Risk stratification in pulmonary arterial hypertension: Intersection of OMICs and longitudinal phenotypes through the PAH Biobank

肺动脉高压(PAH)是一种罕见的致命疾病，其特征是逐渐闭塞。 肺小动脉导致肺血管阻力进行性增加并伴有右心恶化 失败和死亡。虽然在少数患者中报告了罕见的突变(例如，BMPR2)，但大多数 患者没有携带已确定的突变。我们最近在2085年发表了关于常见基因变异的文章 使用全基因组关联的特发性/遗传性(I/H)PAH病例和9,659名欧洲血统对照 (GWA)方法。发现和复制分析在四个独立的队列中进行， 我们位于美国的PAH生物库(PAHB)研究和三个国际队列的基因分型阵列 基因组序列数据。我们报道了两个新的基因座，分别位于HLADPA1/DPB1和SOX17附近，与I/H PAH相关。 HL A-DPA/DPB1基因座预测I/H PAH的年死亡率降低25-37%。铅SOX17变种 位于内皮细胞(EC)中与SOX17基因空间位置相近的一个假定的增强子区域 前体，这影响了它的表达基于我们的实验验证。我们的发现提供了第一个 支持共同遗传变异对多环芳烃风险的贡献，并结合最近报告的 关于PAH中SOX17罕见突变的数据，突出了SOX17在PAH中的因果作用。超越PAH风险，我们现在 假设多环芳烃的进展和结果也是转基因的，包括来自SOX17，a 转录因子和人类白细胞抗原-DPA1/DPB1作为新的候选基因和可能的治疗靶点。至 验证这一假设，我们已经开发了3个特定目标(SA)，这将进一步扩大PAHB，世界上 最大的PAH生物库、注册和具有完整外显子组测序(WES)的多组学数据集，RNAseq，Whole 基因组基因分型，以及几乎所有受试者的非靶向代谢组学数据。SA#1将收集序列号 PAHB中的纵向数据，以询问疾病风险基因座(SOX17，HLA-DPA/B1)与 PAH进展的标志物。我们还将评估表达(EQTL)和代谢组学(MQTL)数量性状 用于功能验证的基因座分析。除了疾病风险SOX17/人类白细胞抗原基因座，我们还产生了额外的初步 数据揭示了7个新的与帕金森病患者生存直接相关的遗传基因座的全基因组意义 第二个独立的PAH队列。SA#2现在将复制这些新发现和结果(进展和 在PAHB中收集的SA#1。作为这项建议的一个独特特征，我们将在两个 其他全球PAH队列与可用的eQTL和mQTL数据，并对所有队列进行荟萃分析。我们会 同时构建了一个结合临床危险因素和遗传学(SOX17、HLA7个SNPs)的PAH风险分层工具 结果。最后，基于SOX17在EC功能中的保护作用的初步数据，SA#3将验证 SOX17信号通路在PAH患者ECs/SMC中的生物学作用 以及在小鼠PH模型上的临床前测试。这两个新基因座具有很强的临床相关性 对临床结果预测、临床试验设计和新药靶点开发的影响。

Pulmonary arterial hypertension (PAH) is a rare, fatal condition characterized by the gradual occlusion of the pulmonary arterioles leading to progressively increased pulmonary vascular resistance with worsening right heart failure and death. While rare mutations (e.g., in BMPR2) have been reported in a minority of patients, most patients carry no established mutations. We recently published on common genetic variation in 2,085 idiopathic/heritable (I/H) PAH cases and 9,659 controls of European ancestry using a genome-wide association (GWA) approach. Discovery and replication analyses were conducted in four independent cohorts with genotyping arrays from our US-based PAH Biobank (PAHB) study and three international cohorts with whole genome sequence data. We reported two novel loci, at HLA-DPA1/DPB1 and near SOX17 associated I/H PAH. HLA-DPA/DPB1 locus predicts a reduced annual mortality rate by 25-37% in I/H PAH. The lead SOX17 variant is located in a putative enhancer region in close spatial proximity to the SOX17 gene in endothelial cell (EC) precursors, which influences its expression based on our experimental validation. Our findings provide the first support for the contribution of common genetic variance to PAH risk and, combined with the recently reported data on rare mutations in SOX17 in PAH, highlight the causal role of SOX17 in PAH. Beyond PAH risk, we now hypothesize that PAH progression and outcomes are also genetically modified including from SOX17, a transcription factor, and HLA-DPA1/DPB1 as both novel candidate genes and possible therapeutic targets. To test this hypothesis, we have developed 3 specific aims (SAs) that will further expand the PAHB, the world’s largest PAH biobank, registry, and multi-omics dataset with whole exome sequencing (WES), RNAseq, whole genome genotyping, and non-targeted metabolomics data on nearly all subjects. SA #1 will collect serial longitudinal data in PAHB to interrogate associations between disease risk loci (SOX17, HLA-DPA/B1) with markers of PAH progression. We will also evaluate expression (eQTL) and metabolomics (mQTL) quantitative trait loci analyses for functional validation. Beyond disease risk SOX17/HLA loci, we generated additional preliminary data revealing genome-wide significance for seven novel genetic loci associated directly with survival in PAH in a second, independent PAH cohort. SA #2 will now replicate these new findings with outcomes (progression and survival) collected in PAHB from SA #1. As a unique feature of this proposal, we will interrogate our top loci in two other global PAH cohorts with available eQTL and mQTL data and perform a meta-analysis of all cohorts. We will also construct a risk stratification tool combining clinical risk factors and genetics (SOX17, HLA, 7 SNPs) for PAH outcomes. Finally, based on preliminary data on the protective role of SOX17 in EC function, SA #3 will validate the biological role of SOX17 pathway in the development of PAH using ECs/SMCs isolated from PAH patients as well as pre-clinical testing in murine PH models. Strong clinical association of these two new loci has implications for prediction of clinical outcomes, clinical trial design, and the development of novel drug targets.