title: Altered temporal response of malaria parasites determines differential sensitivity to artemisinin.
abstract: Reports of emerging resistance to first-line artemisinin antimalarials make it critical to define resistance mechanisms and identify in vitro correlates of resistance. Here we combine unique in vitro experimental and analytical approaches to mimic in vivo drug exposure in an effort to provide insight into mechanisms of drug resistance. Tightly synchronized parasites exposed to short drug pulses exhibit large stage-dependent differences in their drug response that correlate with hemoglobin digestion throughout most of the asexual cycle. As a result, ring-stage parasites can exhibit >100-fold lower sensitivity to short drug pulses than trophozoites, although we identify a subpopulation of rings (2-4 h postinvasion) that exhibits hypersensitivity. We find that laboratory strains that show little differences in drug sensitivity in standard in vitro assays exhibit substantial (>95-fold) difference in sensitivity when exposed to short drug pulses. These stage- and strain-dependent differences in drug sensitivity reflect differential response lag times with rings exhibiting lag times of up to 4 h. A simple model that assumes that the parasite experiences a saturable effective drug dose describes the complex dependence of parasite viability on both drug concentration and exposure time and is used to demonstrate that small changes in the parasite's drug response profile can dramatically alter the sensitivity to artemisinins. This work demonstrates that effective resistance can arise from the interplay between the short in vivo half-life of the drug and the stage-specific lag time and provides the framework for understanding the mechanisms of drug action and parasite resistance.
authors: Department of Biochemistry and Molecular Biology and Australian Research Council Centre of Excellence for Coherent X-Ray Science, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria 3010, Australia.
authors: Nectarios
authors: N
authors: Klonis
authors: Stanley C
authors: SC
authors: Xie
authors: James M
authors: JM
authors: McCaw
authors: Maria P
authors: MP
authors: Crespo-Ortiz
authors: Sophie G
authors: SG
authors: Zaloumis
authors: Julie A
authors: JA
authors: Simpson
authors: Leann
authors: L
authors: Tilley
mesh_terms: D037621:Artemisinins / Q000494:pharmacology*; D004351:Drug Resistance / Q000187:drug effects* / Q000502:physiology; D007783:Lactones / Q000494:pharmacology*; D008954:Models, Biological; D010963:Plasmodium falciparum / Q000166:cytology / Q000378:metabolism*; D013045:Species Specificity
chemical_list: D037621:Artemisinins; D007783:Lactones; C075875:artemisin