title: A molecular mechanism of artemisinin resistance in Plasmodium falciparum malaria.
abstract: Artemisinins are the cornerstone of anti-malarial drugs. Emergence and spread of resistance to them raises risk of wiping out recent gains achieved in reducing worldwide malaria burden and threatens future malaria control and elimination on a global level. Genome-wide association studies (GWAS) have revealed parasite genetic loci associated with artemisinin resistance. However, there is no consensus on biochemical targets of artemisinin. Whether and how these targets interact with genes identified by GWAS, remains unknown. Here we provide biochemical and cellular evidence that artemisinins are potent inhibitors of Plasmodium falciparum phosphatidylinositol-3-kinase (PfPI3K), revealing an unexpected mechanism of action. In resistant clinical strains, increased PfPI3K was associated with the C580Y mutation in P. falciparum Kelch13 (PfKelch13), a primary marker of artemisinin resistance. Polyubiquitination of PfPI3K and its binding to PfKelch13 were reduced by the PfKelch13 mutation, which limited proteolysis of PfPI3K and thus increased levels of the kinase, as well as its lipid product phosphatidylinositol-3-phosphate (PI3P). We find PI3P levels to be predictive of artemisinin resistance in both clinical and engineered laboratory parasites as well as across non-isogenic strains. Elevated PI3P induced artemisinin resistance in absence of PfKelch13 mutations, but remained responsive to regulation by PfKelch13. Evidence is presented for PI3P-dependent signalling in which transgenic expression of an additional kinase confers resistance. Together these data present PI3P as the key mediator of artemisinin resistance and the sole PfPI3K as an important target for malaria elimination.
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors: Alassane
authors: A
authors: Mbengue
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors: Souvik
authors: S
authors: Bhattacharjee
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors: Trupti
authors: T
authors: Pandharkar
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors: Haining
authors: H
authors: Liu
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors: Guillermina
authors: G
authors: Estiu
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, USA [3] Department of Biochemistry &Molecular Biology, Indiana University School of Medicine-South Bend, 143 Raclin-Carmichael Hall, 1234 Notre Dame Avenue, South Bend, Indiana 46617, USA.
authors: Robert V
authors: RV
authors: Stahelin
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors: Shahir S
authors: SS
authors: Rizk
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA [3] Departmen of. Biochemistry and Molecular Biology, Faculty of Science University of Buea, P.O. Box 63 Buea, Southwest region, Cameroon.
authors: Dieudonne L
authors: DL
authors: Njimoh
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors: Yana
authors: Y
authors: Ryan
authors: Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Ratchathewi, Bangkok 10400, Thailand.
authors: Kesinee
authors: K
authors: Chotivanich
authors: National Center for Parasitology, Entomology and Malaria Control, 12302 Phnom Penh, Monivong Blvd, Phnom Penh 12302, Cambodia.
authors: Chea
authors: C
authors: Nguon
authors: CNRS 5290/IRD 224/University Montpellier 1&2 ("MiVEGEC"), Montpellier, France.
authors: Mehdi
authors: M
authors: Ghorbal
authors: CNRS 5290/IRD 224/University Montpellier 1&2 ("MiVEGEC"), Montpellier, France.
authors: Jose-Juan
authors: JJ
authors: Lopez-Rubio
authors: Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors: Michael
authors: M
authors: Pfrender
authors: Department of Computer Science and Engineering, University of Notre Dame, Notre Dame, Indiana 46556. USA.
authors: Scott
authors: S
authors: Emrich
authors: New York Blood Center, New York, New York 10032, USA.
authors: Narla
authors: N
authors: Mohandas
authors: 1] Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Ratchathewi, Bangkok 10400, Thailand [2] Centre for Tropical Medicine, Nuffield Department of Clinical Medicine, University of Oxford, Oxford OX3 7BN. UK.
authors: Arjen M
authors: AM
authors: Dondorp
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, USA [3] Laboratory of Computational Chemistry and Drug Design, Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen 518055, China.
authors: Olaf
authors: O
authors: Wiest
authors: 1] Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, Indiana 46556, USA [2] Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors: Kasturi
authors: K
authors: Haldar
mesh_terms: D000962:Antimalarials / Q000494:pharmacology*; D037621:Artemisinins / Q000494:pharmacology*; D004351:Drug Resistance / Q000187:drug effects* / Q000235:genetics; D055106:Genome-Wide Association Study; D016778:Malaria, Falciparum / Q000188:drug therapy* / Q000469:parasitology*; D008958:Models, Molecular; D009154:Mutation; D058539:Phosphatidylinositol 3-Kinase / Q000737:chemistry / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000201:enzymology*; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D000962:Antimalarials; D037621:Artemisinins; D018129:Phosphatidylinositol Phosphates; D000081082:Phosphoinositide-3 Kinase Inhibitors; D015800:Protozoan Proteins; C055525:phosphatidylinositol 3-phosphate; C039060:artenimol; C031327:artemisinin; D058539:Phosphatidylinositol 3-Kinase