Genetic modulators of Plasmodium falciparum parasites asexual fitness and resistance to clinical and preclinical compounds
Emergence of malarial Plasmodium falciparum KEL1/PLA1 co-lineage in Southeast Asia caused the failure of dihydroartemisin+piperaquine (DHA+PPQ). PfKelch (k13) C580Y mutation and plasmepsins 2/3 gene (pm2/3) amplification, markers of DHA and PPQ resistance respectively, are hallmarks of KEL1/PLA1 par...
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| Format: | Final Year Project (FYP) |
| Language: | English |
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Nanyang Technological University
2023
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| Online Access: | https://hdl.handle.net/10356/167504 |
| _version_ | 1826117912818089984 |
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| author | Hong, Davin |
| author2 | - |
| author_facet | - Hong, Davin |
| author_sort | Hong, Davin |
| collection | NTU |
| description | Emergence of malarial Plasmodium falciparum KEL1/PLA1 co-lineage in Southeast Asia caused the failure of dihydroartemisin+piperaquine (DHA+PPQ). PfKelch (k13) C580Y mutation and plasmepsins 2/3 gene (pm2/3) amplification, markers of DHA and PPQ resistance respectively, are hallmarks of KEL1/PLA1 parasites such as RF7, while PPQ resistance marker, M343L mutation in pfcrt on a mutant pfcrt Dd2 background, also emerged. Interactions between pfcrt M343L mutation and multicopy pm2/3 to confer PPQ resistance and the impact of KEL1/PLA1 hallmarks on parasite asexual fitness has not been elucidated. By using CRISPR/Cas9 editing and limiting dilution cloning of RF7 parasite and its drug-sensitive NF54 cross progeny, we illustrate the importance of pfcrt M343L background in PPQ resistance with pm2/3 amplification augmenting the phenotype. M343L mutation, however, did not confer resistance to artemisinin partner drugs lumefantrine or mefloquine, while pm2/3 amplification augmented resistance phenotype to preclinical compound MMV675939. Drug-free long-term competitive fitness assays between RF7 clones revealed that multicopy pm2/3 on mutant k13 led to moderate parasite fitness defect. These results confirmed DHA+PPQ selects for pm2/3 amplification and mutant k13/pfcrt parasites, allowing RF7 to emerge, but poor fitness in absence of PPQ can deamplify pm2/3. Current parasite genotypes reinforced the appeal of artesunate+mefloquine as a DHA+PPQ replacement. |
| first_indexed | 2024-10-01T04:35:08Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/167504 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T04:35:08Z |
| publishDate | 2023 |
| publisher | Nanyang Technological University |
| record_format | dspace |
| spelling | ntu-10356/1675042023-05-29T15:33:07Z Genetic modulators of Plasmodium falciparum parasites asexual fitness and resistance to clinical and preclinical compounds Hong, Davin - School of Biological Sciences Columbia University David A. Fidock df2260@cumc.columbia.edu Science::Biological sciences::Microbiology::Drug Resistance Science::Biological sciences::Genetics Emergence of malarial Plasmodium falciparum KEL1/PLA1 co-lineage in Southeast Asia caused the failure of dihydroartemisin+piperaquine (DHA+PPQ). PfKelch (k13) C580Y mutation and plasmepsins 2/3 gene (pm2/3) amplification, markers of DHA and PPQ resistance respectively, are hallmarks of KEL1/PLA1 parasites such as RF7, while PPQ resistance marker, M343L mutation in pfcrt on a mutant pfcrt Dd2 background, also emerged. Interactions between pfcrt M343L mutation and multicopy pm2/3 to confer PPQ resistance and the impact of KEL1/PLA1 hallmarks on parasite asexual fitness has not been elucidated. By using CRISPR/Cas9 editing and limiting dilution cloning of RF7 parasite and its drug-sensitive NF54 cross progeny, we illustrate the importance of pfcrt M343L background in PPQ resistance with pm2/3 amplification augmenting the phenotype. M343L mutation, however, did not confer resistance to artemisinin partner drugs lumefantrine or mefloquine, while pm2/3 amplification augmented resistance phenotype to preclinical compound MMV675939. Drug-free long-term competitive fitness assays between RF7 clones revealed that multicopy pm2/3 on mutant k13 led to moderate parasite fitness defect. These results confirmed DHA+PPQ selects for pm2/3 amplification and mutant k13/pfcrt parasites, allowing RF7 to emerge, but poor fitness in absence of PPQ can deamplify pm2/3. Current parasite genotypes reinforced the appeal of artesunate+mefloquine as a DHA+PPQ replacement. Bachelor of Science in Biological Sciences 2023-05-29T08:49:00Z 2023-05-29T08:49:00Z 2023 Final Year Project (FYP) Hong, D. (2023). Genetic modulators of Plasmodium falciparum parasites asexual fitness and resistance to clinical and preclinical compounds. Final Year Project (FYP), Nanyang Technological University, Singapore. https://hdl.handle.net/10356/167504 https://hdl.handle.net/10356/167504 en application/pdf Nanyang Technological University |
| spellingShingle | Science::Biological sciences::Microbiology::Drug Resistance Science::Biological sciences::Genetics Hong, Davin Genetic modulators of Plasmodium falciparum parasites asexual fitness and resistance to clinical and preclinical compounds |
| title | Genetic modulators of Plasmodium falciparum parasites asexual fitness and resistance to clinical and preclinical compounds |
| title_full | Genetic modulators of Plasmodium falciparum parasites asexual fitness and resistance to clinical and preclinical compounds |
| title_fullStr | Genetic modulators of Plasmodium falciparum parasites asexual fitness and resistance to clinical and preclinical compounds |
| title_full_unstemmed | Genetic modulators of Plasmodium falciparum parasites asexual fitness and resistance to clinical and preclinical compounds |
| title_short | Genetic modulators of Plasmodium falciparum parasites asexual fitness and resistance to clinical and preclinical compounds |
| title_sort | genetic modulators of plasmodium falciparum parasites asexual fitness and resistance to clinical and preclinical compounds |
| topic | Science::Biological sciences::Microbiology::Drug Resistance Science::Biological sciences::Genetics |
| url | https://hdl.handle.net/10356/167504 |
| work_keys_str_mv | AT hongdavin geneticmodulatorsofplasmodiumfalciparumparasitesasexualfitnessandresistancetoclinicalandpreclinicalcompounds |