Circulating biomarkers of IPAH.

描述(申请人提供)：儿童或成人的肺动脉高压(PAH)是一种进行性和致命性的疾病，其特征是原因不明的持续的肺动脉压升高。血管扩张剂是治疗的主要药物。然而，20%-30%的患者对血管扩张剂没有反应，无反应者预后较差，最终需要肺移植。由于病理生物学尚不清楚，血管扩张剂治疗的发病率和费用都很高，而且缺乏特异的诊断/预后生物标记物，我们的长期目标是确定IPAH的循环、诊断和预后标记物，以改善长期疗效。为了追求这一目标，我们优化了一种对肺动脉高压患者进行无偏见、深入研究的蛋白质组学方法。在我们使用IPAH儿童纵向血浆样本的初步数据中，我们已经发现IPAH患者的许多血浆蛋白发生了显着变化。因此，我们假设IPAH患者的血浆样本将包含诊断/预后肺和内皮细胞特异性生物标记物。为了验证这一假设，我们将使用一种无偏见的蛋白质组学方法，从肺组织，特别是肺动脉内皮细胞(PAEC)中鉴定独特的循环PAH生物标志物蛋白。目的1.利用最先进的定量生物标记物开发流程，我们将通过比较PHBI库患者(n=10)或APAH(n=10)先天性心脏病患者和(n=10)正常非移植供者(n=10)的匹配血浆、肺和PAEC的蛋白质组，来鉴定循环肺，特别是IPAH的PAEC特异性生物标记物。在目标2中，我们将使用高通量的基于MS的多重定量反应监测(MRM)分析来验证候选的血浆IPAH生物标志物，使用剩余的PHBI库来验证其蛋白质识别，它在儿童和成人中的有效性，并确定其对IPAH和非IPAH的特异性。创新之处在于，这项提案将一种最先进的、无偏见的蛋白质组学发现流程应用于一组新的匹配的PAH和正常血液、肺和PAEC，以识别诊断/预后的IPAH生物标记物。这些方法将首次为IPAH和可能的PAH提供特定的肺生物标记物，为未来的临床试验和新治疗靶点的开发提供丰富的新数据。重要的是，尽管在IPAH的治疗和生存方面已经取得了进展，但其病因仍然很大程度上是未知的。此外，我们缺乏诊断/预测手段，无法轻松、准确地识别对治疗无反应的患者。因此，生物标记物可以填补这一空白，提供降低治疗发病率的手段，适当升级治疗，并作为新疗法的基准措施。 公共卫生相关性：儿童或成人的肺动脉高压(PAH)是一种进行性和致命性的疾病，尽管在IPAH的治疗和生存方面取得了进展，但其病因仍很大程度上仍不清楚。在这项提案中，我们将使用最先进的蛋白质组学流水线来识别和验证特定的肺/肺动脉内皮细胞预后/诊断IPAH生物标记物。因此，生物标记物可以填补这一空白，提供降低治疗发病率的手段，适当升级治疗，并作为新疗法的基准措施。(摘要结束)。

DESCRIPTION (provided by applicant): Pulmonary artery hypertension in children or adults (PAH) is a progressive and fatal disease characterized by sustained elevations of pulmonary artery pressure of unknown etiology. Vasodilators are the mainstay of therapy. However 20-30% of patients do not respond to vasodilators and non-responders have a poor prognosis eventually requiring lung transplantation. As the pathobiology is unknown, vasodilator therapy has significant morbidity and cost and specific diagnostic/prognostic biomarkers are lacking, our long-term goal is to identify circulating, diagnostic and prognostic markers of IPAH to improve long term outcomes. To pursue this goal, we have optimized a non-biased, in-depth proteomics approach in patients with pulmonary hypertension. In our preliminary data using longitudinal plasma samples of children with IPAH we have already identified significant changes in a number of plasma proteins in patients with IPAH. Therefore, we hypothesize that plasma samples from patients with IPAH will contain diagnostic/prognostic lung and endothelial cell specific biomarkers. To examine this hypothesis we will use a non-biased proteomics approach to identify unique circulating PAH biomarker proteins from the lung and specifically pulmonary arterial endothelial cells (PAEC). In Aim 1. Using a state of the art quantitative biomarker development pipeline, we will identify circulating lung and in particular, PAEC specific biomarkers of IPAH by comparing proteomes of matched plasma, lung and PAEC harvested from PHBI Repository patients transplanted for IPAH (n=10) or APAH (n=10) related to congenital heart disease and (n=10) and normal non-transplanted donors (n=10) as controls. In Aim 2. we will verify candidate plasma IPAH biomarkers using high throughput quantitative multiplex MS-based (multple reaction montioring (MRM) assays using the remaining PHBI Repository of plasma samples to verify the protein identification, it's efficacy in both children and adults and determine it's specificity for IPAH vs non-IPAH. The innovation is that this proposal applies a state of the art unbiased proteomics discovery pipeline to a novel set of matched PAH and normal blood, lung and PAEC to identify diagnostic/prognostic IPAH biomarkers. These methods will allow for the first time the potential for specific lung biomarkers of IPAH and possibly PAH in general providing a wealth of new data for future clinical trials and development of new therapeutic targets. The significance is that although there have been advances in therapy and survival with IPAH, the etiology is still largely unknown. In addition we lack the diagnostic/prognostic means to easily and accurately identify patients that are unresponsive to therapy. Therefore biomarkers could fill this void providing the means to decrease therapeutic morbidity, appropriately escalate therapy and serve as measures to benchmark new therapies. PUBLIC HEALTH RELEVANCE: Pulmonary artery hypertension in children or adults (PAH) is a progressive and fatal disease and although there have been advances in therapy and survival with IPAH, the etiology is still largely unknown. In this proposal we will use a state of the art proteomics pipeline to identify and verify specific lung/pulmonary artery endothelial cell prognostic/diagnostic IPAH biomarkers. Therefore biomarkers could fill this void providing the means to decrease therapeutic morbidity, appropriately escalate therapy and serve as measures to benchmark new therapies. (End of Abstract).