Mechanisms of P. vivax chloroquine resistance: a transcriptomic/transgenic approach
Investigate the role of gene expression in PvCQR parasites from clinical samples using innovative transcriptomic approaches follewed by a transgenic P. knowlesi model to provide direct evidence for the mechanism of resistance.
Plasmodium vivax is the most predominant species outside Africa, and many countries aiming towards malaria elimination by 2030 are seeing an increase in resistance against chloroquine. The markers and mechanism of chloroquine resistance (CQR) remain unknown; hampering molecular surveillance and accurate diagnosis of CQR. Our hypothesis, based on our results from a clinical study in Vietnam and recent published data obtained from parasites adapted to a non-human primate model, suggest that altered gene expression of transporter genes plays a major role in PvCQR. We will capitalize on a large collection of existing P. vivax clinical samples (also with CQR) at the Malariology Unit to pioneer in the application of cutting-edge RNA sequencing technologies, including single-cell transcriptomics. We will unravel the transcriptional network of genes underlying PvCQR, and the impact of infection complexity (parasite life-stages and mixed clones present in natural infections) in treatment outcome. P. knowlesi transgenic lines, differentially expressing P. vivax genes, are generated using advanced CRISPR-Cas9 genome editing strategies, in order to determine underlying drug resistance mechanism. Outcomes of the study will directly benefit P. vivax patients and drug resistance surveillance, while advancing research with new protocols, tools, datasets and transgenic lines to investigate P. vivax biology.