Phosphopeptide mapping of plexiform lesions in pulmonary arterial hypertension

肺动脉高压丛状病变的磷酸肽图谱

• 批准号: 8335485
• 负责人: LAWRENCE S. ZISMAN
• 金额: $ 7.25万
• 依托单位: PULMOKINE, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8335485
• 关键词: Address; Algorithms; Bioinformatics; Blood Vessels; Blood flow; Cells; Chromatography; Clinical Trials; Data; Disease; Family; Freezing; Frozen Sections; Goals; Growth; Imatinib; Immunoassay; Incubated; Intervention; Knowledge; Label; Lead; Lesion; Lung; Maps; Mass Spectrum Analysis; Methods; Morbidity - disease rate; Output; Pathology; Patients; Peptides; Phase; Phosphopeptides; Phosphoproteins; Phosphorylation; Phosphorylation Site; Phosphotransferases; Proliferating; Protein Analysis; Protein Isoforms; Protein-Serine-Threonine Kinases; Proteins; Proteomics; Protocols documentation; Public Health; Rare Diseases; Receptor Protein-Tyrosine Kinases; Regulation; Relative (related person); Research; Role; Sampling; Societies; Structure of parenchyma of lung; Techniques; Testing; Tissues; Tyrosine; Tyrosine Kinase Inhibitor; arteriole; blood lead; candidate identification; cell growth; experience; hypertension control; innovation; interest; kinase inhibitor; laser capture microdissection; mortality; nanofluidic; new therapeutic target; novel; oncology; protein aminoacid sequence; pulmonary arterial hypertension; therapeutic target; titanium dioxide; validation studies

Pulmonary Arterial Hypertension (PAH) is a rare disorder of the pulmonary vasculature associated with high morbidity and mortality. The pathology of the disease consists of plexiform lesions of proliferating cells which obstruct blood flow through the pulmonary arterioles. There is a growing interest in the use of kinase inhibitors to address this underlying pathology. However, very little is actually understood about the regulation of kinases in PAH, and the phosphoproteome in PAH has not yet been studied. This project will determine the phosphopeptide profile of lung tissue from subjects with idiopathic pulmonary arterial hypertension (iPAH) compared to controls. Titanium dioxide chromatography will be used to enrich for phosphopeptides which will then be identified by mass spectroscopy. A bioinformatics approach will be used to predict the kinases and/or kinase families most likely to be responsible for these increased phosphoproteins. The NetworKIN and other algorithms will be used for this purpose. The predictions of NetworkKIN will be tested in a subset of kinases by incubating the selected recombinantly expressed kinases with peptide microarrays that duplicate the peptide sequences of the candidate phosphorylation sites. The findings of the proteomic analysis will be validated by isolating protein from plexiform lesions with laser capture microdissection. The proteins will be analyzed with a novel nanofluidic immunoassay that can detect very small quantities of protein. The results of this study could identify new therapeutic targets for the treatment of iPAH and thereby benefit patients and society.

肺动脉高压是一种罕见的肺血管疾病 与高发病率和死亡率相关。该疾病的病理包括丛状 增殖细胞病变，阻塞肺小动脉血流。有 对使用激酶抑制剂来解决这种潜在的病理学的兴趣越来越大。然而，在这方面， 关于PAH中激酶的调节实际上知之甚少， PAH中的磷酸化蛋白质组尚未研究。该项目将决定 特发性肺动脉高压患者肺组织磷酸肽谱 高血压（iPAH）与对照组相比。二氧化钛色谱法将用于 富集磷酸肽，然后通过质谱鉴定。一 生物信息学方法将用于预测激酶和/或激酶家族最可能 导致磷蛋白的增加。NetworkIN和其他算法将 用于此目的。NetworkKIN的预测将在激酶的一个子集中进行测试， 将所选择的重组表达的激酶与肽微阵列一起孵育， 复制候选磷酸化位点的肽序列。的调查结果 蛋白质组学分析将通过激光捕获从丛状病变中分离蛋白质来验证 显微解剖这些蛋白质将用一种新的纳米流体免疫测定法进行分析， 检测到非常少量的蛋白质。这项研究的结果可以确定新的治疗方法， 治疗iPAH的目标，从而造福患者和社会。