TY - JOUR
T1 - Generation of a mutator parasite to drive resistome discovery in Plasmodium falciparum
AU - Kümpornsin, Krittikorn
AU - Kochakarn, Theerarat
AU - Yeo, Tomas
AU - Okombo, John
AU - Luth, Madeline R.
AU - Hoshizaki, Johanna
AU - Rawat, Mukul
AU - Pearson, Richard D.
AU - Schindler, Kyra A.
AU - Mok, Sachel
AU - Park, Heekuk
AU - Uhlemann, Anne-Catrin
AU - Jana, Gouranga P.
AU - Maity, Bikash C.
AU - Laleu, Benoît
AU - Chenu, Elodie
AU - Duffy, James
AU - Moliner Cubel, Sonia
AU - Franco, Virginia
AU - Gomez-Lorenzo, Maria G.
AU - Gamo, Francisco Javier
AU - Winzeler, Elizabeth A.
AU - Fidock, David A.
AU - Chookajorn, Thanat
AU - Lee, Marcus C. S.
N1 - Funding Infomation:
We would like to acknowledge funding from the Bill and Melinda Gates Foundation to M.C.S.L., D.A.F., G.S.K., and E.A.W. (OPP1054480), and to D.A.F. (INV-033538), funding from the Wellcome Institutional Translational Partnership award (ITPA) and Mahidol University to T.K. and T.C., and funding from Wellcome [206194/Z/17/Z] to M.C.S.L. and the Wellcome Sanger Institute.
Copyright:
© 2023. The Author(s).
PY - 2023/5/27
Y1 - 2023/5/27
N2 - In vitro evolution of drug resistance is a powerful approach for identifying antimalarial targets, however, key obstacles to eliciting resistance are the parasite inoculum size and mutation rate. Here we sought to increase parasite genetic diversity to potentiate resistance selections by editing catalytic residues of Plasmodium falciparum DNA polymerase δ. Mutation accumulation assays reveal a ~5-8 fold elevation in the mutation rate, with an increase of 13-28 fold in drug-pressured lines. Upon challenge with the spiroindolone PfATP4-inhibitor KAE609, high-level resistance is obtained more rapidly and at lower inocula than wild-type parasites. Selections also yield mutants with resistance to an "irresistible" compound, MMV665794 that failed to yield resistance with other strains. We validate mutations in a previously uncharacterised gene, PF3D7_1359900, which we term quinoxaline resistance protein (QRP1), as causal for resistance to MMV665794 and a panel of quinoxaline analogues. The increased genetic repertoire available to this "mutator" parasite can be leveraged to drive P. falciparum resistome discovery.
AB - In vitro evolution of drug resistance is a powerful approach for identifying antimalarial targets, however, key obstacles to eliciting resistance are the parasite inoculum size and mutation rate. Here we sought to increase parasite genetic diversity to potentiate resistance selections by editing catalytic residues of Plasmodium falciparum DNA polymerase δ. Mutation accumulation assays reveal a ~5-8 fold elevation in the mutation rate, with an increase of 13-28 fold in drug-pressured lines. Upon challenge with the spiroindolone PfATP4-inhibitor KAE609, high-level resistance is obtained more rapidly and at lower inocula than wild-type parasites. Selections also yield mutants with resistance to an "irresistible" compound, MMV665794 that failed to yield resistance with other strains. We validate mutations in a previously uncharacterised gene, PF3D7_1359900, which we term quinoxaline resistance protein (QRP1), as causal for resistance to MMV665794 and a panel of quinoxaline analogues. The increased genetic repertoire available to this "mutator" parasite can be leveraged to drive P. falciparum resistome discovery.
KW - Animals
KW - Plasmodium falciparum/genetics
KW - Parasites/metabolism
KW - Malaria, Falciparum/drug therapy
KW - Antimalarials/therapeutic use
KW - Mutation
KW - Drug Resistance/genetics
KW - Protozoan Proteins/metabolism
UR - http://www.scopus.com/inward/record.url?scp=85160271952&partnerID=8YFLogxK
U2 - 10.1038/s41467-023-38774-1
DO - 10.1038/s41467-023-38774-1
M3 - Article
C2 - 37244916
SN - 2041-1723
VL - 14
JO - Nature Communications
JF - Nature Communications
M1 - 3059
ER -
