Evaluating host-directed therapeutics against blood-stage malaria parasites

评估针对血期疟疾寄生虫的宿主导向疗法

基本信息

批准号：
10665779
负责人：
Manoj T Duraisingh
金额：
$ 23.93万
依托单位：
HARVARD SCHOOL OF PUBLIC HEALTH
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-14 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10665779
关键词：
Acceleration Antimalarials Binding Biochemical Biological Assay Biology Blood Bypass CD34 gene CRISPR/Cas technology Cell Culture System Cell Cycle Cell Differentiation process Cell Nucleus Cells Clinical Clinical Trials Development Disease Drug Kinetics Drug Screening Drug resistance Erythrocytes Future Generations Genes Genetic Growth HIV Hematopoietic stem cells Hepatitis Human In Vitro Infection Integration Host Factors Lentivirus Libraries Literature Malaria Methods Molecular Target Morbidity - disease rate Parasites Pharmaceutical Preparations Plasmodium Plasmodium falciparum Podophyllotoxin Positioning Attribute Proliferating Proteins Proteome RNA Interference Reporting Resistance Resistance development Safety Specificity Therapeutic Toxic effect Validation Virus asexual chemical genetics cost drug repurposing genetic manipulation global health in vitro Assay indexing inhibitor knock-down mortality mutant next generation operation prevent screening small molecule inhibitor success therapeutic development

项目摘要

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.

概括疟疾仍然是一个主要的全球健康问题，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，疟原虫寄生虫在红细胞中无性增殖（红细胞）导致其破坏。由于耐药性威胁不断演变，迫切需要开发新的抗疟药以进行控制和根除工作。我们假设宿主定向抑制剂（HDI）将规避寄生虫的耐药性，而寄生虫必须显着改变其生物学才能绕过其耐药性基本宿主目标的要求。越来越多的研究报道了红细胞特异性抑制剂，但这些都没有得到很好的验证。我们建议识别和验证 HDI。我们将包括已识别的化合物通过筛选具有红细胞蛋白质组中推定目标的大型药物再利用库。验证主机作为抗疟作用机制的目标，我们将使用体外培养的合成致死试验敲低假定目标的红细胞 (cRBC)。细胞热位移测定将用于证明直接结合到宿主目标。最后，对于看似真正的 HDI 的化合物，我们将评估其能力寄生虫产生抗药性。建立一个管道来验证主机定位作为机制假定的人类发展倡议的行动将是该领域的重大进步，支持东道主导向的发展未来的治疗方法。