Modeling Nephrotic Syndrome in Drosophila Nephrocytes

果蝇肾细胞肾病综合征建模

基本信息

批准号：
10019519
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：
2014
资助国家：
美国
起止时间：
2014-04-10 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10019519
关键词：
Actins Affect Animal Model Autophagocytosis Autophagosome Biological Biological Models Biology Cell physiology Child Collaborations Communities Complement Cytoskeleton DNA Sequence Alteration DNA sequencing Data Defect Disease Disease model Drosophila genus Drug Screening Drug Targeting End stage renal failure FRAP1 gene Funding Gene Mutation Gene Silencing Genes Genetic Genomics Grant Human Kidney Kidney Diseases Label Methods Modeling Molecular Structure Mus Mutation Nephrotic Syndrome Pathogenesis Pathogenicity Pathway interactions Patients Pharmaceutical Preparations Phenotype Play Process Regulation Renal function Renal glomerular disease Reproducibility Research Role Series Signal Pathway Steroid-resistant idiopathic nephrotic syndrome Study models System Technology Testing Therapeutic Effect Time Transgenic Organisms Validation Variant Work Zebrafish base cohort cost disease mechanisms study drug testing effective therapy fly gene replacement genetic variant in vivo inhibitor/antagonist innovation kidney cell live cell imaging novel novel therapeutics podocyte rare genetic disorder screening small molecule targeted treatment tool young adult

项目摘要

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.

项目概要/摘要：肾病综合征 (NS) 是儿童终末期肾病 (ESRD) 的最常见原因之一和年轻人，但缺乏有效的治疗方法，特别是对于类固醇抵抗性肾病综合征（SRNS）。 DNA测序技术的快速发展已经导致大量的DNA鉴定遗传变异是 SRNS 的潜在致病因素。然而，缺乏这些的体内功能数据候选 SRNS 基因及其变异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精确突变的动物模型，用于疾病机制研究和测试潜在的靶向治疗的需求很大。我们建立了低成本、高效率的果蝇模型系统生成候选 NS 基因和变异的基本功能数据，并加快新 NS 基因的鉴定。这种新颖的肾脏疾病模型系统利用了卓越的分子、果蝇肾细胞和人类足细胞的结构和功能等同。我们研究了 40 个已知的肾细胞中的 NS 基因，发现这些基因中有 85% 在从果蝇到肾细胞的肾细胞中发挥保守作用人类。我们还通过生成个性化果蝇 NS 模型发现了潜在的疾病机制哪些内源性果蝇基因在功能上被携带患者来源的人类同源物取代突变。我们还利用这些fly NS模型开发了药物测试平台，并成功逆转了使用根据疾病机制告知的靶向治疗来确定肾脏表型。在此更新提案中，我们将使用果蝇中用于识别参与自噬和细胞骨架的新肾脏基因的强大遗传工具规定。我们将鉴定新的肾细胞细胞骨架标记物和成分。我们还将开发新的候选 NS 基因的个性化果蝇模型和已知 NS 基因的新遗传变异，如以及使用飞行模型来测试潜在的靶向疗法。我们的研究将提供肾脏疾病研究界拥有低成本、高效的模型系统，可对 NS 相关基因进行功能验证和遗传变异，以确定新的 NS 基因，并开发基于机制的靶向治疗。