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: 10611642
• 负责人: Alexandre Romain Colas
• 金额: $ 0.4万
• 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/10611642
• 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启动子以恢复心脏 疾病表现 项目摘要 心血管疾病是发达国家死亡率的主要原因。现有的CVD 药物治疗方案没有治疗心肌及其成分中的任何基本问题 心肌细胞；相反，卸载心脏并减少血管危险因素是主要疗法 策略。未来的策略应超越预防和治疗，以恢复，再生和 替代功能性心脏组织。两个基本挑战面临着几种机制 潜在的心肌细胞更新（内源性心肌细胞的增殖，常驻祖细胞或 心脏外祖细胞，包括衍生或多能细胞，包括骨髓）：足够数量的数量 生成新的心肌细胞以有意义地改善各种疾病中的心脏功能？从那以后 成年哺乳动物中的大多数组织仍然在有丝分裂后，以最大程度地减少细胞转化为 不受控制的增生肿瘤状态，可以发现心肌细胞特异性增殖机制 并操纵？目前尚不清楚所报道的心肌细胞增殖机制符合这两个 挑战。我们最近开发了一个新颖的平台，以对高吞吐量筛查（HTS）进行 从IPS细胞成熟的功能性人类心肌细胞。我们的创新是一种无假设的表型 筛选级联 心肌细胞增殖。我们的假设是这种方法最终将产生类似毒品的起点 用于改善疾病的心血管疗法。主要的HTS测定已在A中进行了完全优化 384孔格式，作为组装准备的演示，已经筛选了12,000种化合物（Z'- 因子为0.4）。鉴定了来自飞行员屏幕的多次命中，并以浓度进行了确认并验证 响应实验。已经开发了一系列下游测定，以建立关键的路径测试 漏斗。从试验筛查中鉴定出的几种化合物专门促进了心肌细胞增殖 与成纤维细胞增殖相比，此外，新生儿和成人促进了离体增殖 从大鼠分离的心肌细胞。该提案以申请人的数据为基础，这是一个已建立的团队 具有基本生物学和药物发现专业知识的SBP（Larson和Colas博士） 必要的技术。该建议的总体目标是生成化学生物学研究工具和 新药的起点。由于关键的路径断言都到位，我们预计我们可以迅速 获得此类探针分子并开始探索其活性。我们的未来计划以外的赠款是 最终确定其对命中率活动的适用性，开始对铅化合物的体内评估 动物模型，有时是患者，并确定其细胞作用机理。乘积 本申请中提出的工作将作为命中率高（HTL）赠款提交的初步数据 父母R01赠款提交，以购买对其机制的基本生物学理解。

项目成果

Genetic architecture of natural variation of cardiac performance from flies to humans.

• DOI: 10.7554/elife.82459
• 发表时间: 2022-11-16
• 期刊: eLife
• 影响因子: 7.7
• 作者: Saha S;Spinelli L;Castro Mondragon JA;Kervadec A;Lynott M;Kremmer L;Roder L;Krifa S;Torres M;Brun C;Vogler G;Bodmer R;Colas AR;Ocorr K;Perrin L
• 通讯作者: Perrin L