Modeling Nephrotic Syndrome in Drosophila Nephrocytes

项目摘要/摘要： 肾病综合征(NS)是儿童终末期肾病(ESRD)最常见的病因之一 和年轻人，但缺乏有效的治疗，特别是对激素抵抗型肾病综合征 (SRNS)。DNA测序技术的快速进步导致了大量 是SRNS潜在致病因素的遗传变异。然而，缺乏这些细胞在体内的功能数据 候选SRNS基因及其变体使得验证它们在导致疾病中的作用变得困难。一个 携带在患者身上发现的准确突变的动物模型，用于疾病机制研究和测试 潜在的靶向治疗需求很大。我们已经建立了一种低成本、高效率的果蝇 模型系统，以生成候选NS基因和变体的基本功能数据，并加快 新NS基因的鉴定。这一新的肾脏疾病模型系统利用了非凡的分子， 果蝇肾细胞和人足细胞的结构和功能等价性。我们研究了40个已知的 NS基因，发现其中85%的基因在苍蝇肾细胞中起保守作用。 人类。我们还通过生成个性化的Fly NS模型发现了潜在的疾病机制 哪些内源性苍蝇基因在功能上被携带患者来源的人类同源基因取代 突变。我们还利用这些Fly NS模型开发了药物测试平台，并成功地逆转了 根据发病机制进行靶向治疗的肾脏表型。在此续订建议书中，我们将使用 在果蝇中识别与自噬和细胞骨架有关的新的肾脏基因的强大遗传工具 监管。我们将确定新的肾细胞细胞骨架标志物和成分。我们还将开发新的 候选NS基因和已知NS基因的新遗传变体的个性化果蝇模型，如 以及使用苍蝇模型来测试潜在的靶向治疗。我们的研究将提供肾脏疾病 使用低成本、高效的模型系统从功能上验证NS相关基因的研究社区 和遗传变异，以确定新的NS基因，并开发基于机制的靶向治疗。

PROJECT SUMMARY/ABSTRACT: Nephrotic syndrome (NS) is one of the most frequent causes of End-Stage Renal Disease (ESRD) in children and young adults, but effective treatment is lacking, particularly for Steroid-Resistant Nephrotic Syndrome (SRNS). Rapid advances in DNA sequencing technology have led to the identification of large numbers of genetic variants that are potential causal factors for SRNS. However, lack of in vivo functional data for these candidate SRNS genes and their variants make it difficult to validate their roles in causing the disease. An animal model that carry the exact mutation found in patients for disease mechanism studies and testing of potential targeted therapies is in great demand. We have established a low-cost, high-efficiency Drosophila model system to generate essential functional data for candidate NS genes and variants, and to expedite the identification of novel NS genes. This novel kidney disease model system exploits the remarkable molecular, structural and functional equivalencies of Drosophila nephrocytes and human podocytes. We studied 40 known NS genes in nephrocytes and found that 85% of these genes play conserved roles in kidney cells from flies to humans. We also discovered underlying disease mechanisms by generating personalized fly NS models in which endogenous fly genes were functionally replaced by human homologs carrying patient-derived mutations. We also developed drug testing platform using these fly NS models, and successfully reversed the renal phenotype using targeted therapy informed by disease mechanism. In this renew proposal, we will use the powerful genetic tools in Drosophila to identify new renal genes involved in autophagy and cytoskeleton regulation. We will identify new nephrocyte cytoskeleton markers and components. We will also develop new personalized Drosophila models for candidate NS genes and novel genetic variants for known NS genes, as well as using the fly models to test potential targeted therapies. Our studies will provide the kidney disease research community with a low-cost high-efficiency model system to functionally validate NS associated genes and genetic variants, to identify novel NS genes, and to develop mechanism-based targeted therapies.