Pediatric Heart Disease: Getting from Mutations to Therapeutics

小儿心脏病：从突变到治疗

• 批准号: 10112285
• 负责人: BRUCE D GELB
• 金额: $ 86.08万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-15 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10112285
• 关键词: ADME Study; Address; Adopted; Adult; Animals; Back; Biological; Biological Models; Cardiac; Cardiovascular Diseases; Caring; Cell Line; Cells; Chemicals; Childhood; Clinical Trials Design; Complex; Congenital Heart Defects; Data; Development; Disease; Disease model; Drosophila genus; Drosophila melanogaster; Drug Interactions; Drug usage; Eye; FDA approved; Genetic; Genetic Diseases; Genomics; Heart Diseases; Heart Hypertrophy; Homeostasis; Human; Hypertrophic Cardiomyopathy; Libraries; Methods; Mission; Mitogen-Activated Protein Kinases; Modeling; Mutant Strains Mice; Mutation; Myocardial; National Heart, Lung, and Blood Institute; Noonan Syndrome; Other Genetics; Pathogenesis; Pharmaceutical Preparations; Pharmacogenomics; Pharmacology; Phenotype; Plant Roots; Premature Mortality; Pupa; RAF1 gene; Research Personnel; Robotics; Signal Transduction; System; Technology; Testing; Therapeutic; Toxic effect; Veins; Vial device; Wing; accurate diagnosis; base; causal variant; comorbidity; disease-causing mutation; drug development; fly; gene discovery; high throughput screening; improved; induced pluripotent stem cell; interest; mouse model; mutant; novel therapeutics; prognostic; screening; side effect; small molecule; small molecule libraries; stem cell model; trait

Pediatric cardiovascular disorders, which comprise congenital heart defects (CHD) and myocardial and conduction system diseases, remain highly challenging due to cardiac co-morbidities and premature mortality. As most of these disorders are genetic, efforts over the past 30 years have focused on identifying their causal mutations. Particularly for Mendelian traits such as Noonan syndrome and related disorders (the RASopathies), this has been highly successful. Newer genomic technologies have accelerated gene discovery for pediatric cardiovascular disorders, including genetically complex ones. These genetic discoveries are improving care through more accurate diagnosis, better prognostication, and refinement of clinical trial design. What has not occurred with rare exception is the development of novel therapies based on the new understanding of disease pathogenesis enabled by these gene discoveries. Finding therapies for pediatric cardiovascular disorders will be challenging because the biological targets are generally central to cell homeostasis (e.g., RAS/MAP kinase signaling) so cannot be completely inhibited for long periods without incurring side effects that would outweigh their benefits. For this R35 mechanism, I and my outstanding co- investigators with relevant expertise intend to address this gap using a drug development pipeline that begins with high-throughput screening to overcome pupal lethality in Drosophila melanogaster models of disease with a chemical library that covers druggable space (n=14,400) using 96-well plates and robotics. Screening in whole animals is performed agnostically and has the putative advantage of providing a simultaneous read out of efficacy and toxicity. We provide preliminary data showing that we have already achieved this using a fly RAF1 mutant model of Noonan syndrome with hypertrophic cardiomyopathy. Subsequent steps with fruit flies include confirmation of initial hits in vials, determining efficacy against adult fly phenotypes such as rough eye, ectopic wing veins and heart hypertrophy. Back-up libraries for the candidate compounds, typically 60-80 chemical neighbors, will be culled for ones with most desirable drug traits and then screened in the fly models. Using a defined set of fruit fly deficiency lines, targets and anti-targets will be established to enable further rounds of rational pharmacology. ADME studies will be used to reduce potential for drug-drug interactions. In parallel, we will pursue repurposing of FDA-approved drugs using library screening with fruit fly models and systems pharmacogenomics. Leading compounds and drugs will then be tested against phenotypes in human induced pluripotent stem cell lines with the disease-causing mutation for efficacy. The most promising drugs will then be tested in existing mouse models (e.g., HCM in Raf1 mutant mice) using appropriate endpoints. Taken as a whole, the approach proposed will significantly advance the identification of novel therapeutics for pediatric cardiovascular diseases, starting with the RASopathies and later for other traits. If robust, this will provide a paradigm that can be adopted for other genetic traits of interest to the NHLBI.

小儿心血管疾病，包括先天性心脏缺陷（CHD）和心肌和心肌和 传导系统疾病，由于心脏合并症和过早死亡率，仍然高度挑战。 由于这些疾病大多数都是遗传的，因此过去30年中的努力集中在确定其因果关系上 突变。特别是对于孟德尔综合症和相关疾病等孟德尔的特征（ Rasopathies），这非常成功。较新的基因组技术加速了基因发现 用于儿科心血管疾病，包括遗传复杂的疾病。这些遗传发现是 通过更准确的诊断，更好的预后和临床试验设计的完善来改善护理。 在罕见的例外没有发生的是基于新的新疗法的开发 了解这些基因发现能够理解疾病发病机理。寻找疗法 小儿心血管疾病将具有挑战性，因为生物学靶标通常是细胞的核心 体内平衡（例如RAS/MAP激酶信号传导）因此长期无法完全抑制 会产生副作用，这将超过其利益。对于这种R35机制，我和我的出色共同 具有相关专业知识的调查人员打算使用开始的药物开发管道来解决这一差距 通过高通量筛查，以克服果蝇疾病模型中的pupal杀伤力 使用96孔板和机器人技术覆盖可毒空间（n = 14,400）的化学文库。筛选 整体动物的表现不足，并且具有同时读取的推定优势 功效和毒性。我们提供初步数据，表明我们已经使用苍蝇实现了这一点 Noonan综合征与肥厚性心肌病的RAF1突变模型。随后的果蝇步骤 包括确认小瓶中的初始命中，确定针对成年蝇型（例如粗眼）的功效 异位机翼静脉和心脏肥大。候选化合物的备用库，通常为60-80 化学邻居将为具有最理想的药物性状的邻居淘汰，然后在飞行模型中进行筛选。 使用一组定义的果蝇缺乏线，将建立目标和反靶标，以进一步 理性药理学回合。 ADME研究将用于减少药物相互作用的潜力。在 平行于，我们将使用果蝇模型和 系统药物基因组学。然后，将针对人类的表型测试铅化合物和药物 引起多能干细胞系具有致病性突变的功效。最有希望的药物 然后，将使用适当的端点在现有鼠标模型（例如RAF1突变小鼠中的HCM）中进行测试。 总体而言，提出的方法将显着促进对新型治疗剂的识别 小儿心血管疾病，从肉饼开始，然后是其他特征。如果强大的话，这将 提供一个可以用于NHLBI感兴趣的其他遗传特征的范式。