Discovery of small molecule promoters of cardiomyocyte proliferation to restore cardiac performance in disease

发现心肌细胞增殖的小分子启动子以恢复疾病中的心脏功能

• 批准号: 10397405
• 负责人: Alexandre Romain Colas
• 金额: $ 48.75万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-17 至 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10397405
• 关键词: Acute; Adult; Animal Model; Attention; Biological Assay; Biology; Blood Pressure; Bone Marrow; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Catalogs; Cell Lineage; Cells; Cellular Assay; Cellular biology; Chemicals; Chronic; Clinical; Critical Pathways; Data; Developed Countries; Developing Countries; Disease; Dose; Face; Fibroblasts; Foundations; Future; Generations; Goals; Grant; Heart; Heart Diseases; Heart Rate; Human; In Vitro; Investments; Lead; Lipids; Mammals; Mitotic; Myocardium; Natural regeneration; Neonatal; Neoplasms; Operative Surgical Procedures; Organ; Output; Parents; Pathway interactions; Patients; Performance; Persons; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Physiological; Pluripotent Stem Cells; Powder dose form; Prevention; Process; Proliferating; Property; Pump; Rattus; Readiness; Reporting; Research; Resources; Series; Structure-Activity Relationship; System; Technology; Testing; Therapeutic; Tissues; Transgenes; Validation; Work; analog; burden of illness; cardiac regeneration; cell type; cheminformatics; design; drug discovery; experimental study; heart function; high throughput screening; improved; in vivo evaluation; induced pluripotent stem cell; induced pluripotent stem cell derived cardiomyocytes; innovation; insight; liquid chromatography mass spectrometry; loss of function; metaplastic cell transformation; mortality; mouse model; neoplastic; novel; novel therapeutics; overexpression; promoter; prototype; response; restoration; risk minimization; scaffold; screening; small molecule; small molecule libraries; stem cells; tool; vascular risk factor

TITLE: Discovery of small molecule promoters of cardiomyocyte proliferation to restore cardiac performance in disease PROJECT SUMMARY Cardiovascular disease is the leading cause of mortality in developed and developing nations. Existing CVD pharmacotherapy options do not treat any foundational problems in the myocardium and its constituent cardiomyocytes; rather, unloading the heart and reducing vascular risk factors are the primary therapeutic strategies. Future strategies should move beyond prevention and treatment to restoration, regeneration, and replacement of functional cardiac tissue. Two fundamental challenges face the several mechanisms of potential cardiomyocyte renewal (proliferation of endogenous cardiomyocytes, resident progenitor cells, or extracardiac progenitor cells including bone-marrow derived or pluripotent cells): can a sufficient number of new cardiomyocytes be generated to meaningfully improve cardiac function in various diseases? And since most tissues in adult mammals remain post-mitotic to minimize the risk of cellular transformation into an uncontrolled proliferating neoplastic state, can cardiomyocyte-specific proliferation mechanisms be discovered and manipulated? It is unclear that the reported mechanisms of cardiomyocyte proliferation meet both of these challenges. We have recently developed a novel platform to conduct high throughput screening (HTS) on functional human cardiomyocytes matured from iPS cells. Our innovation is a hypothesis-free, phenotypic screening cascade designed to discover previously unknown, cardiomyocyte selective promoters of cardiomyocyte proliferation. Our hypothesis is this approach will ultimately generation drug-like starting points for future disease-modifying cardiovascular therapeutics. The primary HTS assay has been fully optimized in a 384-well format, and as a demonstration of assay readiness, 12,000 compounds have been screened (Z’- factor of 0.4). Multiple hits from pilot screens were identified and were confirmed and validated in concentration response experiments. A battery of downstream assays has been developed to establish a critical path-testing funnel. Several compounds identified from the pilot screen specifically promoted cardiomyocyte proliferation versus fibroblast proliferation, and furthermore promoted ex vivo proliferation in both neonatal and adult cardiomyocytes isolated from rats. This proposal builds on data from the applicants, an established team from SBP (Drs. Larson and Colas) with basic biology and drug discovery expertise in the field and access to all necessary technologies. The overall goal of this proposal is to generate chemical biology research tools and the starting points for new drugs. As the critical path assays are all in place, we anticipate we can rapidly obtain such probe molecules and start to explore their activity. Our future plans beyond this grant are to ultimately determine their suitability for hit-to-lead activities, begin in vivo evaluation of lead compounds in animal models and eventually patients, and determine their cellular mechanism of action. The product of the work proposed in this application will serve as preliminary data for both hit-to-lead (HTL) grant submissions and parent R01 grant submissions to pursue basic biology understanding of their mechanisms.

标题：发现心肌细胞增殖的小分子促进剂以恢复心脏功能 疾病表现 项目摘要 心血管疾病是发达国家和发展中国家死亡的主要原因。现有CVD 药物治疗方案不能治疗心肌及其组成部分的任何基础问题， 心肌细胞;相反，卸载心脏和减少血管危险因素是主要的治疗方法。 战略布局未来的战略应该从预防和治疗转向恢复、再生和重建。 替换功能性心脏组织。两个基本的挑战面临的几个机制， 潜在的心肌细胞更新（内源性心肌细胞、驻留祖细胞或 心外祖细胞，包括骨髓来源的或多能细胞）：足够数量的 产生新的心肌细胞来有意义地改善各种疾病的心脏功能？既然 成年哺乳动物的大部分组织保持在有丝分裂后， 不受控制的增殖肿瘤状态，是否可以发现心肌细胞特异性增殖机制 被人操纵目前尚不清楚所报道的心肌细胞增殖的机制是否满足这两个条件 挑战我们最近开发了一种新的平台，用于对 从iPS细胞成熟的功能性人心肌细胞。我们的创新是一种无假设的， 筛选级联设计发现以前未知的，心肌细胞选择性启动子， 心肌细胞增殖我们的假设是这种方法最终会产生类似药物的起点 用于未来改善疾病的心血管疗法。主要HTS检测试剂盒已在 384-孔格式，并且作为测定准备就绪的证明，已经筛选了12，000种化合物（Z '- 系数0.4）。鉴定了中试筛选的多个命中，并在浓度中进行了确认和验证 反应实验。已经开发了一系列下游检测方法来建立关键路径测试 漏斗从初步筛选中鉴定出的几种化合物特异性地促进心肌细胞增殖 与成纤维细胞增殖，并进一步促进新生儿和成人的离体增殖 从大鼠分离的心肌细胞。该提案建立在申请人的数据基础上， SBP（Drs. Larson和Colas）具有该领域的基础生物学和药物发现专业知识，并可访问所有 必要的技术。该提案的总体目标是生成化学生物学研究工具， 新药的起点。由于关键路径分析都已到位，我们预计我们可以迅速 获得这样的探针分子并开始探索它们的活性。除了这笔赠款，我们的未来计划是 最终确定其对铅活性的适用性，开始体内评价铅化合物， 动物模型，最终是患者，并确定它们的细胞作用机制。的产物 本申请中提出的工作将作为两个点击到铅（HTL）赠款提交的初步数据 和父R 01资助提交，以追求其机制的基本生物学理解。