Endothelial-Interstitial Interactions in Aortic Valve Homeostasis and Disease

项目摘要 近400万65岁以上的美国人患有钙化性主动脉瓣疾病（CAVD）。CAVD发病机制是 胆固醇代谢调节剂无法治疗的活跃生物过程。有 目前没有成功的特异性靶向CAVD的生物靶标或治疗剂。主动脉瓣尖 体内平衡和发病机制是由居民之间复杂和知之甚少的相互作用调节的， 表面瓣膜内皮细胞（VEC）和下面的瓣膜间质细胞（维克）。虽然几乎所有的研究都 专注于维克，它不能从它们周围的VEC中分离出来，并忠实地揭示了 本研究旨在阐明维克、血管内皮细胞及其在CAVD中的作用， 在细胞和分子水平上的相互作用，用于瓣膜稳态和CAVD。我们的初步数据使用新的 瓣膜谱系特异性遗传动物模型显示，NFκB（一种主基因，介导 炎症），或Notch 1（细胞命运和行为的主要调节因子）仅在VEC中不起作用 导致小鼠CAVD。此外，我们创新的3D VEC体外培养鉴定了NFκB活化在VEC中的表达。 血管内皮细胞促进血管内皮细胞向间充质细胞转化，形成新的间充质细胞 钙化基质的祖细胞T-VIC。这些令人兴奋的新结果激发了NFκ B-Notch 1 通过预防或产生这些T-VIC来控制瓣膜稳态和CAVD发病机制。这项建议 将在三个具体目标中进行测试。首先，我们将阐明Notch-NFkB在VEC稳态中的作用， CAVD相关发病机制（目的1）。然后，我们将阐明T-维克如何影响VEC和/或维克表型， 新型CAVD相关疾病中的3D共培养（目的2）。目的3评价NFκ B-Notch 1在肿瘤细胞中的作用 通过减少T-VIC在体内对CAVD起始和进展的信号再平衡。完成这一 该项目将产生关于主动脉瓣稳态的细胞间调节的重要信息， CAVD的发病过程。通过揭示参与VEC和维克的独特表型特征， 我们的研究还将确定和激发新的分子候选人，专门针对细胞特异性 CAVD过程的特征，以获得潜在的诊断和治疗益处。

Project Summary Nearly 4 million Americans over 65 are living with calcific aortic valve disease (CAVD). CAVD pathogenesis is an active biological process that is untreatable by cholesterol metabolism modifying agents. There are currently no successful biological targets or therapeutic agents that specifically target CAVD. Aortic valve cusp homeostasis and pathogenesis is regulated by complex and poorly understood interactions between resident surface valve endothelial cells (VEC) and underlying valve interstitial cell (VIC). While almost all research has focused on VIC, which cannot be isolated from VEC that surround them and faithfully reveal the biology of aortic valves and pathobiology of CAVD, this research project aims to unravel the roles of VIC, VEC and their interactions, at cell and molecule levels, for valve homeostasis and CAVD. Our preliminary data using new valve lineage specific genetic animal models show that activation of NFκB (a master gene mediating inflammation) in VEC and VIC, or inaction of Notch1 (a major regulator of cell fate and behavior) in VEC only results in CAVD in mice. Further, our innovative 3D in vitro culture of VEC identifies that NFκB activation in VEC promotes VEC undergo endothelial-to-mesenchymal transformation and generate a novel mesenchymal progenitor T-VIC that calcifies matrix. These exciting new results motivate the hypothesis that NFκB-Notch1 control valve homeostasis and CAVD pathogenesis via prevention or generation of these T-VIC. This proposal will be tested in three Specific Aims. First we will elucidate the role of Notch-NFkB in VEC homeostasis and CAVD related pathogenesis (Aim 1). Then we will elucidate how T-VIC affect VEC and/or VIC phenotypes in 3D co-culture in novel CAVD-related conditions (Aim 2). Aim 3 will evaluate the efficacy of NFκB-Notch1 signaling rebalancing on CAVD initiation and progression in vivo via reduction of T-VIC. The completion of this project will generate significant information regarding intercellular regulation of aortic valve homeostasis and the CAVD pathogenic process. By revealing the unique phenotypic signatures of participation VEC and VIC, our study will also identify and motivated new molecular candidates that specifically target the cell specific characteristics of the CAVD process for potential diagnostic and therapeutic benefit.