Project 2 - Shh and Etv2 Signaling Pathways and Cardiovascular Repair in Mouse and Pig

项目 2 - Shh 和 Etv2 信号通路以及小鼠和猪的心血管修复

• 批准号: 10677734
• 负责人: Daniel J. Garry
• 金额: $ 43万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677734
• 关键词: Adult; Algorithms; Animal Model; Applications Grants; Bioinformatics; Birth; Blood Vessels; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Cell Proliferation; Clinical Research; Collaborations; Complement; Data; Development; Dissection; Endothelium; Engineering; Family suidae; Future; Gene Expression; Genetic; Genetic Engineering; Genetic Models; Goals; Growth; Heart; Heart Diseases; Heart Injuries; Heart failure; In Vitro; Injury; Ischemia; Knock-out; Laboratories; Lower Organism; Messenger RNA; Molecular; Molecular Analysis; Morbidity - disease rate; Mus; Muscle; Myocardial Infarction; Natural regeneration; Neonatal; Newts; Pathway interactions; Patient Care; Patients; Proliferating; Publications; Publishing; Regenerative capacity; Role; SOX4 gene; Signal Pathway; Societies; Techniques; Technology; Testing; Therapeutic; Tissues; Ventricular; Viral Vector; angiogenesis; cardiac regeneration; cardiac repair; clinically significant; congenital heart disorder; experimental study; heart cell; injured; innovation; mortality; mouse model; myocardial injury; novel; novel therapeutics; overexpression; porcine model; postnatal; pre-clinical; programs; repaired; synergism; tool

ABSTRACT Cardiovascular disease is common and deadly. While lower organisms have a tremendous capacity for regeneration, the adult mammalian heart is more limited in its capacity for regeneration and remuscularization of the damaged tissue following an injury such as a myocardial infarction. Recently, we defined an evolutionary conserved Shh signaling pathway in newt and mouse that promotes cardiomyocyte proliferation in vitro and cardiac repair. Our preliminary data support the hypothesis that Gli1 (a downstream effector of Shh) is a direct upstream regulator of Sox4 gene expression and Gli1 overexpression promotes cardiomyocyte proliferation in vitro. Furthermore, we have demonstrated that Etv2 is a master regulator of the endothelial lineage and is necessary and sufficient for angiogenesis in mouse and pig. Importantly, the results of these studies provide a platform for the proposed studies that will dissect the mechanisms that govern cardiovascular repair and proliferation. Therefore, our overall hypothesis is that the Shh downstream effectors, Gli1/Gli2, regulate cardiomyocyte proliferation in a Sox4 dependent manner and Etv2 promotes endothelial/vascular development to collectively promote cardiac regeneration. In these proposed studies, we will use a number of novel genetic models and large animal models that are available in our laboratory (and the cores), bioinformatics algorithms that we developed and we take an innovative approach to dissect the role of Gli1, Etv2 and Sox4 as important factors that govern cardiovascular repair and proliferation. To examine our hypotheses, the revised Project 2 proposal will address the following specific aims: Specific Aim #1: To define the capacity of Gli and Etv2 deficient hearts for growth and regeneration; Specific Aim #2: To determine the impact of Gli1 and Etv2 overexpression on cardiac repair and regeneration and Specific Aim #3: To define the mechanistic role of Sox4 in the cardiomyocyte lineage. These aims will complement and synergize with the other projects. They will utilize our recently engineered genetic mouse models, inducible viral vectors, modified mRNA-GFP constructs, mouse and pig models, molecular analyses and bioinformatics algorithms to comprehensively define the role for Gli1, Etv2 and Sox4 as essential factors that govern cardiac repair and regeneration by promoting cardiomyocyte proliferation and angiogenesis following injury. Given the tremendous morbidity and mortality of cardiovascular disease in our society, the potential impact of this proposal is tremendous.

抽象的 心血管疾病是常见且致命的。虽然低等生物具有巨大的能力 再生，成年哺乳动物心脏的再生和重新肌肉化的能力更加有限 心肌梗塞等损伤后受损组织的修复。最近，我们定义了一个进化 蝾螈和小鼠中保守的Shh信号通路在体外促进心肌细胞增殖 心脏修复。我们的初步数据支持这样的假设：Gli1（Shh 的下游效应器）是直接 Sox4基因表达和Gli1过表达的上游调节因子促进心肌细胞增殖 体外。此外，我们已经证明 Etv2 是内皮细胞谱系的主要调节因子，并且 对于小鼠和猪的血管生成是必要且充分的。重要的是，这些研究的结果提供了一个 拟议研究的平台将剖析控制心血管修复和 增殖。因此，我们的总体假设是 Shh 下游效应器 Gli1/Gli2 调节 心肌细胞以 Sox4 依赖性方式增殖，Etv2 促进内皮/血管 共同促进心脏再生的发展。在这些拟议的研究中，我们将使用 我们实验室（和核心）提供的新型遗传模型和大型动物模型的数量， 我们开发的生物信息学算法，我们采用创新方法来剖析 Gli1 的作用， Etv2 和 Sox4 是控制心血管修复和增殖的重要因子。来检查我们的 根据假设，修订后的项目 2 提案将解决以下具体目标： 具体目标 1： 确定 Gli 和 Etv2 缺陷心脏的生长和再生能力；具体目标#2： 确定 Gli1 和 Etv2 过表达对心脏修复和再生的影响以及特异性 目标#3：确定 Sox4 在心肌细胞谱系中的机制作用。这些目标将 与其他项目相互补充和协同。他们将利用我们最近改造的基因小鼠 模型、诱导型病毒载体、修饰的 mRNA-GFP 构建体、小鼠和猪模型、分子分析 和生物信息学算法全面定义 Gli1、Etv2 和 Sox4 作为重要因素的作用 通过促进心肌细胞增殖和血管生成来控制心脏修复和再生 受伤后。鉴于心血管疾病在我们社会中的巨大发病率和死亡率， 该提案的潜在影响是巨大的。