Amodiaquine drug pressure selects nonsynonymous mutations in pantothenate kinase 1, diacylglycerol kinase, and phosphatidylinositol-4 kinase in Plasmodium berghei ANKA [version 3; peer review: 1 approved, 3 approved with reservations, 1 not approved]
Background Lumefantrine (LM), piperaquine (PQ), and amodiaquine (AQ), the long-acting components of the artemisinin-based combination therapies (ACTs), are a cornerstone of malaria treatment in Africa. Studies have shown that PQ, AQ, and LM resistance may arise independently of predicted modes of ac...
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F1000 Research Ltd
2023-10-01
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| Online Access: | https://openresearchafrica.org/articles/5-28/v3 |
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| author | Peter Mwitari Francis Kimani Daniel Kiboi Brenda Muriithi Jean Chepngetich Kevin Thiong'o Beatrice Gachie Jeremiah Gathirwa Mercy Jepkorir |
| author_facet | Peter Mwitari Francis Kimani Daniel Kiboi Brenda Muriithi Jean Chepngetich Kevin Thiong'o Beatrice Gachie Jeremiah Gathirwa Mercy Jepkorir |
| author_sort | Peter Mwitari |
| collection | DOAJ |
| description | Background Lumefantrine (LM), piperaquine (PQ), and amodiaquine (AQ), the long-acting components of the artemisinin-based combination therapies (ACTs), are a cornerstone of malaria treatment in Africa. Studies have shown that PQ, AQ, and LM resistance may arise independently of predicted modes of action. Protein kinases have emerged as mediators of drug action and efficacy in malaria parasites; however, the link between top druggable Plasmodium kinases with LM, PQ, and AQ resistance remains unclear. Using LM, PQ, or AQ-resistant Plasmodium berghei parasites, we have evaluated the association of choline kinase (CK), pantothenate kinase 1 (PANK1), diacylglycerol kinase (DAGK), and phosphatidylinositol-4 kinase (PI4Kβ), and calcium-dependent protein kinase 1 (CDPK1) with LM, PQ, and AQ resistance in Plasmodium berghei ANKA. Methods We used in silico bioinformatics tools to identify ligand-binding motifs, active sites, and sequence conservation across the different parasites. We then used PCR and sequencing analysis to probe for single nucleotide polymorphisms (SNPs) within the predicted functional motifs in the CK, PANK1, DAGK, PI4Kβ, and CDPK1. Using qPCR analysis, we measured the mRNA amount of PANK1, DAGK, and PI4Kβ at trophozoites and schizonts stages. Results We reveal sequence conservation and unique ligand-binding motifs in the CK, PANK1, DAGK, PI4Kβ, and CDPK1 across malaria species. DAGK, PANK1, and PI4Kβ possessed nonsynonymous mutations; surprisingly, the mutations only occurred in the AQr parasites. PANK1 acquired Asn394His, while DAGK contained K270R and K292R mutations. PI4Kβ had Asp366Asn, Ser1367Arg, Tyr1394Asn and Asp1423Asn. We show downregulation of PANK1, DAGK, and PI4Kβ in the trophozoites but upregulation at the schizonts stages in the AQr parasites. Conclusions The selective acquisition of the mutations and the differential gene expression in AQ-resistant parasites may signify proteins under AQ pressure. The role of the mutations in the resistant parasites and their impact on drug responses require investigations using reverse genetics techniques in malaria parasites. |
| format | Article |
| id | doaj-art-c060ad9bd5eb4c4285dca6a31a995c0c |
| institution | OA Journals |
| issn | 2752-6925 |
| language | English |
| publishDate | 2023-10-01 |
| publisher | F1000 Research Ltd |
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| series | Open Research Africa |
| spelling | doaj-art-c060ad9bd5eb4c4285dca6a31a995c0c2025-08-20T01:58:54ZengF1000 Research LtdOpen Research Africa2752-69252023-10-01510.12688/openresafrica.13436.315632Amodiaquine drug pressure selects nonsynonymous mutations in pantothenate kinase 1, diacylglycerol kinase, and phosphatidylinositol-4 kinase in Plasmodium berghei ANKA [version 3; peer review: 1 approved, 3 approved with reservations, 1 not approved]Peter Mwitari0https://orcid.org/0000-0002-6634-0281Francis Kimani1https://orcid.org/0000-0001-5150-2068Daniel Kiboi2https://orcid.org/0000-0003-0797-7378Brenda Muriithi3Jean Chepngetich4Kevin Thiong'o5Beatrice Gachie6Jeremiah Gathirwa7Mercy Jepkorir8Centre for Traditional Medicine and Drug Research, Kenya Medical Research Institute, Nairobi, 54840, 00200, KenyaCentre for Biotechnology Research and Development, Kenya Medical Research Institute, Nairobi, 54840, 00200, KenyaDepartment of Biochemistry, Jomo Kenyatta University of Agriculture and Technology, Nairobi, 62000, 00200, KenyaCentre for Traditional Medicine and Drug Research, Kenya Medical Research Institute, Nairobi, 54840, 00200, KenyaDepartment of Molecular Biology and Biotechnology, Pan African University Institute for Basic Sciences, Technology and Innovation, Nairobi, 62000, 00200, KenyaCentre for Biotechnology Research and Development, Kenya Medical Research Institute, Nairobi, 54840, 00200, KenyaDepartment of Molecular Biology and Biotechnology, Pan African University Institute for Basic Sciences, Technology and Innovation, Nairobi, 62000, 00200, KenyaCentre for Traditional Medicine and Drug Research, Kenya Medical Research Institute, Nairobi, 54840, 00200, KenyaCentre for Traditional Medicine and Drug Research, Kenya Medical Research Institute, Nairobi, 54840, 00200, KenyaBackground Lumefantrine (LM), piperaquine (PQ), and amodiaquine (AQ), the long-acting components of the artemisinin-based combination therapies (ACTs), are a cornerstone of malaria treatment in Africa. Studies have shown that PQ, AQ, and LM resistance may arise independently of predicted modes of action. Protein kinases have emerged as mediators of drug action and efficacy in malaria parasites; however, the link between top druggable Plasmodium kinases with LM, PQ, and AQ resistance remains unclear. Using LM, PQ, or AQ-resistant Plasmodium berghei parasites, we have evaluated the association of choline kinase (CK), pantothenate kinase 1 (PANK1), diacylglycerol kinase (DAGK), and phosphatidylinositol-4 kinase (PI4Kβ), and calcium-dependent protein kinase 1 (CDPK1) with LM, PQ, and AQ resistance in Plasmodium berghei ANKA. Methods We used in silico bioinformatics tools to identify ligand-binding motifs, active sites, and sequence conservation across the different parasites. We then used PCR and sequencing analysis to probe for single nucleotide polymorphisms (SNPs) within the predicted functional motifs in the CK, PANK1, DAGK, PI4Kβ, and CDPK1. Using qPCR analysis, we measured the mRNA amount of PANK1, DAGK, and PI4Kβ at trophozoites and schizonts stages. Results We reveal sequence conservation and unique ligand-binding motifs in the CK, PANK1, DAGK, PI4Kβ, and CDPK1 across malaria species. DAGK, PANK1, and PI4Kβ possessed nonsynonymous mutations; surprisingly, the mutations only occurred in the AQr parasites. PANK1 acquired Asn394His, while DAGK contained K270R and K292R mutations. PI4Kβ had Asp366Asn, Ser1367Arg, Tyr1394Asn and Asp1423Asn. We show downregulation of PANK1, DAGK, and PI4Kβ in the trophozoites but upregulation at the schizonts stages in the AQr parasites. Conclusions The selective acquisition of the mutations and the differential gene expression in AQ-resistant parasites may signify proteins under AQ pressure. The role of the mutations in the resistant parasites and their impact on drug responses require investigations using reverse genetics techniques in malaria parasites.https://openresearchafrica.org/articles/5-28/v3Lumefantrine Piperaquine Amodiaquine Resistance Plasmodium berghei kinaseeng |
| spellingShingle | Peter Mwitari Francis Kimani Daniel Kiboi Brenda Muriithi Jean Chepngetich Kevin Thiong'o Beatrice Gachie Jeremiah Gathirwa Mercy Jepkorir Amodiaquine drug pressure selects nonsynonymous mutations in pantothenate kinase 1, diacylglycerol kinase, and phosphatidylinositol-4 kinase in Plasmodium berghei ANKA [version 3; peer review: 1 approved, 3 approved with reservations, 1 not approved] Open Research Africa Lumefantrine Piperaquine Amodiaquine Resistance Plasmodium berghei kinase eng |
| title | Amodiaquine drug pressure selects nonsynonymous mutations in pantothenate kinase 1, diacylglycerol kinase, and phosphatidylinositol-4 kinase in Plasmodium berghei ANKA [version 3; peer review: 1 approved, 3 approved with reservations, 1 not approved] |
| title_full | Amodiaquine drug pressure selects nonsynonymous mutations in pantothenate kinase 1, diacylglycerol kinase, and phosphatidylinositol-4 kinase in Plasmodium berghei ANKA [version 3; peer review: 1 approved, 3 approved with reservations, 1 not approved] |
| title_fullStr | Amodiaquine drug pressure selects nonsynonymous mutations in pantothenate kinase 1, diacylglycerol kinase, and phosphatidylinositol-4 kinase in Plasmodium berghei ANKA [version 3; peer review: 1 approved, 3 approved with reservations, 1 not approved] |
| title_full_unstemmed | Amodiaquine drug pressure selects nonsynonymous mutations in pantothenate kinase 1, diacylglycerol kinase, and phosphatidylinositol-4 kinase in Plasmodium berghei ANKA [version 3; peer review: 1 approved, 3 approved with reservations, 1 not approved] |
| title_short | Amodiaquine drug pressure selects nonsynonymous mutations in pantothenate kinase 1, diacylglycerol kinase, and phosphatidylinositol-4 kinase in Plasmodium berghei ANKA [version 3; peer review: 1 approved, 3 approved with reservations, 1 not approved] |
| title_sort | amodiaquine drug pressure selects nonsynonymous mutations in pantothenate kinase 1 diacylglycerol kinase and phosphatidylinositol 4 kinase in plasmodium berghei anka version 3 peer review 1 approved 3 approved with reservations 1 not approved |
| topic | Lumefantrine Piperaquine Amodiaquine Resistance Plasmodium berghei kinase eng |
| url | https://openresearchafrica.org/articles/5-28/v3 |
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