Targeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules
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-...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2019
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/88939 http://hdl.handle.net/10220/48346 |
| _version_ | 1811694138956972032 |
|---|---|
| author | Preiser, Peter Rainer Reddy, D. Srinivasa Tan, Kevin S. W. Shanmugam, Dhanasekaran Chandramohanadas, Rajesh Subramanian, Gowtham Belekar, Meenakshi A. Shukla, Anurag Tong, Jie Xin Sinha, Ameya Chu, Trang T. T. Kulkarni, Akshay S. |
| author2 | Sullivan, William J. |
| author_facet | Sullivan, William J. Preiser, Peter Rainer Reddy, D. Srinivasa Tan, Kevin S. W. Shanmugam, Dhanasekaran Chandramohanadas, Rajesh Subramanian, Gowtham Belekar, Meenakshi A. Shukla, Anurag Tong, Jie Xin Sinha, Ameya Chu, Trang T. T. Kulkarni, Akshay S. |
| author_sort | Preiser, Peter Rainer |
| collection | NTU |
| description | 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. |
| first_indexed | 2024-10-01T07:02:49Z |
| format | Journal Article |
| id | ntu-10356/88939 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:02:49Z |
| publishDate | 2019 |
| record_format | dspace |
| spelling | ntu-10356/889392023-02-28T17:00:01Z Targeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules Preiser, Peter Rainer Reddy, D. Srinivasa Tan, Kevin S. W. Shanmugam, Dhanasekaran Chandramohanadas, Rajesh Subramanian, Gowtham Belekar, Meenakshi A. Shukla, Anurag Tong, Jie Xin Sinha, Ameya Chu, Trang T. T. Kulkarni, Akshay S. Sullivan, William J. School of Biological Sciences DRNTU::Science::Biological sciences Malaria MMV Malaria Box 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. ASTAR (Agency for Sci., Tech. and Research, S’pore) MOE (Min. of Education, S’pore) Published version 2019-05-23T07:50:51Z 2019-12-06T17:14:10Z 2019-05-23T07:50:51Z 2019-12-06T17:14:10Z 2018 Journal Article Subramanian, G., Belekar, M. A., Shukla, A., Tong, J. X., Sinha, A., Chu, T. T. T., . . . Chandramohanadas, R. (2018). Targeted Phenotypic Screening in Plasmodium falciparum and Toxoplasma gondii Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules. mSphere, 3(1), e00534-17-. doi:10.1128/mSphere.00534-17 https://hdl.handle.net/10356/88939 http://hdl.handle.net/10220/48346 10.1128/mSphere.00534-17 en mSphere © 2018 Subramanian et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. 22 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences Malaria MMV Malaria Box Preiser, Peter Rainer Reddy, D. Srinivasa Tan, Kevin S. W. Shanmugam, Dhanasekaran Chandramohanadas, Rajesh Subramanian, Gowtham Belekar, Meenakshi A. Shukla, Anurag Tong, Jie Xin Sinha, Ameya Chu, Trang T. T. Kulkarni, Akshay S. Targeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules |
| title | Targeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules |
| title_full | Targeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules |
| title_fullStr | Targeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules |
| title_full_unstemmed | Targeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules |
| title_short | Targeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules |
| title_sort | targeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules |
| topic | DRNTU::Science::Biological sciences Malaria MMV Malaria Box |
| url | https://hdl.handle.net/10356/88939 http://hdl.handle.net/10220/48346 |
| work_keys_str_mv | AT preiserpeterrainer targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT reddydsrinivasa targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT tankevinsw targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT shanmugamdhanasekaran targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT chandramohanadasrajesh targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT subramaniangowtham targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT belekarmeenakshia targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT shuklaanurag targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT tongjiexin targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT sinhaameya targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT chutrangtt targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules AT kulkarniakshays targetedphenotypicscreeninginplasmodiumfalciparumandtoxoplasmagondiirevealsnovelmodesofactionofmedicinesformalariaventuremalariaboxmolecules |