{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T14:47:59Z","timestamp":1785422879179,"version":"3.56.0"},"posted":{"date-parts":[[2026,4,3]]},"group-title":"Microbiology","reference-count":67,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T00:00:00Z","timestamp":1775174400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004167","name":"Medicines for Malaria Venture","doi-asserted-by":"crossref","award":["RD-08-0015"],"award-info":[{"award-number":["RD-08-0015"]}],"id":[{"id":"10.13039\/501100004167","id-type":"DOI","asserted-by":"crossref"}]},{"award":["LT000976\/2016-L"],"award-info":[{"award-number":["LT000976\/2016-L"]}],"id":[{"id":"https:\/\/ror.org\/02ebx7v45","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01 AI182318"],"award-info":[{"award-number":["R01 AI182318"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01 AI109023"],"award-info":[{"award-number":["R01 AI109023"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000865","name":"Bill & Melinda Gates Foundation","doi-asserted-by":"crossref","award":["OPP1054480"],"award-info":[{"award-number":["OPP1054480"]}],"id":[{"id":"10.13039\/100000865","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,4,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  New antimalarial compounds are urgently required to overcome artemisinin partial resistance that has emerged in Asia and now Africa. Ozonides are promising next-generation artemisinins that offer the improved pharmacokinetic property of a prolonged\n                  <jats:italic>in vivo<\/jats:italic>\n                  half-life. To assess the potential for parasite resistance to ozonides in an artemisinin-resistant background, we subjected Cambodian Kelch13 (K13) mutant parasites to increasing artefenomel (OZ439) pressure up to\n                  <jats:italic>in vivo<\/jats:italic>\n                  physiological concentrations. Whole-genome sequencing identified a novel non-propeller K13 A212T mutation in OZ439-resistant parasites. Gene editing and drug susceptibility assays revealed that the K13 double mutation R539T+A212T is a determinant of OZ439 resistance. In extended parasite recovery assays, this resistance mechanism was associated with accelerated parasite recrudescence following OZ439 or OZ277 exposure. This phenotype was also observed in K13 C580Y+A212T double mutant parasites. Global metabolomic profiling revealed no changes in the levels of hemoglobin-derived peptides in OZ439-resistant parasites, suggesting that resistance is not associated with drug activation. Instead, double mutant parasites exhibited increased levels of metabolites linked to glutathione, nucleotide, and aspartate-glutamate metabolism, suggesting a higher capacity for redox regulation to tolerate drug-induced oxidative damage. Our findings demonstrate that ozonide resistance can emerge through a novel K13 mutation on the background of existing artemisinin-resistance\n                  <jats:italic>k13<\/jats:italic>\n                  alleles.\n                <\/jats:p>","DOI":"10.64898\/2026.04.02.716214","type":"posted-content","created":{"date-parts":[[2026,4,4]],"date-time":"2026-04-04T03:00:14Z","timestamp":1775271614000},"source":"Crossref","is-referenced-by-count":0,"title":["Double Mutations in\n                  <i>Plasmodium falciparum<\/i>\n                  Kelch13 drive resistance to next-generation artemisinin derivatives in malaria parasites"],"prefix":"10.64898","author":[{"given":"Christopher","family":"Bower-Lepts","sequence":"first","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, Columbia University Irving Medical Center, New York, NY, USA"},{"name":"Center for Malaria Therapeutic and Antimicrobial Resistance, Columbia University Irving Medical Center, New 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USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomas","family":"Yeo","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Columbia University Irving Medical Center, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tarrick","family":"Qahash","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer L.","family":"Small-Saunders","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, Columbia University Irving Medical Center, New York, NY, USA"},{"name":"Center for Malaria Therapeutic and Antimicrobial Resistance, Columbia University Irving Medical Center, New York, NY, USA"},{"name":"Department of Microbiology and Immunology, Columbia University Irving Medical Center, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heekuk","family":"Park","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, Columbia University Irving Medical Center, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne-Catrin","family":"Uhlemann","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, Columbia University Irving Medical Center, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6173-5882","authenticated-orcid":false,"given":"Manuel","family":"Llin\u00e1s","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, PA, USA"},{"name":"Huck Center for Malaria Research, Pennsylvania State University, University Park, PA, USA"},{"name":"Department of Chemistry, Pennsylvania State University, University Park, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David A.","family":"Fidock","sequence":"additional","affiliation":[{"name":"Center for Malaria Therapeutic and Antimicrobial Resistance, Columbia University Irving Medical Center, New York, NY, USA"},{"name":"Department of Microbiology and Immunology, Columbia University Irving Medical Center, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sachel","family":"Mok","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, Columbia University Irving Medical Center, New York, NY, USA"},{"name":"Center for Malaria Therapeutic and Antimicrobial Resistance, Columbia University Irving Medical Center, New York, NY, USA"},{"name":"Aaron Diamond AIDS Research Center (ADARC), Columbia University Irving Medical Center, New York, NY, 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