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的发病机制是 胆固醇代谢调节剂无法治疗的一种活跃的生物过程。确实有 目前还没有成功的生物靶点或专门针对CAVD的治疗剂。主动脉瓣瓣尖 动态平衡和发病机制由居民之间复杂而鲜为人知的相互作用来调节 表面瓣膜内皮细胞(VEC)和下层瓣膜间质细胞(VIC)。虽然几乎所有的研究都有 聚焦于VIC，它不能从周围的VEC中分离出来，并忠实地揭示了 本研究旨在揭示血管内皮细胞、血管内皮细胞及其受体在心血管疾病中的作用。 在细胞和分子水平上的相互作用，瓣膜动态平衡和CAVD。我们的初步数据使用新的 瓣膜谱系特异的遗传动物模型显示，NFκB(一种主要调节基因)的激活 炎症)，或仅在血管内皮细胞中Notch1(细胞命运和行为的主要调节因子)不起作用 结果在小鼠中出现CAVD。此外，我们创新的血管内皮细胞3D体外培养发现，NFκB在血管内皮细胞中的激活 血管内皮细胞促进内皮细胞向间充质转化并生成新的间质 使基质钙化的祖细胞T-VIC。这些令人兴奋的新结果支持了一种假设，即NFκB-Notch1 通过预防或产生这些T-VIC来控制瓣膜动态平衡和CAVD的发病。这项建议 将在三个具体目标上进行测试。首先，我们将阐明Notch-NFkB在血管内皮细胞动态平衡和 CAVD相关发病机制(目标1)。然后我们将阐明T-VIC是如何影响VEC和/或VIC表型的 在新的CAVD相关条件下3D共培养(目标2)。目的3将评估核因子κB-Notch1的疗效 通过降低T-VIC在体内CAVD启动和进展中的信号再平衡。这项工程的完成 该项目将产生关于主动脉瓣内稳态的细胞间调节和 CAVD的致病过程。通过揭示参与VEC和VIC的独特表型特征， 我们的研究还将确定和激励新的候选分子，这些候选分子专门针对特定的细胞 CAVD过程的特点对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.