Evaluating host-directed therapeutics against blood-stage malaria parasites

总结 疟疾仍然是一个重大的全球健康问题，造成巨大的发病率和死亡率。疾病发生 在人类感染的血液阶段，红细胞中疟原虫寄生虫的无性繁殖 （红细胞）导致其破坏。由于耐药性的威胁不断演变，迫切需要 开发新的抗疟药物，以控制和根除疟疾。我们假设宿主定向抑制剂（HDIs） 将绕过寄生虫的抗药性，这将不得不显着改变其生物学，以绕过其 基本主机目标的要求。越来越多的研究报道了RBC特异性抑制剂， 但这些都没有得到很好的验证。我们建议识别和验证HDIs。我们将包括鉴定的化合物 通过在RBC蛋白质组中筛选具有推定靶点的大型药物再利用文库。验证主机 靶向作为抗疟作用机制，我们将使用体外培养的 具有推定靶标敲低的RBC（cRBC）。细胞热位移试验将用于证明 直接与宿主靶点结合。最后，对于看起来是真正的HDIs的化合物，我们将评估其 产生抗药性。建立一个管道，以验证作为机制的东道国目标 假定的HDIs的作用将是该领域的一个重大进展，支持宿主导向的 未来的治疗。

Summary Malaria remains a major global health problem, causing tremendous morbidity and mortality. Disease occurs during the blood-stage of human infections with asexual proliferation of Plasmodium parasites in red blood cells (RBCs) leading to their destruction. Due to the evolving threat of drug resistance, there is an urgent need to develop new antimalarials for control and eradication efforts. We hypothesize that host-directed inhibitors (HDIs) will circumvent drug resistance in the parasite, which would have to significantly alter its biology to bypass its requirement for the essential host target. An increasing number of studies have reported RBC-specific inhibitors, but these are poorly validated. We propose to identify and validate HDIs. We will include compounds identified by screening a large drug repurposing library with putative targets in the RBC proteome. To validate host targeting as the antimalarial mechanism of action, we will perform synthetic lethal assays using in vitro cultured RBCs (cRBCs) with knockdown of putative targets. Cellular Thermal Shift Assays will be used to demonstrate direct binding to host targets. Finally, for compounds that appear to be bona fide HDIs, we will assess the ability of the parasite to develop resistance. The establishment of a pipeline to validate host targeting as the mechanism of action of putative HDIs will be a significant advance to the field, supporting the development of host-directed therapeutics in the future.