Integrating Drosophila and human podocyte studies to discover APOL1 renal toxicity mechanism and therapeutic targets

整合果蝇和人类足细胞研究发现APOL1肾毒性机制和治疗靶点

基本信息

批准号：
10319177
负责人：
ZHE HAN
金额：
$ 34.76万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10319177
关键词：
APOL1 gene African American population African ancestry Albuminuria Alleles Animal Model Autophagocytosis Azotemia Candidate Disease Gene Cell Culture Techniques Cells Data Defect Disease Down-Regulation Drosophila genus Drug Compounding Drug Screening Exhibits Face Foot Process Gene Expression Gene Expression Alteration Gene Expression Profile Genes Genetic Genetic Screening Human Insecta Kidney Kidney Diseases Mediating Medicine Modeling Molecular Mus Nature Nephrotic Syndrome Outcome Overlapping Genes Patients Persons Pharmaceutical Preparations Pharmacotherapy Publishing Renal glomerular disease Risk Role Screening procedure Series Study models System Testing Tissues Toxic effect Up-Regulation Validation Vesicle Work Yeasts base cell injury cost drug candidate drug use screening fly gene therapy genetic testing genome-wide glomerulosclerosis in vivo evaluation insight kidney cell mouse model mutant novel podocyte protective effect risk variant screening therapeutic target tool trafficking transcriptome sequencing

项目摘要

African Americans face elevated risk of kidney disease, due in large part to inheritance of APOL1 risk alleles (RA). The molecular and cellular mechanisms underlying APOL1 nephropathy are being extensively investigated, but the mechanism remains unclear and potential therapeutic targets remain hidden. We generated a Drosophila model to study APOL1 nephropathy and showed that APOL1-RA led to renal cell injury in nephrocytes, the insect structural and functional homologs of human podocytes. The recently developed mouse APOL1 model provides a critical tool to study the molecular mechanism of APOL1 nephropathy and to test promising treatments, but it cannot be used to conduct large-scale genetic screens to identify modifier genes that could antagonize the renal toxicity of APOL1-RA. It is also extremely difficult to use the mouse model to screen for hundreds of candidate drug compounds. The Drosophila system presents unique advantages in terms of low-cost and high-efficiency, making it an ideal model for modifier genetic screening to identify novel APOL1-RA interacting factors that could rescue APOL1 renal toxicity, or to test hundreds of drug compunds identified from cell-based APOL1 drug screens. We propose the following three aims to exploit these unique advantages of Drosophila for APOL1 nephropathy studies: Aim 1, Identify downstream genes of APOL1-RA using RNA-Seq and cross-species comparison, and test them as potential therapeutic targets using Drosophila nephrocytes and human podocytes; Aim 2, Identify modifier genes for APOL1 renal toxicity using Drosophila genetic screening, and test them as potential therapeutic targets in nephrocytes and human podocytes; Aim 3, Use Drosophila as a drug screening platform to screen positive hits from a cell-based APOL1 drug screen. Accomplishing the above proposed aims will yield novel, ground-breaking discoveries to understand molecular mechanisms of APOL1 nephropathy. We will identify down-stream APOL1-RA target genes, identify potential therapeutic targets using Drosophila genetic screening plus human podocyte validation and screen the most effective and least toxic drug compounds that may eventually be used to treat APOL1 nephropathy.

非裔美国人患肾脏疾病的风险较高，这在很大程度上是由于 APOL1 风险等位基因的遗传（RA）。 APOL1 肾病的分子和细胞机制正在得到广泛研究，但其机制仍不清楚，潜在的治疗靶点仍然隐藏。我们生成了果蝇研究 APOL1 肾病模型并表明 APOL1-RA 导致肾细胞（昆虫）的肾细胞损伤人类足细胞的结构和功能同源物。最近开发的小鼠 APOL1 模型提供是研究 APOL1 肾病分子机制和测试有希望的治疗方法的关键工具，但它不能用于进行大规模遗传筛选来识别可能拮抗肾功能的修饰基因 APOL1-RA 的毒性。用鼠标模型筛选数百个候选者也是极其困难的药物化合物。果蝇系统在低成本、高效率方面呈现出独特的优势，使其成为修饰遗传筛选的理想模型，以识别新的 APOL1-RA 相互作用因子，这些因子可以挽救 APOL1 肾毒性，或测试从基于细胞的 APOL1 药物中鉴定出的数百种药物化合物屏幕。我们提出以下三个目标来利用果蝇的这些独特优势用于 APOL1 肾病研究：目标 1，使用 RNA-Seq 和跨物种鉴定 APOL1-RA 下游基因比较，并使用果蝇肾细胞和人类足细胞测试它们作为潜在的治疗靶点；目标 2，使用果蝇遗传筛查鉴定 APOL1 肾毒性的修饰基因，并按如下方式进行测试：肾细胞和人类足细胞的潜在治疗靶点；目标3，利用果蝇作为药物筛选用于筛选基于细胞的 APOL1 药物筛选的阳性命中的平台。实现上述拟议目标将产生新颖的、突破性的发现，以了解 APOL1 肾病的分子机制。我们将鉴定下游 APOL1-RA 靶基因，利用果蝇遗传鉴定潜在的治疗靶点筛选加上人类足细胞验证并筛选最有效且毒性最小的药物化合物最终可能用于治疗 APOL1 肾病。