Evaluation of the Dynamic Reciprocity Between Cells and the Vessel Matrix in Pulmonary Hypertension

越来越多的证据表明，炎症是肺血管病变的触发因素和增强因素， 肺动脉高压（PH）的重塑。该PPG侧重于补体作为免疫调节剂的触发剂的作用。 肺血管疾病和PH。项目3的总体指导思想是疾病的关键角度 在多个水平的异质性：在整个患病的人肺水平，在患病肺中的病变水平，在 IPAH肺和PH动物模型之间相似病变的水平。本提案的中心目标是 提供项目1、2和4中开展的机理研究的意义。 我们的中心假设是，关键的分子指纹，包括RNA表达和细胞组合， 与细胞外蛋白质组可用于定义分子特异性肺血管病变， 呸。这些特征将反映出补体的长期激活和随后的重塑 血管周围环境我们依靠的是一种基于对人类疾病的审讯的综合方法， 和对照肺，以及大量充分表征的IPAH患者和对照的血浆（目的1），稳健 严重PH模型（目标2），以及缺氧PH牛模型（目标3）的机制研究，使用 肺组织水平的基因组学、蛋白质组学和免疫定位的高通量方法。 具体而言，AIM 1将检验以下假设：IPAH肺内存在补体激活的证据。 血管病变，与特定的蛋白质组学和基因组指纹相关; AIM 2将识别共享的 与牛和大鼠SU 5416模型相比， PH，重点关注靶向补体和细胞外基质的治疗策略的优先顺序 并且，AIM 3将检验以下假设：在PH的大型动物模型中， 导致促炎微环境降低和肺血管蛋白质组正常化。 这一建议是非常重要的，因为它解决了肺组织中分子异质性的关键方面。 血管病变和患者之间以及动物模型;其用于提供中心平移角， 项目1、2和4中的机制研究，因为它试图确定疾病中的补体激活标志物， 人体组织和健全的疾病模型。此外，它将提供证据，证明补体阻断与 逆转PH促炎表型和PH蛋白质组。项目3引入了非常新颖的方法， 包括在病变水平上的蛋白质组和转录组的评估， 组织水平的免疫荧光成像，以及阻断补体的新方法。

There is growing evidence of the role of inflammation as a trigger and enhancer of pulmonary vascular remodeling in pulmonary hypertension (PH). This PPG focuses on the role of complement as a trigger of pulmonary vascular disease and PH. The overall guiding concept for Project 3 is the key angle of disease heterogeneity at multiple levels: at the whole diseased human lung level, lesional level in a diseased lung, at the level of similar lesions among IPAH lungs, and animal models of PH. It is the central goal of this Proposal to provide the significance of the mechanistic studies developed in Projects 1,2, and 4. Our central hypothesis is that key molecular fingerprints including RNA expression and cellular combined with extracellular proteomes can be used to define molecularly specific pulmonary vascular lesions in PAH. These profiles will reflect prolonged activation of complement and the subsequent remodeled perivascular environment. We rely on a comprehensive approach based on interrogation of human diseased and control lungs, and plasma of a large cohort of well characterized IPAH patients and controls (Aim 1), robust models of severe PH (Aim 2), and mechanistic investigations in the bovine model of hypoxic PH (Aim 3), using high throughput approaches of genomics, proteomics, and immune localization at the lung tissue level. Specifically, AIM 1 will test the hypothesis that there is evidence of complement activation in IPAH pulmonary vascular lesions, which are related to specific proteomic and genomic fingerprints; AIM 2 will Identify shared molecular signatures established from the human IPAH as compared with the bovine and rat SU5416 models of PH, with a focus on prioritization of therapeutic strategies targeting complement and extracellular matrix remodeling; and, AIM 3 will test the hypothesis that blockade of complement in a large animal model of PH will lead to decreased proinflammatory microenvironment and normalization of the pulmonary vascular proteome. This proposal is highly significant as it addresses the key aspect of molecular heterogeneity among pulmonary vascular lesions and among patients, and animal models; it serves to provide a central translational angle to mechanistic studies in Projects 1,2, and 4 as it seeks to identify complement activation markers in diseased human tissue and robust models of disease. Moreover, it will provide evidence linking complement blockade in reverting a PH proinflammatory phenotype and PH proteome. Project 3 introduces highly novel approaches, including the assessment of the proteome and transcriptome at the lesional level, high throughput immunofluorescence imaging at the tissue level, and novel approaches to block complement.