Toward Therapeutic Manipulation of Endothelial to Mesenchymal Transition

内皮细胞向间质细胞转化的治疗性调控

• 批准号: 9208153
• 负责人: Jason Ciril Kovacic
• 金额: $ 42.38万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-25 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9208153
• 关键词: Adult; Animal Model; Animals; Apolipoprotein E; Arteries; Arteriovenous fistula; Atherosclerosis; Autopsy; Blood Vessels; Cardiac development; Cardiovascular Diseases; Cardiovascular system; Cause of Death; Cell Line; Cell physiology; Cessation of life; ChIP-seq; Clinical; Code; Collagen; Coronary; Coronary Arteriosclerosis; DNA; Data; Data Set; Disease; Embryonic Development; Endothelial Cells; Epidemic; Epigenetic Process; Excipients; Family suidae; Fibrosis; Fistula; Gene Expression; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Genetic study; Genotype; Histones; Human; In Vitro; Individual; Inflammation; Inherited; Knock-out; Knowledge; Mediating; Mesenchymal; Modeling; Molecular; Mus; Operative Surgical Procedures; Organ Transplantation; Oxidative Stress; Patients; Persons; Phenotype; Play; Prevalence; Process; Production; Quantitative Trait Loci; RNA; RNA Sequences; Regulatory Pathway; Research; Rodent; Role; Sampling; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; System; Therapeutic; Tissues; Transforming Growth Factor beta; Untranslated RNA; Vascular Graft; Vascular remodeling; Vein graft; Veins; Virus; adeno-associated viral vector; clinical translation; coronary fibrosis; disorder risk; epithelial to mesenchymal transition; healing; histone modification; improved; in vivo; infancy; knock-down; mortality; network models; novel; novel therapeutics; pre-clinical; pressure; public health relevance; response to injury; risk variant; vector; vector control

 DESCRIPTION (provided by applicant): Endothelial to Mesenchymal Transition (EndMT) plays a key role in embryonic development and in adult cardiovascular disease (CVD). While our team is at the forefront in defining the importance of EndMT in CVD, our overall knowledge of this process remains in its infancy. The broad objective of this R01 is to leverage novel discoveries made by our group to define the core mechanisms of EndMT in adult CVD, with an ultimate view to clinical translation. Aim 1 will define the causal role of Zeb2 in EndMT and atherosclerotic disease. We have identified a likely biologic intersection point between CAD, EndMT and the gene Zeb2. Zeb2 is known to modulate EndMT-related cellular processes, while a Zeb2-associated polymorphism (rs2252641) is one of only 46 independent CAD risk alleles. We further validated the importance of Zeb2 in CAD using our unique STARNET dataset, a genetics-of-gene expression study involving 1062 patients with and without CAD. In Aim 1 we will combine in vitro studies of Zeb2 knockdown during EndMT, in vivo rodent studies of tissue- specific Zeb2 knockdown in an atherosclerosis model, and systems genetics studies of Zeb2 using the STARNET dataset. We anticipate that Aim 1 will make major inroads on our mechanistic understanding of Zeb2, its effect in promoting CAD and its role in EndMT. Given that Zeb2 is a CAD risk allele but yet its function in this disease is unknown, this Aim is of significant clinical importance. Aim 2 will define the effect of EndMT inhibition in a large animal model of vein graft remodeling and obtain preclinical proof-of- principle data for this approach. We have shown that EndMT is critical during vein graft remodeling when sections of vein are exposed to arterial pressure. Antagonism of endothelial TGF-β signaling in mice by altered Smad2/3-Snai2 signaling led to reduced EndMT during vein graft remodeling and less neointimal formation. In Aim 2 we will generate adeno-associated viral vector (AAV) 1 constructs targeting Smad3 and Snai2 and apply these to an established arterio-venous fistula porcine model to define the effect of EndMT inhibition on vein graft remodeling. We expect Aim 2 to provide definitive data as to the possibility of inhibiting EndMT to improve vein graft remodeling such that grafts/fistulae have reduced neointima, increased luminal size and ultimately, improved patency. Aim 3 will define key drivers and regulators of EndMT in the adult. Using our unique in vitro EndMT model we will drive disease-relevant endothelial cells toward a mesenchymal phenotype and conduct in-depth studies of the transcriptional and epigenetic regulators of EndMT. Systems genetics will be used to integrate these transcriptional, epigenetic and other regulators of EndMT to define key hierarchical drivers of EndMT. We expect that Aim 3 will greatly enrich our understanding of the core molecular and epigenetic mechanisms of EndMT. Collectively, this project will shift current paradigms and firmly establish the role of EndMT, and the therapeutic opportunities for EndMT manipulation, in adult CVD.

 描述（应用程序提供）：间充质转变（ENDMT）的内皮在胚胎发育和成人心血管疾病（CVD）中起关键作用。尽管我们的团队在确定CVD中EndMT的重要性方面处于最前沿，但我们对这一过程的总体知识仍处于起步阶段。该R01的广泛目标是利用我们小组的新发现，以定义成人CVD中EndMT的核心机制，并最终观察到临床翻译。 AIM 1将定义Zeb2在EndMT和动脉粥样硬化疾病中的因果作用。我们已经确定了CAD，EndMT和Gene Zeb2之间的生物相交点。已知ZEB2调节与EndMT相关的细胞过程，而ZEB2相关的多态性（RS2252641）是仅有的46个独立的CAD风险等位基因之一。我们使用独特的Starnet数据集进一步验证了Zeb2在CAD中的重要性，这是一项遗传学表达研究，涉及1062名患有和不具有CAD的患者。在AIM 1中，我们将在EndMT期间对Zeb2敲低的体外研究结合，在动脉粥样硬化模型中对组织 - 特异性Zeb2敲低的体内啮齿动物研究，以及使用Starnet数据集对ZEB2进行的系统遗传学研究。我们预计AIM 1将对我们对Zeb2的机械理解，其在促进CAD及其在EndMT中的作用的影响大大介绍。鉴于Zeb2是CAD风险等位基因，但其在该疾病中的功能尚不清楚，因此该目标具有重要的临床重要性。 AIM 2将定义大型动物中EndMT抑制作用的效果 静脉谷物重塑的模型并获得此方法的临床前原理数据。我们已经表明，当静脉截面暴露于人工体压力时，EndMT至关重要。改变SMAD2/3-SNAI2信号在小鼠中内皮TGF-β信号传导的拮抗作用导致静脉晶粒重塑过程中的末端MT降低，形成较少。在AIM 2中，我们将生成靶向SMAD3和SNAI2的腺相关病毒载体（AAV）1构建体，并将其应用于已建立的动脉疫苗猪猪猪模型，以定义EndMT抑制对静脉谷物重塑的影响。我们期望AIM 2提供定义的数据，以抑制EndMT改善静脉谷物重塑的可能性，从而使移植物/瘘管减少了新内膜，增加了腔尺寸并最终提高了通畅性。 AIM 3将定义成人EndMT的关键驱动因素和监管机构。使用我们独特的体外ENDMT模型，我们将驱动与疾病相关的内皮细胞朝着间质表型进行，并对ENDMT的转录和表观遗传调节剂进行深入研究。系统遗传学将用于整合EndMT的这些转录，表观遗传和其他调节剂，以定义EndMT的关键层次驱动器。我们预计目标3将极大地丰富我们对EndMT的核心分子和表观遗传机制的理解。总的来说，该项目将改变当前的范例，并首先确立EndMT的作用，以及在成人CVD中进行EndMT操纵的治疗机会。