Genomics of Pulmonary Vascular Disease

项目总结 肺动脉高压(PH)是一种以进行性肺血管为特征的潜在致命性疾病 重塑和肺动脉压力升高，导致右心衰竭。分子病因学是 即使在骨形态发生蛋白(BMP)信号突变的家族性PH中也知之甚少 途径得到了很好的描述。我们的方法侧重于了解生殖系遗传变异 易患肺高压，肺内的躯体改变可能有助于肺泡炎的发生和/或进展 这种疾病。我们是第一个在人的肺动脉内皮细胞中发现染色体异常的人 取外植体PH肺组织。患者肺和血细胞的DNA损伤水平高于对照组， 并与活性氧含量呈正相关。在他的血液中也发现了类似的变化 患者的亲属，这表明这可能是遗传决定的。我们目前的研究利用了最先进的技术 在肺组织和细胞中进行下一代测序方法以检验以下假设： DNA损伤易导致肺内遗传改变，并可能导致肺血管重塑。 我们还在测试新的治疗方法，以纠正家族性PH和遗传性高血压患者的BMP信号 出血性毛细血管扩张症(HHT)是一种相关的血管疾病，也是由BMP途径的突变引起的。 HHT的发病率很高，与肺动静脉畸形出血的风险有关， 肝脏和大脑。我们目前的研究重点是阿托鲁仑，这是一种具有孤儿药物状态的小分子，它可以促进 无义突变的核糖体通读。在初步研究中，BMP信号在细胞中恢复 6例患者中有5例存在不同的无义突变。血液或肺来源内皮细胞的体外研究 在遗传小鼠模型中，对受影响患者的治疗和预防研究进行了补充。 这一杰出调查者奖提案将这两个主要主题结合到一个统一的研究中 肺血管疾病基因组学计划。我们将利用单细胞的新兴力量 测序技术用于开发粘连在Swan-Ganz导管上的内皮细胞的新分析 常规心导管术后。这将在几个新的方向上扩展我们目前的研究：(1)使 直接分析疾病相关细胞中的DNA损伤标记；(2)提供即时的体外读数 药物反应；(3)发展新的诊断和预后标志；以及(4)可能最终 提供单细胞分辨率的时间和进化的体细胞突变在肺。穿过 在两个由NHLBI资助的国家财团的基因组学组成部分中处于领先地位，我们将利用 这些创新研究并将它们与广泛的其他组学数据相结合，以实现以下目标 提高肺血管疾病的诊断、精准治疗和终极预防水平。

PROJECT SUMMARY Pulmonary hypertension (PH) is a potentially fatal disease characterized by progressive pulmonary vascular remodeling and elevated pulmonary artery pressure, leading to right heart failure. The molecular etiology is poorly understood, even in familial PH where mutations in the bone morphogenetic protein (BMP) signaling pathway are well characterized. Our approach focuses on understanding the germline genetic variation that predisposes to PH, and somatic changes within the lung that may contribute to the onset and/or progression of the disease. We were the first to identify chromosome abnormalities in pulmonary artery endothelial cells from explant PH lung tissue. Levels of DNA damage were higher in lung and blood cells from patients than controls, and correlated with the amount of reactive oxygen species. Similar changes were found in the blood of patients' relatives, suggesting this may be genetically determined. Our current studies utilize state of the art next generation sequencing approaches in lung tissues and cells to test the hypothesis that increased levels of DNA damage predispose to genetic alterations in the lung, and may contribute to vascular remodeling in PH. We are also testing novel therapeutic approaches to correct BMP signaling in familial PH and hereditary hemorrhagic telangiectasia (HHT), a related vascular disorder also caused by mutations in the BMP pathway. HHT carries high morbidity associated with risk of hemorrhage from arteriovenous malformations in the lungs, liver and brain. Our current studies focus on ataluren, a small molecule with orphan drug status that promotes ribosomal readthrough of nonsense mutations. In preliminary studies, BMP signaling was restored in cells from 5 of 6 patients with different nonsense mutations. In vitro studies in blood or lung-derived endothelial cells from affected patients are complimented with treatment and prevention studies in a genetic mouse model. This Outstanding Investigator Award proposal combines these two major themes into a unified research program in the genomics of pulmonary vascular disease. We will harness the emerging power of single cell sequencing technologies to develop novel analyses of endothelial cells that adhere to the Swan-Ganz catheter after routine cardiac catheterization. This will extend our current studies in several new directions: (1) enabling direct analysis of DNA damage markers in disease-relevant cells; (2) providing immediate ex vivo readouts of drug responses; (3) developing novel diagnostic and prognostic signatures; and (4) perhaps ultimately providing single cell resolution of the timing and evolution of somatic mutations in the PH lung. Through leadership in the genomics components of two NHLBI-funded national consortia, we will leverage the results of these innovative studies and integrate them with a broad range of other omics data to realize the goals of improving the diagnosis, precision treatment and ultimate prevention of pulmonary vascular disease.