{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:59:25Z","timestamp":1785405565455,"version":"3.56.0"},"posted":{"date-parts":[[2026,7,7]]},"group-title":"Molecular Biology","reference-count":55,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T00:00:00Z","timestamp":1783382400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["AI127338"],"award-info":[{"award-number":["AI127338"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Malaria remains a pressing global health challenge, with the continued emergence of resistance threatening the long-term efficacy of artemisinin-based combination therapies (ACTs). Piperaquine (PPQ), an important partner drug in artemisinin-based combination therapies exhibits a unique bimodal dose-response phenotype associated with reduced susceptibility, yet the biological mechanism underlying this phenotype remains unknown. This phenotype is strongly associated with mutations in pfcrt and copy number amplification of\n                  <jats:italic>plasmepsin II\/III<\/jats:italic>\n                  (\n                  <jats:italic>pm II\/III<\/jats:italic>\n                  ). Given that plasmepsins play a central role in hemoglobin degradation within the blood stage parasite digestive vacuole, and that PPQ accumulates within this compartment and perturbs heme detoxification, this phenotype likely reflects alterations in fundamental biological processes alongside drug-specific effects. We used isogenic PPQ-resistant parasite clones differing only in\n                  <jats:italic>pm II\/III<\/jats:italic>\n                  copy number to integrate phenotypes with metabolic changes, and transcriptional responses to ascertain the impact of genotype combinations on parasite response to PPQ. Across increasing PPQ concentrations, parasites with elevated\n                  <jats:italic>pm II\/III<\/jats:italic>\n                  copy number exhibited distinct metabolic responses compared to single-copy parasites, specifically, an altered abundance of peptides derived from hemoglobin degradation, directly implicating a core biological pathway long associated with plasmepsin function. The combination of metabolic and transcriptional data with phenotypic measurements supports a model in which increased plasmepsin expression enhances the parasite\u2019s capacity to sustain hemoglobin digestion and associated metabolic activity under high PPQ concentrations. This points to a mechanistic basis for continued parasite survival, indicating that changes in hemoglobin processing within the digestive vacuole contribute to the bimodal response to PPQ. Molecular dynamics simulations further support a direct interaction between PPQ and PM II\/III, as a mechanism by which these proteins impact PPQ response dynamics through both modulation of hemoglobin digestion and protein-drug interactions within the digestive vacuole.\n                <\/jats:p>","DOI":"10.64898\/2026.07.06.734584","type":"posted-content","created":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T13:40:13Z","timestamp":1783431613000},"source":"Crossref","is-referenced-by-count":0,"title":["A Multidimensional Analysis of the Bimodal Piperaquine Response in\n                  <i>Plasmodium falciparum<\/i>"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4917-7205","authenticated-orcid":false,"given":"John","family":"Kane","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, Eck Institute for Global Health, University of Notre Dame, Notre Dame, Indiana, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aubrey","family":"Schall","sequence":"additional","affiliation":[{"name":"Dept. of Chemistry and Dept. of Biochemistry and Cellular and Molecular Biology, Georgetown University, Washington, D.C., USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa A.","family":"Checkley","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Eck Institute for Global Health, University of Notre Dame, Notre Dame, Indiana, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Douglas A.","family":"Shoue","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Eck Institute for Global Health, University of Notre Dame, Notre Dame, Indiana, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sydney M.","family":"Gavula","sequence":"additional","affiliation":[{"name":"Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania, USA"},{"name":"Huck Center for Malaria Research, The Pennsylvania State University, University Park, Pennsylvania, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caroline","family":"Thomas","sequence":"additional","affiliation":[{"name":"Dept. of Chemistry and Dept. of Biochemistry and Cellular and Molecular Biology, Georgetown University, Washington, D.C., USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xue","family":"Li","sequence":"additional","affiliation":[{"name":"Disease Intervention and Prevention Program, Texas Biomedical Research Institute, San Antonio, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian H.","family":"Cheeseman","sequence":"additional","affiliation":[{"name":"Host Pathogen Interactions Program, Texas Biomedical Research Institute, San Antonio, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashley M.","family":"Vaughan","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute, Seattle, Washington, USA"},{"name":"Department of Pediatrics, University of Washington, Seattle, Washington, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0191-0204","authenticated-orcid":false,"given":"Timothy J.","family":"Anderson","sequence":"additional","affiliation":[{"name":"Disease Intervention and Prevention Program, Texas Biomedical Research Institute, San Antonio, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manuel","family":"Llin\u00e1s","sequence":"additional","affiliation":[{"name":"Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania, USA"},{"name":"Huck Center for Malaria Research, The Pennsylvania State University, University Park, Pennsylvania, USA"},{"name":"Department of Biochemistry & Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul D.","family":"Roepe","sequence":"additional","affiliation":[{"name":"Dept. of Chemistry and Dept. of Biochemistry and Cellular and Molecular Biology, Georgetown University, Washington, D.C., USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael T.","family":"Ferdig","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Eck Institute for Global Health, University of Notre Dame, Notre Dame, Indiana, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026071003550724000_2026.07.06.734584v1.1","unstructured":"World Health Organization. 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