Chronic Thromboembolic Pulmonary Hypertension

慢性血栓栓塞性肺动脉高压

• 批准号: 7037448
• 负责人: TIMOTHY A MORRIS
• 金额: $ 15.07万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7037448
• 关键词: clinical research; computer assisted sequence analysis; crosslink; fibrinogen; fibrinolysis; gene expression; human subject; isozymes; liquid chromatography mass spectrometry; nucleic acid sequence; pathologic process; patient oriented research; plasmin; polymerase chain reaction; posttranslational modifications; protein sequence; protein structure function; pulmonary hypertension; thromboembolism; thrombosis

DESCRIPTION (provided by applicant): Chronic thrombo-embolic pulmonary hypertension (CTEPH) is characterized by progressive right ventricular failure and, if left untreated, death. It is preceded by acute pulmonary thrombo-embolism. Unlike other patients with acute pulmonary embolism, however, patients with CTEPH do not lyse the thrombotic material within the pulmonary arteries. Instead, the thrombi are remodeled into intravascular scars that obstruct flow, often with disastrous consequences. Our goal is to understand the mechanisms responsible for the failure to resolve the acute thrombo-emboli in CTEPH. Our previous studies had disclosed no defect within fibrinolytic enzymes in these patients. However, we demonstrated that fibrinogen purified from CTEPH patients is somewhat resistant to lysis by exogenous plasmin. For this reason, we plan to search within the CTEPH patient population for genetic variants of fibrinogen and post-translational modifications which might make it resistant to fibrinolysis. Fibrinogen variants will be searched for in CTEPH patients using three complementary methods. First, purified fibrinogen will be polymerized and cross-linked to simulate normal thrombosis, then exposed to plasmin under controlled conditions to determine the profile of peptide fragments yielded during fibrinolysis, as well as the kinetics of their release. Next, genetic variants in targeted portions of fibrinogen will be looked for using the polymerase chain reaction (PCR) and DMA sequencing. Finally, primary amino acid sequence and post translational modifications will be searched for in fibrinogen from CTEPH, using purified fibrinogen samples fragmented with proteases. The fragments will be subjected to liquid chromatography-mass spectroscopy with data-dependent MS-2 data acquisition, followed by computer-assisted sequence analysis.

描述（由申请人提供）：慢性血栓栓塞性肺动脉高压（CTEPH）的特征是进行性右心室衰竭，如果未治疗，则为死亡。在此之前是急性肺血栓栓塞。但是，与其他急性肺栓塞患者不同，CTEPH患者不会裂解肺动脉内的血栓形成材料。取而代之的是，将血栓重塑为阻碍流动的血管内疤痕，通常会带来灾难性的后果。我们的目标是了解导致无法解决CTEPH中急性血栓栓塞的机制。我们以前的研究在这些患者的纤维蛋白水解酶内没有透露缺陷。但是，我们证明了从CTEPH患者纯化的纤维蛋白原对外源性纤溶酶的裂解具有抗性。因此，我们计划在CTEPH患者人群中搜索纤维蛋白原和翻译后修饰的遗传变异，这可能使其对纤维蛋白溶解具有抗性。将使用三种互补方法在CTEPH患者中搜索纤维蛋白原变异。首先，将纯化的纤维蛋白原聚合并交联以模拟正常的血栓形成，然后在受控条件下暴露于纤溶酶，以确定纤维蛋白溶解过程中产生的肽片段的特征以及释放的动力学。接下来，将寻找纤维蛋白原靶向部分中的遗传变异，以使用聚合酶链反应（PCR）和DMA测序。最后，使用纯化的纤维蛋白原样品碎片，将纤维蛋白原样品从CTEPH中搜索初级氨基酸序列和翻译后修饰。这些片段将通过数据依赖的MS-2数据采集进行液相色谱 - 质量光谱，然后进行计算机辅助序列分析。

项目成果

Characterizing pulmonary blood flow distribution measured using arterial spin labeling.

• DOI: 10.1002/nbm.1407
• 发表时间: 2009-12
• 期刊: NMR IN BIOMEDICINE
• 影响因子: 2.9
• 作者: Henderson, A. Cortney;Prisk, G. Kim;Levin, David L.;Hopkins, Susan R.;Buxton, Richard B.
• 通讯作者: Buxton, Richard B.