Endothelial mechanotransduction and metabolic remodeling

内皮力转导和代谢重塑

基本信息

批准号：
10705691
负责人：
JEFFREY R FINEMAN
金额：
$ 40.32万
依托单位：
FLORIDA INTERNATIONAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-20 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10705691
关键词：
Age Animals Apoptotic Attention Attenuated Bioenergetics Biomechanics Blood Pressure Blood Vessels Blood flow Blood specimen Cardiopulmonary Catabolism Child Chronic Congenital Heart Defects Data Defect Development Disease Disease Progression Disparate Endothelial Cells Endothelium Enzymes Exposure to Fetal Sheep Gene Expression Gene Expression Profile Glutamine Glycolysis Growth Heart Abnormalities Hexokinase 2 Human Hydrostatic Pressure In Vitro Incidence Intervention Laboratories Left Lesion Ligation Liquid substance Live Birth Lung Mechanics Mediating Mediator Medical Metabolic Metabolism Mitochondria Modeling Morbidity - disease rate Natural History Operative Surgical Procedures Pathway interactions Patients Pattern Pentosephosphate Pathway Periodicity Phenotype Plasma Play Production Proliferating Pulmonary artery structure Regulation Role Severities Severity of illness Shunt Device Signal Pathway Signal Transduction Source Specificity Stimulus Stretching Testing Therapeutic Intervention Validation biomarker discovery bromopyruvate c-myc Genes clinically relevant congenital heart disorder design endothelial dysfunction hemodynamics improved in vivo inhibitor lamb model lipid biosynthesis mechanical force mechanotransduction metabolomics mortality nitration novel oxidation postnatal preservation pressure pulmonary artery endothelial cell pulmonary vascular disorder pulmonary vascular remodeling screening shear stress targeted treatment transcriptome sequencing vascular abnormality

项目摘要

PROJECT SUMMARY Pulmonary vascular disease (PVD) is an important source of morbidity and mortality in patients with congenital heart disease (CHD). The natural history of PVD in these patients reveals the important pathophysiologic differences associated with abnormal pulmonary blood flow (PBF) and pressure. Patients with cardiac defects that expose the pulmonary vasculature to increased flow with a direct pressure stimulus from the systemic ventricle develop PVD with greater incidence and severity than patients with defects resulting in increased PBF alone. Pulmonary endothelial cells (EC) are integral mediators of disease, due to their exposure to these normal and abnormal hemodynamic (mechanical) forces including shear stress, hydrostatic pressure, and cyclic strain. Our laboratory has developed two distinct, clinically relevant models of CHD in fetal lambs: (1) left pulmonary artery (LPA) ligation that primarily results in increased PBF to the right lung; and (2) aortopulmonary shunt placement that results in increased PBF and pressure. Our preliminary data demonstrate that at 4-6 weeks of age, model lambs manifest distinct aberrations in endothelial cell signaling and vascular function. For example, RNAseq analysis performed on primary pulmonary artery endothelial cells (PAEC) from each lamb model demonstrates markedly distinct gene expression patterns, and studies in isolated vessels demonstrate disparate alterations in vascular reactivity. Moreover, we have generated novel in vivo and in vitro data demonstrating that the additive effects of the biomechanical forces—fluid shear stress and pressure induced cyclic stretch—cause perturbations in cellular signaling pathways that result in endothelial dysfunction (eNOS uncoupling), metabolic reprogramming (ROS driven HIF-1a, and c-MYC activation), and a hyper-proliferative, anti-apoptotic, endothelial cell phenotype. Based on these data, the overall hypothesis we will test in Project #1, is that the distinct mechanical forces associated with increased PBF compared to increased PBF and pressure, induce patterned alterations in gene expression and vascular function that underlie the incidence and progression of PVD associated with CHD. Specifically, we hypothesize that flow-alone maintains NO signaling through ATP- dependent hsp90 activity and c-MYC-mediated glutamine anaplerosis. The addition of pressure induced cyclic stretch, however, leads to HIF-1α driven Warburg metabolism and EC hyper-proliferation via increases in mitochondrial (mt)-ROS production, but at the expense of ATP-dependent hsp90 activity and NO signaling. This overall hypothesis will be tested in three inter-related, but independent, Specific Aims. As current PVD treatment approaches are based on disease severity as opposed to underlying pathobiology, the successful completion of the proposed studies may lead to targeted therapeutic approaches for PVD 2° to CHD, as well as inform other types of PVD, in which abnormal mechanical forces participate in disease progression.

项目概要肺血管疾病（PVD）是先天性肺病患者发病和死亡的重要来源这些患者的PVD自然史揭示了重要的病理生理学特征。与心脏缺陷患者的肺血流量（PBF）和压力异常相关的差异。使肺血管系统暴露在来自全身系统的直接压力刺激下增加的流量与导致 PBF 增加的缺陷患者相比，心室发生 PVD 的发生率和严重程度更高由于肺内皮细胞（EC）暴露于这些正常细胞，因此它们是疾病的重要介质。以及异常的血流动力学（机械）力，包括剪切应力、静水压力和循环应变。我们的实验室开发了两种不同的、临床相关的胎羔 CHD 模型：(1) 左肺动脉 (LPA) 结扎，主要导致右肺 PBF 增加；以及 (2) 主动脉肺分流我们的初步数据表明，在 4-6 周时，会导致 PBF 和压力增加。随着年龄的增长，模型羔羊的内皮细胞信号传导和血管功能表现出明显的异常。对每只羔羊模型的原代肺动脉内皮细胞 (PAEC) 进行 RNAseq 分析表现出明显不同的基因表达模式，并且在分离的血管中的研究表明不同的基因表达模式此外，我们已经生成了新的体内和体外数据，证明了这一点。生物力学力（流体剪切应力和压力引起的循环拉伸）的累加效应细胞信号通路的扰动导致内皮功能障碍（eNOS 解偶联）、代谢重编程（ROS 驱动的 HIF-1a 和 c-MYC 激活），以及过度增殖、抗凋亡、内皮细胞基于这些数据，我们将在项目 #1 中测试的总体假设是，不同的细胞表型。与增加的 PBF 和压力相比，与增加的 PBF 相关的机械力会引起图案化 PVD 发生和进展的基因表达和血管功能的改变具体来说，我们勇敢地说，单独的流量通过 ATP-维持 NO 信号传导。依赖的 hsp90 活性和 c-MYC 介导的谷氨酰胺回补增加压力诱导的循环。然而，拉伸会导致 HIF-1α 驱动的 Warburg 代谢和 EC 过度增殖（通过增加线粒体 (mt)-ROS 产生，但以牺牲 ATP 依赖性 hsp90 活性和 NO 信号传导为代价。总体假设将在三个相互关联但独立的具体目标中进行检验，如当前的 PVD 治疗。方法基于疾病的严重程度而不是基础病理学，成功完成拟议的研究可能会导致 PVD 2° 至 CHD 的靶向治疗方法，并为其他疾病提供信息 PVD 类型，其中异常机械力参与疾病进展。