Artemisinin resistance phenotypes and K13 inheritance in a Plasmodium falciparum cross and Aotus model.
Sá JM., Kaslow SR., Krause MA., Melendez-Muniz VA., Salzman RE., Kite WA., Zhang M., Moraes Barros RR., Mu J., Han PK., Mershon JP., Figan CE., Caleon RL., Rahman RS., Gibson TJ., Amaratunga C., Nishiguchi EP., Breglio KF., Engels TM., Velmurugan S., Ricklefs S., Straimer J., Gnädig NF., Deng B., Liu A., Diouf A., Miura K., Tullo GS., Eastman RT., Chakravarty S., James ER., Udenze K., Li S., Sturdevant DE., Gwadz RW., Porcella SF., Long CA., Fidock DA., Thomas ML., Fay MP., Sim BKL., Hoffman SL., Adams JH., Fairhurst RM., Su X-Z., Wellems TE.
Concerns about malaria parasite resistance to treatment with artemisinin drugs (ARTs) have grown with findings of prolonged parasite clearance t 1/2s (>5 h) and their association with mutations in Plasmodium falciparum Kelch-propeller protein K13. Here, we describe a P. falciparum laboratory cross of K13 C580Y mutant with C580 wild-type parasites to investigate ART response phenotypes in vitro and in vivo. After genotyping >400 isolated progeny, we evaluated 20 recombinants in vitro: IC50 measurements of dihydroartemisinin were at similar low nanomolar levels for C580Y- and C580-type progeny (mean ratio, 1.00; 95% CI, 0.62-1.61), whereas, in a ring-stage survival assay, the C580Y-type progeny had 19.6-fold (95% CI, 9.76-39.2) higher average counts. In splenectomized Aotus monkeys treated with three daily doses of i.v. artesunate, t 1/2 calculations by three different methods yielded mean differences of 0.01 h (95% CI, -3.66 to 3.67), 0.80 h (95% CI, -0.92 to 2.53), and 2.07 h (95% CI, 0.77-3.36) between C580Y and C580 infections. Incidences of recrudescence were 57% in C580Y (4 of 7) versus 70% in C580 (7 of 10) infections (-13% difference; 95% CI, -58% to 35%). Allelic substitution of C580 in a C580Y-containing progeny clone (76H10) yielded a transformant (76H10C580Rev) that, in an infected monkey, recrudesced regularly 13 times over 500 d. Frequent recrudescences of ART-treated P. falciparum infections occur with or without K13 mutations and emphasize the need for improved partner drugs to effectively eliminate the parasites that persist through the ART component of combination therapy.