules showing ≥80% growth inhibition at 10 µM were determined. Comparison of the EC50s for T. gondii and P. falciparum identified a subset of 24 molecules with nanomolar potency against both parasites. Thirty molecules that failed to induce acute growth inhibition in T. gondii tachyzoites in a 2-day assay caused delayed parasite death upon extended exposure, with at least three molecules interfering with apicoplast segregation during daughter cell formation. Using flow cytometry and microscopy-based examinations, we prioritized 26 molecules with the potential to inhibit host cell egress/invasion during asexual developmental stages of P. falciparum. None of the inhibitors affected digestive vacuole integrity, ruling out a mechanism mediated by broadly specific protease inhibitor activity. Interestingly, five of the plasmodial egress inhibitors inhibited ionophore-induced egress of T. gondii tachyzoites. These findings highlight the advantage of comparative and targeted phenotypic screens in related species as a means to identify lead molecules with a conserved mode of action. Further work on target identification and mechanism analysis will facilitate the development of antiparasitic compounds with cross-species efficacy. IMPORTANCE The phylum Apicomplexa includes many human and animal pathogens, such as Plasmodium falciparum (human malaria) and Toxoplasma gondii (human and animal toxoplasmosis). Widespread resistance to current antimalarials and the lack of a commercial vaccine necessitate novel pharmacological interventions with distinct modes of action against malaria. For toxoplasmosis, new drugs to effectively eliminate tissue-dwelling latent cysts of the parasite are needed. The Malaria Box antimalarial collection, managed and distributed by the Medicines for Malaria Venture, includes molecules of novel chemical classes with proven antimalarial efficacy. Using targeted phenotypic assays of P. falciparum and T. gondii, we have identified a subset of the Malaria Box molecules as potent inhibitors of plastid segregation and parasite invasion and egress, thereby providing early insights into their probable mode of action. Five molecules that inhibit the egress of both parasites have been identified for further mechanistic studies. Thus, the approach we have used to identify novel molecules with defined modes of action in multiple parasites can expedite the development of pan-active antiparasitic agents.
format	article
author	Gowtham Subramanian Meenakshi A. Belekar Anurag Shukla Jie Xin Tong Ameya Sinha Trang T. T. Chu Akshay S. Kulkarni Peter R. Preiser D. Srinivasa Reddy Kevin S. W. Tan Dhanasekaran Shanmugam Rajesh Chandramohanadas
author_facet	Gowtham Subramanian Meenakshi A. Belekar Anurag Shukla Jie Xin Tong Ameya Sinha Trang T. T. Chu Akshay S. Kulkarni Peter R. Preiser D. Srinivasa Reddy Kevin S. W. Tan Dhanasekaran Shanmugam Rajesh Chandramohanadas
author_sort	Gowtham Subramanian
title	Targeted Phenotypic Screening in <named-content content-type="genus-species">Plasmodium falciparum</named-content> and <named-content content-type="genus-species">Toxoplasma gondii</named-content> Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules
title_short	Targeted Phenotypic Screening in <named-content content-type="genus-species">Plasmodium falciparum</named-content> and <named-content content-type="genus-species">Toxoplasma gondii</named-content> Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules
title_full	Targeted Phenotypic Screening in <named-content content-type="genus-species">Plasmodium falciparum</named-content> and <named-content content-type="genus-species">Toxoplasma gondii</named-content> Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules
title_fullStr	Targeted Phenotypic Screening in <named-content content-type="genus-species">Plasmodium falciparum</named-content> and <named-content content-type="genus-species">Toxoplasma gondii</named-content> Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules
title_full_unstemmed	Targeted Phenotypic Screening in <named-content content-type="genus-species">Plasmodium falciparum</named-content> and <named-content content-type="genus-species">Toxoplasma gondii</named-content> Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules
title_sort	targeted phenotypic screening in <named-content content-type="genus-species">plasmodium falciparum</named-content> and <named-content content-type="genus-species">toxoplasma gondii</named-content> reveals novel modes of action of medicines for malaria venture malaria box molecules
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/d819bcc1aa584947876cb9484e9bb1d1
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Targeted Phenotypic Screening in <named-content content-type="genus-species">Plasmodium falciparum</named-content> and <named-content content-type="genus-species">Toxoplasma gondii</named-content> Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules

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