Roles of neuropilin-1 in endothelial cell dysfunction

项目摘要/摘要 射血分数保留的心力衰竭(HFpEF)影响着300多万美国成年人 而且没有有效的治疗方法。改善内皮细胞(EC)的一般策略的可喜效果 HFpEF患者的功能已经强调了EC功能障碍在疾病发病机制中的关键作用。 然而，EC功能障碍的分子机制及其在HFpEF发病机制中的作用 人们对此仍然知之甚少。最近的一项患者研究发现，血清神经毛细蛋白-1(Nrp1)是一种预后标志物 在HFpEF患者中。我们的初步数据表明，内皮细胞Nrp1在心脏组织中增加 小鼠HFpEF模型和内皮Nrp1缺乏的小鼠显示EC和心脏舒张期的改善 在这些模型中的功能。肿瘤坏死因子-α(肿瘤坏死因子-α)是血管内皮细胞功能障碍的主要诱因 与HFpEF的疾病进展有关。我们的初步数据表明，Nrp1基因的敲除 降低肿瘤坏死因子α诱导的黏附分子表达，升高内皮一氧化氮水平 内皮细胞中的合酶(ENOS)。计算对接模型和蛋白质结合分析共同表明 Nrp1与肿瘤坏死因子α的直接相互作用。基于这些结果，该项目将检验中心假设 内皮Nrp1作为一种与TNFR共同作用的新受体，通过以下途径导致HFpEF的发病 增加心脏炎症和损害心肌一氧化氮的生物利用度。 提出了两个具体的目标：目标1：描述在EC中Nrp1和肿瘤坏死因子α/肿瘤坏死因子受体之间的串扰 功能障碍。我们将使用微板法研究Nrp1与肿瘤坏死因子α/肿瘤坏死因子受体之间的相互作用 结合实验和培养的内皮细胞。Nrp1在肿瘤坏死因子α诱导的EC功能障碍中的作用将在两个 小鼠和斑马鱼内皮细胞特异性Nrp1基因敲除模型。目的2：揭示内皮细胞的作用 Nrp1在HFpEF发病机制中的作用我们将使用小鼠HFpEF模型来研究内皮Nrp1如何 与TNFR1协同控制HFpEF的疾病发生和发展。

PROJECT SUMMARY/ABSTRACT Heart failure with preserved ejection fraction (HFpEF) affects more than 3 million adults of the United States and has no effective therapy. The promising effect of general strategies for improving endothelial cell (EC) function in HFpEF patients has highlighted the crucial roles of EC dysfunction in the disease pathogenesis. However, the molecular mechanism of EC dysfunction and how it contributes to the pathogenesis of HFpEF are still poorly understood. A recent patient study identified serum neuropilin-1 (NRP1) as a prognostic marker in HFpEF patients. Our preliminary data indicate that endothelial NRP1 is increased in the cardiac tissues of murine HFpEF models and mice deficient of endothelial NRP1 show improved EC and cardiac diastolic functions in these models. Tumor necrosis factor-α (TNFα) is a major inducer of EC dysfunction and associates with the disease progression of HFpEF. Our preliminary data suggest that knockdown of NRP1 reduces TNFα-induced expression of adhesion molecules but enhances the levels of endothelial nitric oxide synthase (eNOS) in ECs. Computational docking model and protein binding assay collectively indicate the direct interaction between NRP1 and TNFα. Based on these results, this project will test the central hypothesis that endothelial NRP1, acting as a novel co-receptor with TNFR, leads to the pathogenesis of HFpEF by increasing cardiac inflammation and impairing myocardial nitric oxide bioavailability. Two Specific Aims are proposed: Aim 1: Delineate the crosstalk between NRP1 and TNFα/TNFR in EC dysfunction. We will characterize the interaction between NRP1 and TNFα/TNFR using microplate-based binding assay and cultured ECs. The role of NRP1 in TNFα-induced EC dysfunction will be examined in both mouse and zebrafish endothelial cell specific NRP1 knockout models. Aim 2: Reveal the role of endothelial NRP1 in the pathogenesis of HFpEF. We will use mouse HFpEF models to investigate how endothelial NRP1 cooperates with TNFR1 to control the disease initiation and progression of HFpEF.