Discovering ADPKD Modifier Genes and Therapies via Zebrafish Genetics

通过斑马鱼遗传学发现 ADPKD 修饰基因和疗法

• 批准号: 10915786
• 负责人: XUEYING LIN
• 金额: $ 10万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2024-09-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10915786
• 关键词: Adult; Affect; Animal Model; Animals; Autosomal Dominant Polycystic Kidney; Biological Assay; Cyst; Cystic kidney; Data; Development; Disease Progression; Disease model; Embryo; Exhibits; Fishes; Foundations; Future; Genes; Genetic; Genetic Diseases; Genetic Identity; Genetic Screening; Genetic screening method; Homologous Gene; Human; Kidney; Lead; Life; Measurement; Measures; Mediating; Methods; Modeling; Molecular; Parents; Patients; Phase; Phenotype; Play; Population; Protocols documentation; Role; Sampling; Signal Pathway; Solid; Study models; Technology; Testing; Zebrafish; gene discovery; human disease; in vivo; mutant; novel; patient population; rapid technique; screening; tool; zebrafish genome

ADPKD patients exhibit highly variable phenotypes, of which genetic modifiers were thought to play an important role. However, the identity of the genetic modifiers remains largely unknown, because of the lack of an efficient method. In this context, here, we propose to establish adult zebrafish as a new animal model that enables rapid and systematic discovery of genetic modifiers of ADPKD. We have generated substantial preliminary data for this proposal: 1) proving the molecular conservation of the zebrafish model by identifying corresponding homologues for 99% of the 82 PKD-associated genes; 2) developing zebrafish mutants for pkd1, pkd2 and ift140, 3 of the 6 known ADPKD-associated genes, which all lead to renal cyst formation; 3) demonstrating the feasibility of utilizing a novel microhomology-mediated end joining (MMEJ)-based F0 genetic method for screening ADPKD modifier genes. These preliminary data prompted us to raise a central hypothesis predicting that zebrafish is a conserved and efficient in vivo vertebrate model for discovering genetic modifiers of ADPKD. To test this hypothesis, we proposed the following three specific aims. In Aim 1, we will further prove the feasibility of our strategy by validating 2 novel protective ADPKD modifiers emerged from a MMEJ-based pilot F0 genetic screen. In Aim 2, we will explore the feasibility of using zebrafish models to assess ADPKD modifiers suggested from human patient population. In Aim 3, we will facilitate the use of zebrafish for PKD studies by further developing NMR protocols for the measurement of zebrafish kidney volume and cysts. Upon completion of the proposal, we anticipate to significantly strengthen the parent proposal, aiming to establish a F0-based genetic screen approach for systematically identifying new modifier genes for ADPKD. The launch of this novel animal model is expected to significantly advance the ADPKD field.

ADPKD患者表现出高度可变的表型，其中遗传修饰剂被认为起着重要作用。然而，由于缺乏有效的方法，遗传修饰剂的身份在很大程度上仍然未知。在这种情况下，在这里，我们建议建立成年斑马鱼作为一种新的动物模型，使快速和系统的发现ADPKD的遗传修饰剂。我们已经为这一提议提供了大量的初步数据： 1)通过鉴定82个PKD相关基因中99%的相应同源物，证明了斑马鱼模型的分子保守性; 2）开发了6个已知ADPKD相关基因中的3个pkd 1、pkd 2和ift 140的斑马鱼突变体，这些基因都导致肾囊肿形成; 3）证明了利用基于微同源介导的末端连接（MMEJ）的F0遗传学方法筛选ADPKD修饰基因的可行性。这些初步的数据促使我们提出一个中心假设，预测斑马鱼是一个保守的和有效的体内脊椎动物模型，发现ADPKD的遗传修饰。为了验证这一假设，我们提出了以下三个具体目标。在目标1中，我们将通过验证从基于MMEJ的试点F0遗传筛选中出现的2种新型保护性ADPKD修饰剂来进一步证明我们策略的可行性。在目标2中，我们将探索使用斑马鱼模型来评估从人类患者群体中提出的ADPKD调节剂的可行性。在目标3中，我们将通过进一步开发用于测量斑马鱼肾脏体积和囊肿的NMR协议来促进斑马鱼用于PKD研究。该提案完成后，我们预计将大大加强母提案，旨在建立一种基于F0的遗传筛选方法，用于系统地识别ADPKD的新修饰基因。这种新型动物模型的推出预计将显着推进ADPKD领域。