Genomics of Pulmonary Vascular Disease

项目摘要 肺动脉高压（PH）是一种以进行性肺血管扩张为特征的潜在致死性疾病 重塑和肺动脉压力升高，导致右心衰竭。分子病因学是 即使在家族性PH中，骨形态发生蛋白（BMP）信号转导中的突变也知之甚少， 路径被很好地表征。我们的方法侧重于了解生殖系遗传变异， 易患PH，肺内的躯体变化可能导致PH的发作和/或进展。 这种疾病我们是第一个发现肺动脉内皮细胞染色体异常的人， 取出PH肺组织。患者肺和血细胞中的DNA损伤水平高于对照组， 并与活性氧的量相关。在血液中发现了类似的变化 患者的亲属，这表明这可能是由基因决定的。我们目前的研究利用了最先进的 下一代测序方法在肺组织和细胞中的应用， DNA损伤易导致肺的遗传改变，并可能导致PH的血管重塑。 我们还在测试新的治疗方法，以纠正家族性PH和遗传性PH中的BMP信号传导。 出血性毛细血管扩张症（HHT），一种也由BMP途径中的突变引起的相关血管病症。 HHT具有与肺动静脉畸形出血风险相关的高发病率， 肝脏和大脑我们目前的研究重点是阿他卢伦，这是一种具有孤儿药地位的小分子，可以促进 无义突变的核糖体通读。在初步的研究中，BMP信号在细胞中恢复， 5/6例患者有不同的无义突变。血液或肺源性内皮细胞的体外研究 从受影响的患者补充治疗和预防研究的遗传小鼠模型。 本次杰出研究者奖提案将这两大主题结合为统一的研究 肺血管疾病基因组学研究项目。我们将利用单细胞的新兴力量 测序技术开发粘附于Swan-Ganz导管的内皮细胞的新分析 在常规心导管插入术之后。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损伤标志物;（2）提供以下信息的即时离体读数： 药物反应;（3）开发新的诊断和预后标志;（4）可能最终 提供PH肺中体细胞突变的时间和演变的单细胞分辨率。通过 领导两个NHLBI资助的国家财团的基因组学组成部分，我们将利用的结果， 这些创新的研究，并将其与广泛的其他组学数据，以实现目标， 提高肺血管疾病的诊断、精确治疗和最终预防。

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.