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

• 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
• 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

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.

摘要 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。疾病会发生 人感染血液期红细胞内疟原虫无性繁殖 (RBCs)导致它们的毁灭。由于不断变化的抗药性威胁，迫切需要 为控制和根除努力开发新的抗疟疾药物。我们假设宿主导向的抑制物(HDI) 将绕过寄生虫的抗药性，这将不得不显著改变其生物学特性以绕过其 对基本主机目标的要求。越来越多的研究报告了RBC特异性抑制剂， 但这些都没有得到充分的验证。我们建议识别和验证人类发展指数。我们将包括已确定的化合物 通过在RBC蛋白质组中筛选具有假定靶点的大型药物再利用文库。验证主机 以抗疟疾的作用机制为目标，我们将使用体外培养的 击倒假定目标的RBC(CRBCs)。细胞热位移分析将用于演示 直接绑定到主机目标。最后，对于看起来是真正的HDI的化合物，我们将评估其能力 以产生抵抗力。建立管道以验证主机目标作为机制 假定的人类发展倡议行动的实施将是对该领域的重大进步，支持以主机为导向的发展 未来的治疗学。