{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T16:19:51Z","timestamp":1780589991968,"version":"3.54.1"},"reference-count":27,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,4,22]],"date-time":"2025-04-22T00:00:00Z","timestamp":1745280000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Building a New Generation of Academics (BANGA Africa, University of Ghana and Carnegie Corporation of New York)","award":["PID2021-128325OB-I00"],"award-info":[{"award-number":["PID2021-128325OB-I00"]}]},{"name":"Building a New Generation of Academics (BANGA Africa, University of Ghana and Carnegie Corporation of New York)","award":["MCIU\/AEI\/ 10.13039\/501100011033"],"award-info":[{"award-number":["MCIU\/AEI\/ 10.13039\/501100011033"]}]},{"name":"Building a New Generation of Academics (BANGA Africa, University of Ghana and Carnegie Corporation of New York)","award":["2021-SGR-00635"],"award-info":[{"award-number":["2021-SGR-00635"]}]},{"name":"WACCBIP-World Bank ACE PhD fellowship (WACCBIP+NCDs: Awandare)","award":["PID2021-128325OB-I00"],"award-info":[{"award-number":["PID2021-128325OB-I00"]}]},{"name":"WACCBIP-World Bank ACE PhD fellowship (WACCBIP+NCDs: Awandare)","award":["MCIU\/AEI\/ 10.13039\/501100011033"],"award-info":[{"award-number":["MCIU\/AEI\/ 10.13039\/501100011033"]}]},{"name":"WACCBIP-World Bank ACE PhD fellowship (WACCBIP+NCDs: Awandare)","award":["2021-SGR-00635"],"award-info":[{"award-number":["2021-SGR-00635"]}]},{"name":"Ministerio de Ciencia, Innovaci\u00f3n y Universidades\/Agencia Estatal de Investigaci\u00f3n","award":["PID2021-128325OB-I00"],"award-info":[{"award-number":["PID2021-128325OB-I00"]}]},{"name":"Ministerio de Ciencia, Innovaci\u00f3n y Universidades\/Agencia Estatal de Investigaci\u00f3n","award":["MCIU\/AEI\/ 10.13039\/501100011033"],"award-info":[{"award-number":["MCIU\/AEI\/ 10.13039\/501100011033"]}]},{"name":"Ministerio de Ciencia, Innovaci\u00f3n y Universidades\/Agencia Estatal de Investigaci\u00f3n","award":["2021-SGR-00635"],"award-info":[{"award-number":["2021-SGR-00635"]}]},{"name":"FEDER and European Union NextGenerationEU\/PRTR","award":["PID2021-128325OB-I00"],"award-info":[{"award-number":["PID2021-128325OB-I00"]}]},{"name":"FEDER and European Union NextGenerationEU\/PRTR","award":["MCIU\/AEI\/ 10.13039\/501100011033"],"award-info":[{"award-number":["MCIU\/AEI\/ 10.13039\/501100011033"]}]},{"name":"FEDER and European Union NextGenerationEU\/PRTR","award":["2021-SGR-00635"],"award-info":[{"award-number":["2021-SGR-00635"]}]},{"name":"Generalitat de Catalunya","award":["PID2021-128325OB-I00"],"award-info":[{"award-number":["PID2021-128325OB-I00"]}]},{"name":"Generalitat de Catalunya","award":["MCIU\/AEI\/ 10.13039\/501100011033"],"award-info":[{"award-number":["MCIU\/AEI\/ 10.13039\/501100011033"]}]},{"name":"Generalitat de Catalunya","award":["2021-SGR-00635"],"award-info":[{"award-number":["2021-SGR-00635"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Our understanding of the molecular mechanisms undergirding artemisinin (ART) resistance in Plasmodium falciparum is currently based on two organizing principles: reduced hemoglobin trafficking into the digestive food vacuole, resulting in lower levels of activated ART, and increased tolerance to ART-induced oxidative stress in the infected erythrocyte. We had previously proposed an extracellular vesicle (EV) export model of ART resistance in P. falciparum. This model predicts that EV abundance will be altered by ART exposure and that the peptide cargo of EVs from the ART-exposed condition will be enriched with aggregation-prone peptides. We tested the predictions of the EV export hypothesis in this study using in vitro culture assays of an ART-resistant transgenic line engineered on a 3D7 background (R561H) and a 3D7 knock-out line (PfVps60KO) with deficient EV production phenotype. EV enrichment was obtained from in vitro parasite culture supernatants via a series of ultracentrifugation and filtration steps, followed by size exclusion chromatography. A quality check on EVs was performed using dynamic light scattering. Liquid chromatography with tandem mass spectrometry was used to determine the proteome cargo from extracted EVs, and parasite peptides were queried for aggregation-prone tendency using open-access software. We report that dihydroartemisinin (DHA) exposure was positively correlated with EV abundance (coefficient estimate = 1038.58, confidence interval of 194.86\u20131882.30, and p-value = 0.018) and suggests that EV biogenesis is part of the parasite\u2019s response to DHA\/ART. Furthermore, our findings suggest the expression of a non-constitutive DHA-induced alternate EV biogenesis pathway as the PfVps60KO was observed to produce the highest number of EVs under DHA exposure. Finally, we show that EVs from both ART-susceptible and resistant parasites under DHA exposure carry a cargo of Chorein N-terminal domain-containing protein (PF3D7_1021700) with a high aggregation-prone index (prion-like domain [PrLD] score = 26.5) out of nine identified parasite peptides. The former of these findings is in concordance with the EV export hypothesis, which posits that the removal of DHA\/ART-induced aggregated and\/or misfolded peptides is critical to the parasite\u2019s survival under DHA\/ART exposure. This observation further implicates EVs in the development of the ART-resistant phenotype. However, the finding of one aggregation-prone peptide out of the nine parasite proteins in the EV cargo does not sufficiently support the EV export hypothesis. Future replicates of this study and further interrogations of the EV export hypothesis are needed.<\/jats:p>","DOI":"10.3390\/ijms26093962","type":"journal-article","created":{"date-parts":[[2025,4,22]],"date-time":"2025-04-22T20:24:52Z","timestamp":1745353492000},"page":"3962","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Extracellular Vesicle Abundance, but Not a High Aggregation-Prone Peptide Cargo, Is Associated with Dihydroartemisinin Exposure in Plasmodium falciparum"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1628-2845","authenticated-orcid":false,"given":"Kwesi Z.","family":"Tandoh","sequence":"first","affiliation":[{"name":"West African Centre for Cell Biology of Infectious Pathogens, Department of Biochemistry, Cell and Molecular Biology, College of Basic and Applied Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"},{"name":"Department of Epidemiology, Noguchi Memorial Institute for Medical Research, College of Health Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4074-5624","authenticated-orcid":false,"given":"Yunuen","family":"Avalos-Padilla","sequence":"additional","affiliation":[{"name":"Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), Baldiri Reixac 10-12, 08028 Barcelona, Spain"},{"name":"Barcelona Institute for Global Health (ISGlobal, Hospital Cl\u0131nic-Universitat de Barcelona), Rossell\u00f3 149-153, 08036 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Prince","family":"Ameyaw","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, Noguchi Memorial Institute for Medical Research, College of Health Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisabeth K.","family":"Laryea-Akrong","sequence":"additional","affiliation":[{"name":"West African Centre for Cell Biology of Infectious Pathogens, Department of Biochemistry, Cell and Molecular Biology, College of Basic and Applied Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"},{"name":"Department of Epidemiology, Noguchi Memorial Institute for Medical Research, College of Health Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gordon A.","family":"Awandare","sequence":"additional","affiliation":[{"name":"West African Centre for Cell Biology of Infectious Pathogens, Department of Biochemistry, Cell and Molecular Biology, College of Basic and Applied Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6048-8509","authenticated-orcid":false,"given":"Michael David","family":"Wilson","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Noguchi Memorial Institute for Medical Research, College of Health Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Neils B.","family":"Quashie","sequence":"additional","affiliation":[{"name":"West African Centre for Cell Biology of Infectious Pathogens, Department of Biochemistry, Cell and Molecular Biology, College of Basic and Applied Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"},{"name":"Department of Epidemiology, Noguchi Memorial Institute for Medical Research, College of Health Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"},{"name":"Centre for Tropical Clinical Pharmacology and Therapeutics, School of Medicine and Dentistry, College of Health Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4622-9631","authenticated-orcid":false,"given":"Xavier","family":"Fern\u00e0ndez-Busquets","sequence":"additional","affiliation":[{"name":"Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), Baldiri Reixac 10-12, 08028 Barcelona, Spain"},{"name":"Barcelona Institute for Global Health (ISGlobal, Hospital Cl\u0131nic-Universitat de Barcelona), Rossell\u00f3 149-153, 08036 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8819-1793","authenticated-orcid":false,"given":"Nancy O.","family":"Duah-Quashie","sequence":"additional","affiliation":[{"name":"West African Centre for Cell Biology of Infectious Pathogens, Department of Biochemistry, Cell and Molecular Biology, College of Basic and Applied Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"},{"name":"Department of Epidemiology, Noguchi Memorial Institute for Medical Research, College of Health Sciences, University of Ghana, Legon, Accra P.O. Box LG 54, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1056\/NEJMoa0808859","article-title":"Artemisinin resistance in Plasmodium falciparum malaria","volume":"361","author":"Dondorp","year":"2009","journal-title":"N. Engl. J. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2619","DOI":"10.1056\/NEJMc0805011","article-title":"Evidence of artemisinin-resistant malaria in western Cambodia","volume":"359","author":"Noedl","year":"2008","journal-title":"N. Engl. J. Med."},{"key":"ref_3","unstructured":"WHO (2024, November 26). World Malaria Report 2023. 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