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

肺动脉高压细胞与血管基质之间的动态互易性评估

• 批准号: 10686936
• 负责人: Rubin M. Tuder
• 金额: $ 43.79万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10686936
• 关键词: Address; Animal Diseases; Animal Model; Blood Vessels; Cattle; Cells; Complement; Complement Activation; Complement Inactivators; Data; Data Correlations; Disease model; Elements; Enhancers; Environment; Evaluation; Extracellular Matrix; Fingerprint; Gene Expression; Genetic Transcription; Genomic approach; Genomics; Goals; Heterogeneity; Human; Hypertrophy; Image; Immune; Immunofluorescence Immunologic; Individual; Inflammation; Inflammatory; Intervention; Investigation; Lesion; Link; Lung; Lung diseases; Macrophage; Mediating; Messenger RNA; Methods; Modeling; Molecular; Molecular Profiling; Patients; Phenotype; Plasma; Plasma Proteins; Proteins; Proteome; Proteomics; Pulmonary Hypertension; Pulmonary arterial remodeling; Rattus; Role; SU 5416; Sampling; Serum; Structure of parenchyma of lung; Testing; Therapeutic; Tissues; cohort; complement system; disease heterogeneity; extracellular; genetic element; hemodynamics; human disease; human tissue; hypoxia-induced pulmonary hypertension; inflammatory marker; inflammatory milieu; member; mouse model; novel strategies; pre-clinical; primary pulmonary hypertension; pulmonary arterial hypertension; pulmonary vascular disorder; pulmonary vascular remodeling; synergism; transcriptome

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.

越来越多的证据表明，炎症是肺血管的触发剂和增强剂。 肺动脉高压(PH)的重构。本PPG侧重于补体作为触发因素的作用 肺血管疾病与肺高压。项目3的总体指导思想是疾病的关键角度 多个水平的异质性：在整个人类病变肺水平，在病变肺的病变水平，在 IPAH肺和PH动物模型之间的相似病变水平。这项提议的中心目标是 说明在项目1、2和4中开展的力学研究的意义。 我们的中心假设是，包括RNA表达和细胞在内的关键分子指纹 细胞外蛋白质组可用于确定肺血管损伤的分子特异性 啊哈。这些特征将反映补体的长期激活和随后的重塑 血管周围环境。我们依靠一种基于对人类疾病的审问的综合方法 和控制肺，以及一大群特征良好的IPAH患者和对照组的血浆(目标1)，稳健 重度肺高压模型(目标2)，以及牛缺氧性肺高压模型的机制研究(目标3)，使用 高通量的基因组学、蛋白质组学和肺组织水平的免疫定位方法。 具体地说，AIM 1将检验这一假设，即在IPAH肺中存在补体激活的证据 血管损伤，与特定的蛋白质组和基因组指纹相关；AIM 2将识别共享 从人IPAH建立的分子特征与牛和大鼠SU5416模型的比较 PH，重点是针对补体和细胞外基质的治疗策略的优先顺序 以及，AIM 3将在一个大的PH动物模型中测试补体阻断将 导致促炎微环境减少，肺血管蛋白质组正常化。 这一建议具有非常重要的意义，因为它解决了肺间质细胞分子异质性的关键方面 血管损伤以及患者和动物模型；它提供了一个中心平移角度 项目1、2和4中的机制研究，因为它试图确定疾病患者的补体激活标志物 人体组织和强健的疾病模型。此外，它还将提供证据，将补体封锁与 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， 包括在皮损水平上评估蛋白质组和转录组，高通量 组织水平的免疫荧光成像，以及阻断补体的新方法。