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

非裔美国人面临肾脏疾病的风险增加，这在很大程度上是由于APOL 1风险等位基因的遗传 （RA）。APOL 1肾病的分子和细胞机制正在被广泛研究， 但其机制尚不清楚，潜在的治疗靶点仍不清楚。我们创造了一个果蝇 研究APOL 1肾病模型，并显示APOL 1-RA导致肾细胞损伤，昆虫 人足细胞的结构和功能同源物。最近开发的小鼠APOL 1模型提供了 是研究APOL 1肾病分子机制和测试有前景的治疗方法的关键工具，但它 不能用于进行大规模的遗传筛选，以确定修饰基因，可以拮抗肾 APOL 1-RA的毒性。使用鼠标模型筛选数百名候选人也是极其困难的 药物化合物。果蝇系统在低成本、高效率方面具有独特的优势， 使其成为修饰基因筛选的理想模型，以鉴定新的APOL 1-RA相互作用因子， 拯救APOL 1肾毒性，或测试从基于细胞的APOL 1药物中鉴定的数百种药物化合物 卡位我们提出了以下三个目标，以利用果蝇对APOL 1的这些独特优势 肾病研究：目的1，使用RNA-Seq和跨物种鉴定APOL 1-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; 目的2：利用果蝇基因筛选技术，鉴定APOL 1肾毒性修饰基因，并进行验证。 肾细胞和人类足细胞中的潜在治疗靶点;目的3，利用果蝇作为药物筛选 该平台用于从基于细胞的APOL 1药物筛选中筛选阳性命中。实现上述目标 将产生新的，突破性的发现，以了解APOL 1肾病的分子机制。我们 将确定下游APOL 1-RA靶基因，使用果蝇遗传学方法确定潜在的治疗靶点， 筛选加上人类足细胞验证，并筛选最有效和毒性最小的药物化合物， 可能最终用于治疗APOL 1肾病。

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.