Discovering cardiomyopathy modifiers and therapies via zebrafish genetics

项目摘要 心肌病是一种具有高度异质性的疾病：>100个基因与不同类型相关 心肌病，如肥厚型心肌病（HCM）和扩张型心肌病（DCM）;患者 具有相同突变的人可能会表现出高度不同的疾病发作和严重程度，大概是因为 不同的遗传和环境因素。然而，遗传修饰剂的身份在很大程度上仍然是 未知为了满足这一需求，我们的团队一直在领导成年斑马鱼作为脊椎动物的发展。 心肌病模型。在前两个融资周期中，我们开发了一种新的前向遗传学， 筛选策略，以发现阿霉素（DOX）诱导的心肌病（DIC）的修饰基因。在这里， 我们计划将这种方法扩展到遗传性心肌病。我们已经产生了大量的初步 数据来证明该建议的可行性，包括鉴定5个原始DIC修饰基因和4个 另外的候选DIC修饰基因，BAG 3心肌病的斑马鱼模型的产生， mTOR作为治疗性修饰基因的鉴定，以及胚胎鱼-成鱼- 小鼠药物评估平台。总之，这些数据促使我们测试这个中心假设， 该提案预测，遗传性心肌病模型的修饰基因可以通过 斑马鱼的正向遗传策略，从中可以获得治疗靶基因和相关化合物， 被高效的斑马鱼遗传学迅速发现。该提案分为两个具体目标。在特定 目的1，我们将测试假设，基于遗传学的方法是可扩展到bag 3 以确定治疗性修饰剂。我们将确定表型进展和变异， 斑马鱼bag 3 KO，评估9种DIC调节剂对bag 3心肌病的调节作用，然后鉴定 Bag 3心肌病的治疗修饰剂。我们将通过转录组来阐明潜在的机制 分析.在具体目标2中，我们将阐明mTOR治疗作用的潜在机制。 抑制，证实了bag 3心肌病基于自噬疗法，并重新利用FDA批准的自噬- 激活药物治疗Bag 3心肌病。据估计，这一新战略的发展， 该提案将显著推进预后测试开发、风险分层和个性化治疗。 心肌病

Project Summary Cardiomyopathy is a disorder with high heterogeneity: >100 genes have been linked to different types of cardiomyopathy, such as hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM); patients with the same mutation can manifest highly variable disease onset and severity, presumably because of different genetic and environmental factors. However, the identity of genetic modifiers remains largely unknown. To address the need, our team has been leading the development of adult zebrafish as a vertebrate model for cardiomyopathy. During the previous 2 funding cycles, we developed a novel forward genetic screening strategy for discovering modifier genes for doxorubicin (DOX)–induced cardiomyopathy (DIC). Here, we plan to extend this approach to inherited cardiomyopathies. We have generated substantial preliminary data to prove the feasibility of the proposal, including the identification of 5 original DIC modifier genes and 4 additional candidate DIC modifier genes, the generation of a zebrafish model of BAG3 cardiomyopathy, the identification of mtor as a therapeutic modifier gene, and the establishment of an embryonic fish-adult fish- mouse drug assessment platform. Together, these data prompted us to test the central hypothesis of this proposal, which predicts that modifier genes for an inherited cardiomyopathy model can be identified via a forward genetic strategy in zebrafish, from which therapeutic target genes and related compounds can be rapidly discovered by efficient zebrafish genetics. The proposal is organized into 2 specific aims. In Specific Aim 1, we will test the hypothesis that a forward genetics-based approach is extendable to bag3 cardiomyopathy to identify therapeutic modifiers. We will determine phenotypic progression and variation in zebrafish bag3 KO, assess modifying effects of 9 DIC modifiers on bag3 cardiomyopathies, and then identify therapeutic modifiers for bag3 cardiomyopathy. We will elucidate underlying mechanism by transcriptome analysis. In Specific Aim 2, we will elucidate underlying mechanisms of the therapeutic effects of mTOR inhibition, prove autophagy-based therapy for bag3 cardiomyopathy, and repurpose FDA-approved autophagy- activating drugs to treat bag3 cardiomyopathy. It is anticipated that the novel strategy developed by this proposal will significantly advance prognostic test development, risk stratification, and personalized therapy for cardiomyopathies.