Isolation/Characterization CECs Pulmonary Arteriopathy

肺动脉病 CEC 的分离/表征

• 批准号: 7003995
• 负责人: michael a solomon
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7003995
• 关键词: RNA; biomarker; cell surface receptors; diagnosis design /evaluation; early diagnosis; flow cytometry; heart catheterization; human subject; microarray technology; monocyte; nucleic acid amplification techniques; oligonucleotides; patient oriented research; pulmonary hypertension; vascular endothelium

Primary pulmonary hypertension (PPH), a subgroup of plexogenic pulmonary arteriopathy (PPA), is a rare disorder associated with severe morbidity and high mortality rates. There are no routine screening tests or validated markers of disease activity in PPH, or the broader group of PPA. Therefore, patients usually present at advanced stages of disease. The pathogenesis of PPH and other forms of PPA remain unclear. Current thinking focuses on a "two-hit" hypothesis: 1) genetic susceptibility, and 2) a triggering stimulus that initiates pulmonary vascular injury, resulting in endothelial cell dysfunction. Endothelial cells are normally shed into the circulation and are a valuable source of clinical material for studying diseases characterized by endothelial cell dysfunction. Unfortunately, no clear methodology exists for isolating clinically relevant numbers of circulating endothelial cells (CECs). In the bench phase of the project we plan to use flow cytometry to develop a methodology for isolating clinically relevant numbers of viable CECs. We hypothesize that CECs can be used to define a subset of differentially regulated biomarkers in PPH and other forms of PPA that may lead to earlier diagnosis and better methods for measuring responses to therapy. We also hope to identify novel targets for future therapeutic interventions. In the clinical phase of the project, we will recruit the following subject groups: 1) patients with newly diagnosed PPH and other forms of PPA, 2) patients with pulmonary hypertension (PH) ascribed to a nonvascular injury process and 3) normal individuals (controls). All subjects will undergo right heart catheterization. CECs drawn peripherally and from the pulmonary artery catheter will be characterized for disease phenotype by cell surface markers and oligonucleotide microarrays. Total RNA for microarrays will be prepared from CECs by cell sorting and subjected to amplification. In addition endothelial progenitor cells will be quantitated and peripheral blood mononuclear cells (PBMCs) will be isolated. PBMCs will be studied in depth using high density oligonucleotide microarrays to more fully characterize their transcriptome. We plan to follow response to therapy by restudying the same parameters in patients with PPH or PPA after therapeutic intervention.

原发性肺动脉高压（PPH）是丛源性肺动脉病（PPA）的一个亚类，是一种罕见的疾病，具有严重的发病率和高死亡率。 PPH或更广泛的PPA组中没有常规的筛查试验或有效的疾病活动标志物。 因此，患者通常处于疾病的晚期阶段。PPH和其他形式的PPA的发病机制尚不清楚。目前的想法集中在“两击”假说：1）遗传易感性，和2）触发刺激，启动肺血管损伤，导致内皮细胞功能障碍。 内皮细胞通常脱落到循环中，并且是用于研究以内皮细胞功能障碍为特征的疾病的有价值的临床材料来源。 不幸的是，没有明确的方法存在用于分离临床相关数量的循环内皮细胞（CEC）。在该项目的实验室阶段，我们计划使用流式细胞术开发一种方法来分离临床相关数量的活CEC。 我们假设CEC可用于定义PPH和其他形式的PPA中差异调节的生物标志物的子集，这可能导致早期诊断和更好的方法来测量对治疗的反应。 我们还希望为未来的治疗干预确定新的靶点。 在该项目的临床阶段，我们将招募以下受试者组：1）新诊断的PPH和其他形式的PPA患者，2）归因于非血管损伤过程的肺动脉高压（PH）患者和3）正常个体（对照）。 所有受试者将接受右心导管插入术。将通过细胞表面标志物和寡核苷酸微阵列表征外周和从肺动脉导管抽取的CEC的疾病表型。 将通过细胞分选从CEC制备用于微阵列的总RNA并进行扩增。 此外，将对内皮祖细胞进行定量，并分离外周血单核细胞（PBMC）。将使用高密度寡核苷酸微阵列深入研究PBMC，以更全面地表征其转录组。我们计划通过重新研究PPH或PPA患者在治疗干预后的相同参数来跟踪治疗反应。