Integrative Multiomics to Uncover Novel Genes and Networks in Pulmonary Arterial Hypertension

项目摘要/摘要 这项建议描述了一个有指导的医生-科学家培训计划，以发现新的基因和网络 在肺动脉高压(PAH)中使用整合的多组学方法。候选人目前 在大卫·格芬医学院肺和重症监护科发展他的学术生涯 加州大学洛杉矶分校的医学。他的长期目标是为知情的PAH患者开发更有效的靶向治疗 通过对致病机制的更深入的了解。在他的导师曼苏雷博士的指导下 Eghbali和夏阳，候选人将在集成系统方面发展独特的跨学科技能， 单细胞、空间和实验生物学，将促进他在该领域向研究独立性的过渡 多环芳烃。 PAH仍然是一种不治之症，其特征是不可逆的肺血管重构，质量差 生命，守护着长期的预后。利用强大的团队整合最新的经济学和 迫切需要计算方法来确定PAH肺中潜在的候选分子驱动因素 治疗靶点。获得人类PAH最大生物库的RNA测序，并对照肺 Date(n=148)，我们已经通过共表达网络分析确定了由266个基因组成的模块(我们指的是 如“粉红色”模块)，这与PAH肺密切相关。通过右心多模式整合 导尿术数据、组织学分析和全基因组关联研究(GWAS)，我们发现粉红色 模块不仅在PAH肺中转录上调，而且与血流动力学增强有关 PAH的严重性、血管重塑和遗传风险。我们的初步数据显示，粉色模块基因是1) 肺血管细胞的调节失调，2)与肺血管重塑相关的途径丰富 如内皮细胞-间充质转化和Wnt信号，3)和可能是 PAH，如ANTXR1，一种整合素样糖蛋白，与多种癌症密切相关，但从未被研究过 啊哈。鉴于越来越多的初步证据表明大量肺源性粉色模块的重要性，更深层次的 需要研究其在PAH发病中的细胞特异性作用，以促进我们对PAH的理解 PAH肺的分子驱动因素，并确定新的治疗靶点。我们假设粉色模块驱动 PAH肺血管细胞内调节失调导致的血管重构。为了检验这一假设， 我们将1)解决特定的细胞环境中，其中粉红色模块是失调的PAH肺使用单一- 核RNAseq和空间转录和2)确定体外敲除粉色模块的效果 PAH肺血管细胞中的候选驱动基因，如ANTXR1。拟议的研究将利用 结合尖端的多组学方法和实验生物学，更深入地了解 来自大型肺生物库的新的PAH相关基因集，将为我自己的实验室提供基础 以及未来的R01，它将专注于基础机械和翻译研究。

Project Summary/Abstract This proposal describes a mentored physician-scientist training program to uncover novel genes and networks in pulmonary arterial hypertension (PAH) using an integrative multiomics approach. The candidate is currently developing his academic career in the Division of Pulmonary & Critical Care at the David Geffen School of Medicine of UCLA. His long-term goal is to develop more effective targeted therapies for PAH patients informed by a deeper knowledge of the pathogenic mechanisms. Under the guidance of his mentors Drs. Mansoureh Eghbali and Xia Yang, the candidate will develop a unique cross-disciplinary skillset in integrative systems, single-cell, spatial and experimental biology that will facilitate his transition to research independence in the field of PAH. PAH remains an incurable disease characterized by irreversible pulmonary vascular remodeling, poor quality of life, and guarded long-term prognosis. Leveraging a well-powered cohort integrating the latest omics and computational methodologies is critically needed to identify candidate molecular drivers in PAH lungs as potential therapeutic targets. With access to RNA sequencing of the largest biobank of human PAH and control lungs to date (n=148), we have identified, by co-expression network analysis, a module of 266 genes (which we refer to as the “pink” module) that is strongly associated with PAH lungs. Through multimodal integration with right heart catheterization data, histological analyses, and genome-wide association studies (GWAS), we found the pink module is not only transcriptionally upregulated in PAH lungs, but also associated with increased hemodynamic severity, vascular remodeling, and genetic risk of PAH. Our preliminary data suggests pink module genes are 1) dysregulated in pulmonary vascular cells, 2) enriched in pathways relevant to pulmonary vascular remodeling such as endothelial-mesenchymal transition and Wnt signaling, 3) and may be candidate molecular drivers of PAH, such as ANTXR1, an integrin-like glycoprotein strongly implicated in various cancers but never studied in PAH. Given the mounting preliminary evidence for the importance of the bulk lung-derived pink module, a deeper investigation into its cell-specific role in PAH pathogenesis is needed to advance our understanding of the molecular drivers of PAH lungs and identify new therapeutic targets. We hypothesize that the pink module drives vascular remodeling in PAH through its dysregulation within pulmonary vascular cells. To test this hypothesis, we will 1) resolve the specific cellular context in which the pink module is dysregulated in PAH lungs using single- nucleus RNAseq and spatial transcriptomics and 2) determine the effects of in vitro knockdown of a pink module candidate driver gene, such as ANTXR1, in PAH pulmonary vascular cells. The proposed studies will utilize a combination of cutting-edge multiomic approaches and experimental biology to provide greater insight into a novel PAH-associated gene set derived from a large lung biobank, and will provide a foundation for my own lab and future R01 that will focus on basic mechanistic and translational studies.