Mechanisms of Plasmodium vivax chloroquine resistance: a transcriptomic/transgenic approach

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. Themarkers and mechanism of chloroquine resistance (CQR) remain unknown; hampering molecularsurveillance and accurate diagnosis of CQR. Our hypothesis, based on our clinical study in Vietnam, published data from a non-human primate model and observations from Brazil, suggest that altered gene expression of transporter genes plays a major role in PvCQR. PvRESIST will capitalize on a collection of existing P. vivax clinical samples from Brazil with well-characterized CQ susceptibility to apply 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, will be generated in PvRESIST using advanced CRISPR-Cas9 genome editing strategies, in order to determine underlyingdrug resistance mechanism. Outcomes of PvRESIST heightens the research efforts so far and pushes boundaries of predescent research findings by for instance directly benefiting P. vivax patients and drug resistance surveillance, while advancing research with new tools, datasets and transgenic lines to investigate P. vivax biology.