Endothelial mechanotransduction and metabolic remodeling

内皮力转导和代谢重塑

基本信息

批准号：
10468115
负责人：
JEFFREY R FINEMAN
金额：
$ 39.94万
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/10468115
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 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 Lead Left Lesion Ligation Liquid substance Live Birth Lung Mediating Mediator of activation protein Medical Metabolic Metabolism Mitochondria Modeling Morbidity - disease rate Natural History Operative Surgical Procedures Pathway interactions Patients Pattern Pentosephosphate Pathway Periodicity Phenotype Plasma Play Production Pulmonary artery structure Regulation Role Severities Severity of illness Shunt Device Signal Pathway Signal Transduction Source Specificity Stimulus Stretching Testing Therapeutic Intervention Validation base 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.

项目摘要肺血管疾病（PVD）是先天患者发病率和死亡率的重要来源心脏病（CHD）。这些患者的PVD的自然病史揭示了重要的病理生理与异常的肺血流（PBF）和压力相关的差异。心脏缺陷的患者从全身性的直接压力刺激中，暴露于肺脉管系统会增加流量心室发展PVD比出现缺陷的患者更大的事件和严重程度，导致PBF增加独自的。肺内皮细胞（EC）是疾病的积分介体，因为它们暴露于这些正常和血液动力学异常（机械）力，包括剪切应力，静水压力和环状应变。我们的实验室在胎儿羔羊中开发了两种不同的临床相关模型：（1）左肺主要导致右肺PBF增加的动脉（LPA）结扎；（2）主动脉肺位置会导致PBF和压力增加。我们的初步数据表明，在4-6周年龄，模型羔羊在内皮细胞信号传导和血管功能中表现出明显的畸变。例如，从每个羔羊模型对原代肺动脉内皮细胞（PAEC）进行的RNASEQ分析证明了明显不同的基因表达模式，在孤立血管中的研究表明不同血管反应性的改变。此外，我们在体内和体外数据中生成了新颖的数据，证明了这一点生物力学力的附加作用 - 流体剪切应力和压力引起的环状拉伸 - 是因为导致内皮功能障碍（ENOS解偶联），代谢的细胞信号传导途径的扰动重新编程（ROS驱动的HIF-1A和C-MYC激活），以及一种超增殖的，抗凋亡，内皮细胞表型。基于这些数据，我们将在项目＃1中检验的总体假设是不同与PBF增加和压力相比，与PBF增加相关的机械力，衍生图案基因表达和血管功能的改变，这是PVD入射和进展的基础与CHD相关。特别是，我们假设通过ATP - 依赖性的HSP90活性和C-MYC介导的谷氨酰胺孤立性。添加压力感应循环然而，伸展会导致HIF-1α通过增加而驱动Warburg代谢和EC过度增殖线粒体（MT） - ROS产生，但以ATP依赖性HSP90活性为代价，没有信号传导。这总体假设将以三个相关但独立的特定目的进行检验。作为当前的PVD处理方法基于疾病的严重程度，而不是潜在的病理生物学，成功完成拟议的研究可能会导致针对PVD 2°的靶向治疗方法，并告知其他 PVD的类型，其中异常的机械力参与疾病进展。