Cardiovascular regeneration and pioneer factors

心血管疾病既常见又致命。例如，外周动脉疾病影响更大 1000多万美国人每年导致超过15万人截肢。此外， 30多万名患者接受了冠状动脉搭桥术(外科血管重建术)。现代医学 血管疾病的治疗方法包括截肢和血管旁路移植--这些都是治疗性的 干预措施有很大的局限性。这些疾病是慢性的、使人衰弱的、致命的，它们需要新的 和新奇的疗法。以前的研究已经证明了先锋因素的基本作用 染色质的可及性，从而影响早期转录调节因子对谱系的结合 规格。我们最近证明ETV2是血管内皮细胞、血管内皮细胞的重要先驱因子。 和血统。我们使用了全局的和有条件的基因破坏策略，命运图，基因 编辑、单细胞RNA-SEQ、ATAC-SEQ和CHIP-SEQ分析，为这一应用提供支持数据。在……里面 此外，我们还定义了一个重要的ETV2-miR130a-PDGFRA级联，它调控内皮细胞的发育。 此外，我们最近的出版物和我们的初步数据支持总体假设 ETV2是调节内皮细胞系规范的先驱因子。在这些建议中 研究中，我们将使用我们设计的一些独特的遗传模型，我们将采取创新的 确定ETV2作为先导因子调节心血管功能的机制的策略 再生。为了检验我们的假设，我们将讨论以下具体目标：具体目标1： 具体目标#1：进一步确定ETV2作为先锋因素在 胚胎发生和内皮细胞谱系的重编程；具体目标2：确定 染色质修饰因子和ETV2在胚胎发育和内皮重编程中的作用 血统和特定目标#3：检查促进ETV2介导的重编程的因素 体外和体内的内皮细胞谱系。这些目标将利用我们最近改造的遗传小鼠 模型，ATAC-seq，MNase-seq，ChIP-seq，诱导性小鼠模型，成年小鼠心脏损伤模型， 新的生物信息学算法，全面定义ETV2作为 先锋因素，并将作为治疗倡议的前奏，以设计和促进再生 心血管血统。鉴于我国心血管疾病的巨大发病率和死亡率 对于社会来说，这一提议的潜在影响是巨大的。

Cardiovascular diseases are both common and deadly. For example, peripheral artery disease affects more than 10M Americans resulting in more than 150,000 limb amputations each year in the U.S. In addition, more than 300,000 patients have coronary artery bypass grafting (surgical revascularization). Current medical therapies for vascular disease include limb amputation and vascular bypass grafting--these therapeutic interventions have significant limitations. These diseases are chronic, debilitating, lethal and they warrant new and novel therapies. Previous studies have demonstrated the essential role for pioneer factors that modulate chromatin accessibility and thereby impact the binding of early transcriptional regulators for lineage specification. We have recently demonstrated that ETV2 is an essential pioneer factor for endothelial, vascular and blood lineages. We have used global and conditional gene disruption strategies, fate-mapping, gene editing, single cell RNA-seq, ATAC-seq and ChIP-seq assays to provide supportive data for this application. In addition, we defined an important ETV2-miR130a-PDGFRa cascade that governs endothelial development. Furthermore, our recent publications and our preliminary data support the overall hypothesis that ETV2 is a pioneer factor that regulates the specification of the endothelial lineage. In these proposed studies, we will use a number of unique genetic models that we have engineered and we take an innovative strategy to define the mechanisms whereby ETV2 functions as a pioneer factor to regulate cardiovascular regeneration. To examine our hypotheses, we will address the following specific aims: Specific Aim #1: Specific Aim #1: To further define the mechanisms whereby ETV2 functions as a pioneer factor during embryogenesis and reprogramming to the endothelial lineage; Specific Aim #2: To define the role of chromatin modifying factors and ETV2 during embryogenesis and reprogramming to the endothelial lineage and Specific Aim #3: To examine the factors that promote ETV2 mediated reprogramming of the endothelial lineage in vitro and in vivo. These aims will utilize our recently engineered genetic mouse models, ATAC-seq, MNase-seq, ChIP-seq, inducible mouse model, cardiac injury model in the adult mouse, novel and bioinformatics algorithms to comprehensively define the mechanisms whereby ETV2 functions as a pioneer factor and will serve as prelude for therapeutic initiatives to engineer and promote regeneration of the cardiovascular lineages. Given the tremendous morbidity and mortality of cardiovascular disease in our society, the potential impact of this proposal is significant.