Identifying the Target of a Potent and Selective Inhibitor of Babesia Blood Stages.

确定巴贝虫血液阶段的有效和选择性抑制剂的靶标。

基本信息

批准号：
10661759
负责人：
Bjorn Felix Caesar Kafsack
金额：
$ 21.19万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
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：
2022-07-07 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10661759
关键词：
Annual Reports Babesia Babesiosis Behavior Binding Binding Proteins Biochemical Biological Assay Blood C10 CRISPR/Cas technology Candidate Disease Gene Case Study Cells Code Codon Nucleotides DNA Resequencing DNA methyltransferase inhibition Data Detection Development Drug Targeting Engineering Ensure Enzymes Escherichia coli Genes Genetic Genomics Human Immune system Immunocompromised Host Incidence Individual Infection Isomerism Label Life Methods Molecular Target Mutation Orthologous Gene Parasites Parents Predisposition Proteins Proteomics Protozoa Recombinants Reporting Resistance Scanning Sensitivity and Specificity Single Nucleotide Polymorphism Specificity System Testing Tick-Borne Diseases Toxic effect United States Validation Variant Vector-transmitted infectious disease Work Zoonoses analog candidate validation chemoproteomics design drug development experience experimental study genetic approach genome editing human DNA improved inhibitor knock-down nanomolar novel overexpression protein purification resistance mechanism sequencing platform side effect small molecule small molecule inhibitor tick transmission tick-borne tool vector whole genome

项目摘要

Project Summary Tick-borne diseases, including human babesiosis, are on the rise. Caused by protozoa in the genus Babesia, this infection can be life-threatening in asplenic or immunocompromised individuals. Based on the most recent CDC data, the number of annual reported cases in the US nearly doubled between 2011 and 2018, with some states experiencing a greater than 10-fold increase. Treatment options for babesiosis in immunocompromised individuals are very limited, have substantial side effects, and often fail to fully clear the parasites. This makes the development of new treatments imperative. We recently developed the small molecule inhibitor C10 that has low toxicity and excellent activity against multiple species of Babesia parasites that infect humans. The molecular target of this inhibitor is clearly druggable, essential, and likely novel but is currently unknown. Using a combination of genetic and biochemical approaches, the proposed experiments will identify and validate the parasite target of this inhibitor. In AIM1 we will identify what parasite proteins that bind tightly to C10 using µMap, a newly developed photocatalytic chemo-proteomics platform with greatly enhanced sensitivity and specificity over current methods. In a parallel effort, we have selected multiple independent parasite lines with partial resistance to C10. Since mutation in genes encoding a drug target is the most common resistance mechanism, AIM2 will use whole-genome resequencing to identify genes with newly acquired mutations in resistant lines but absent from the susceptible parent to identify an additional set of target candidates. Finally, we will validate target candidates from AIM1 and AIM2 by confirming that engineered mutations of the target, such as knockdown or overexpression, alters susceptibility to C10. If feasible, we will also test if C10 binds to and inhibits the activity of the recombinantly expressed target. Throughout these experiments will use C13, a nearly identical but inactive isomer of C10, to ensure target specificity.

项目摘要 tick传播的疾病，包括人类的bab虫病，正在上升。由原生动物在巴布西亚属中引起，这种感染可能会在浮力或免疫功能低下的个体中威胁生命。基于最近的 CDC数据，2011年至2018年之间，美国报告的年度报告数字几乎翻了一番州的增长超过10倍。免疫功能低下的Babesiosis的治疗选择个体非常有限，具有实质性的副作用，并且通常无法完全清除寄生虫。这使得必须开发新疗法。我们最近开发了具有的小分子抑制剂C10 低毒性和对感染人类的贝贝西亚寄生虫的卓越活性。分子该抑制剂的靶标显然是可吸毒，必不可少的，可能是新颖的，但目前尚不清楚。使用遗传和生化方法的结合，提出的实验将识别和验证该抑制剂的寄生虫靶标。在AIM1中，我们将确定哪些使用µMAP紧密结合与C10的寄生虫蛋白，一个新开发的光催化化学蛋白质学平台，具有良好的灵敏度和特异性超过当前方法。在平行的努力中，我们选择了多个具有部分独立的寄生虫线对C10的抵抗力。由于编码药物靶靶的基因中的突变是最常见的抗性机制，因此 AIM2将使用全基因组重新取样来鉴定具有抗性线新获得突变的基因，但易感父母没有确定一组额外的目标候选者。最后，我们将验证目标通过确认目标的工程突变，例如敲低或过表达，改变了对C10的敏感性。如果可行，我们还将测试C10是否与重组表达的目标。在这些实验中，将使用C13，这是一种几乎相同但不活跃的 C10的异构体，以确保目标特异性。