ADAR mediated RNA editing is a causal mechanism in coronary artery disease

ADAR 介导的 RNA 编辑是冠状动脉疾病的因果机制

• 批准号: 10629687
• 负责人: Chad S Weldy
• 金额: $ 16.6万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629687
• 关键词: ADAR1; Acceleration; Address; Arteries; Atherosclerosis; Bioinformatics; Breeding; CRISPR interference; Cardiovascular system; Cause of Death; Cell Line; Cell Lineage; Cells; Clinical; Clonality; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Coronary Arteriosclerosis; Coronary artery; DNA sequencing; Data; Deficiency Diseases; Development; Disease; Double-Stranded RNA; Enzymes; Epigenetic Process; Evaluation; Foundations; Frequencies; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Diseases; Genetic Transcription; Genetic Variation; Genomic approach; Human; Human Genetics; In Vitro; Inflammation; Inflammatory Bowel Diseases; Interferons; Knowledge; Lipids; Lupus; Maps; Mediating; Medicine; Mentors; Molecular Biology; Mus; Nature; Pathogenesis; Patients; Phenotype; Physicians; Play; Process; Quantitative Trait Loci; RNA; RNA Editing; Role; Scientist; Signal Transduction; Smooth Muscle Myocytes; Testing; Therapeutic; Tissues; Training; Variant; Vascular Diseases; Vascular Smooth Muscle; Vascular calcification; Work; autoinflammatory diseases; biobank; calcification; career; causal variant; design; disorder risk; epigenomic profiling; gene discovery; gene network; genome wide association study; genomic locus; in vivo; inhibitor; innovation; knock-down; loss of function; mouse model; novel; novel therapeutics; programs; rare mendelian disorder; response; sensor; single-cell RNA sequencing; skills; small molecule; therapeutic target; transcriptome sequencing; vascular inflammation; whole genome

PROJECT SUMMARY/ABSTRACT Coronary artery disease (CAD) is the leading cause of death worldwide and there remains a crucial need to discover mechanisms of disease and develop new therapies. Leveraging the power of human genetics and mechanistic discovery, we are poised to understand causal mechanisms of disease and design critical therapies. Although there are more than 160 genetic loci discovered which are associated with CAD, outside of lipid lowering therapies, the promise of these genome wide association studies (GWAS) to identify causal genes and result in novel mechanisms for treatment has not been fulfilled. Recent discoveries have revealed that RNA editing is a fundamental mechanism of inflammation and disease, a process mediated by ADAR enzymes. In a groundbreaking discovery now accepted in Nature, the lab of the applicant’s co-mentor, Dr. Jin Billy Li, has discovered that common genetic variation which decreases RNA editing powerfully increases CAD risk. Preliminary data implicates RNA editing in vascular smooth muscle cells (SMCs) to have a causal role in CAD. In this proposal, the applicant aims to elucidate the fundamental mechanisms of ADAR mediated RNA editing, and the potential for targeting the pivotal downstream double stranded RNA (dsRNA) sensor, MDA5 (encoded by IFIH1), as treatment in CAD. Vascular inflammation has long been implicated in the onset and progression of atherosclerosis, but the specific disease modifying targets remain elusive. Pivotal studies have discovered that endogenous dsRNA is formed in normal RNA transcription and undergoes RNA editing by ADAR1. If insufficiently edited, dsRNA will activate a powerful interferon stimulated gene (ISG) response through MDA5 (IFIH1). Common genetic variation can modify RNA editing frequency — ‘editing-QTLs’ (edQTL) — where edQTLs are not only predictive of CAD risk, but also other autoinflammatory disorders including lupus and inflammatory bowel disorders. In this proposal, the applicant hypothesizes that the ADAR-MDA5 axis plays an important role in the onset of CAD and that MDA5 is an ideal therapeutic target. However, there is a critical gap in knowledge, and it is unknown if deficient RNA editing in SMCs accelerates the development of atherosclerosis, and it is imperative that the ADAR-MDA5 axis is investigated to reveal its potential for therapeutic innovation. To address this, the applicant has bred a novel SMC lineage traced atherosclerosis mouse model with conditional SMC deletion of Adar with and without Ifih1 deletion. Through a one-of-a-kind biobank of primary HCASMCs, the applicant will further characterize the effect of MDA5 activation from decreased RNA editing. Further, the applicant will discover the key genes downstream of MDA5 activation through a cutting-edge genomics approach using epigenetic editing and single cell RNA sequencing (perturb-seq). This work will advance the applicant’s training and enable the applicant to gain skill sets in bioinformatics, genetics, and molecular biology — launching the applicant’s career as an independent physician-scientist.

项目摘要/摘要 冠状动脉疾病(CAD)是世界范围内的主要死亡原因，仍然迫切需要 发现疾病的机制并开发新的治疗方法。利用人类遗传学的力量 机械发现，我们准备理解疾病的因果机制，并设计关键的 治疗。虽然已经发现了160多个与冠心病相关的遗传基因座，但除了 降脂疗法，这些全基因组关联研究(GWAS)确定病因的前景 基因和结果的新的治疗机制尚未完成。 最近的发现表明，RNA编辑是炎症的基本机制 和疾病，这是一个由ADAR酶介导的过程。这一突破性的发现现已被《自然》杂志接受， 申请人的共同导师金比利·李博士的实验室发现，常见的基因变异 减少RNA编辑会极大地增加CAD风险。初步数据表明血管中存在RNA编辑 平滑肌细胞(SMC)在CAD中起因果作用。在这份提案中，申请人的目标是 阐明ADAR介导的RNA编辑的基本机制，以及靶向 关键下游双链RNA(DsRNA)传感器，MDA5(由IFIH1编码)，用于CAD的治疗。 长期以来，血管炎症一直与动脉粥样硬化的发生和发展有关，但 具体的疾病改善目标仍然难以捉摸。关键研究发现，内源性dsRNA是 在正常的RNA转录中形成，并由ADAR1进行RNA编辑。如果编辑不足，dsRNA将 通过MDA5(IFIH1)激活强大的干扰素刺激基因(ISG)反应。共同遗传 变异可以改变RNA编辑频率--编辑-QTL(EdQTL)--其中edQTL不仅是预测性的 不仅包括冠心病风险，还包括其他自身炎症性疾病，包括狼疮和炎症性肠病。 在这项建议中，申请人假设ADAR-MDA5轴在发病中起重要作用 MDA5是一个理想的治疗靶点。然而，在知识方面存在着一个关键的鸿沟，它是 尚不清楚SMC中的RNA编辑缺陷是否会加速动脉粥样硬化的发展，这是势在必行的 研究ADAR-MDA5轴以揭示其治疗创新的潜力。 为了解决这个问题，申请人培育了一种新的SMC谱系追踪动脉粥样硬化的小鼠模型 在删除和不删除IFIH1的情况下有条件地删除ADAR的SMC。通过一个独一无二的初级生物库 对于HCASMCs，申请人将进一步表征减少RNA编辑而激活MDA5的影响。 此外，申请者将通过尖端技术发现MDA5激活下游的关键基因 使用表观遗传编辑和单细胞rna测序的基因组学方法。这项工作将 推进申请者的培训，使申请者能够获得生物信息学、遗传学和 分子生物学--启动申请者作为独立内科医生-科学家的职业生涯。