Metabolomic Profiling of Explanted Pulmonary Arterial Hypertension Lungs

 描述(申请人提供)：肺动脉高压(PAH)是一种致命性疾病，其特征是内皮细胞功能障碍和进行性向内血管生长，导致高血压并最终导致右心衰竭。目前批准的PAH疗法侧重于扩张结构异常的血管以改善生活质量，但肺移植仍然是晚期疾病的唯一治愈方法。PAH与代谢缺陷有关，因为患者表现出胰岛素抵抗和细胞内葡萄糖代谢受损。在这里，我们试图通过描绘被认为是疾病起源的内皮细胞的代谢组，来识别与PAH相关的异常代谢途径作为治疗干预的机会。此外，我们试图了解是否代谢组改变或翻译后修饰状态支持与多环芳烃相关的异常的PPARγ-蛋白质复合体。为了进行研究，我们将从内皮细胞中免疫沉淀PPARγ，并应用代谢组学和蛋白质组学分析来鉴定结合的小分子和共价蛋白质修饰。与多环芳烃相关的改变的代谢物和异常的代谢途径将通过细胞分析中的化学和遗传工具进行验证。这项建议的首要目标是描述PAH的分子基础，以此作为确定新的治疗策略的手段。

 DESCRIPTION (provided by applicant): Pulmonary arterial hypertension (PAH) is a lethal disease characterized by endothelial cell dysfunction and progressive inward vascular growth that leads to hypertension and ultimately right heart failure. Currently approved PAH therapies focus on dilating structurally-abnormal vessels for improved quality of life, however lung transplantation remains the only cure for advanced disease. PAH is associated with metabolic defects as patients demonstrate insulin resistance and impaired intracellular glucose metabolism. Here we seek to identify aberrant metabolic pathways associated with PAH as opportunities for therapeutic intervention, by profiling the metabolome of endothelial cells that have been implicated as the site of disease origin. In addition, we seek to understand whether an altered metabolome or post-translational modification status underlies the aberrant PPARγ-protein complexes associated with PAH. To investigate we will immunoprecipitate PPARγ from endothelial cells and apply metabolomic and proteomic analysis to identify bound small molecules and covalent protein modifications. Altered metabolites and aberrant metabolic pathways associated with PAH will be validated with chemical and genetic tools in cellular assays. The overarching goal of this proposal is to characterize the molecular underpinnings of PAH as a means to identify novel therapeutic strategies.