Modulation of Macrophage Function through Alternative Splicing in Cardiometabolic Diseases

项目摘要 这个应用程序是为林博士发展她的职业生涯作为一个学术物理学家，科学家专注于 心脏代谢疾病的功能基因组学。职业发展计划包括mRNA培训 处理，巨噬细胞生物学，基因组编辑和人类疾病的动物模型，并提供正式的指导 由Kiran Musunuru博士和丹尼尔雷德博士沿着与一个多学科委员会的杰出 科学家拟议的研究和结构化的指导活动将在佩雷尔曼学校进行， 宾夕法尼亚大学的医学，提供了非常丰富的研究和培训环境 能促进林博士的专业发展拟议的研究将集中在如何关键的替代方案， 剪接（AS）事件有助于心脏代谢疾病的病理生理学， 世界范围内发病率和死亡率都很高。与慢性全身性炎症、心脏代谢性 诸如动脉粥样硬化的疾病涉及巨噬细胞的活化， 促炎表型，但这种转变的机制仍然不完全清楚。 尽管转录水平的变化以前已经被描述过，但我们对转录水平的理解中缺少的一个环节是： 巨噬细胞的可塑性和功能是否涉及转录后调节。的到来 RNA测序（RNA-seq），AS-从单个基因产生多种不同的mRNA亚型-是 作为一种普遍的细胞特异性机制出现，如果失调，可能导致人类疾病。因为 AS在细胞分化中起着重要作用，它可能有助于活化的表型可塑性。 巨噬细胞，并因此心脏代谢疾病的发病机制。尽管很少有研究描述 在巨噬细胞表型的极端亚型水平的差异，我们的初步RNA-seq研究， 活化和静息的人类单核细胞衍生的巨噬细胞已经鉴定了200多种AS事件， 与炎症和细胞存活相关的基因。该提案概述了一项研究AS事件如何在 巨噬细胞的两个特定基因有助于炎症和代谢途径相关， 心脏代谢疾病，特别是动脉粥样硬化。第一个基因是PLD 1，已知它在以下方面发挥作用： 脂质加工，并在促炎性巨噬细胞中经历外显子16的增加的包含。第二 基因是ZC 3 HC 1，其在其选择性外显子8中包含与冠状动脉相关的遗传变异体， 疾病拟议的研究包括两个平行的目标，将调查这些基因中的每一个如何作为 改变（1）巨噬细胞炎性表型，（2）与基因功能相关的代谢途径， 动脉粥样硬化的形成;（3）体内动脉粥样硬化的发生和发展。在承接 提出的研究和培训计划，林博士将阐明巨噬细胞功能的新机制， 心脏代谢背景，成熟的功能基因组学独立翻译研究者， 准备成功竞争R 01基金，用于未来的精准医学项目。

PROJECT SUMMARY This application is for Dr. Jennie Lin to develop her career as an academic physician-scientist focusing on the functional genomics of cardiometabolic diseases. The career development plan includes training in mRNA processing, macrophage biology, genome editing, and animal models of human disease, with formal mentoring by Dr. Kiran Musunuru and Dr. Daniel Rader along with a multi-disciplinary committee of distinguished scientists. The proposed studies and structured mentored activities will take place at the Perelman School of Medicine at the University of Pennsylvania, which offers a remarkably rich research and training environment that will foster Dr. Lin's professional development. The proposed research will focus on how key alternative splicing (AS) events contribute to the pathophysiology of cardiometabolic diseases, which continue to incur significant morbidity and mortality worldwide. Associated with chronic systemic inflammation, cardiometabolic diseases such as atherosclerosis involve the activation of macrophages to shift toward a causally implicated pro-inflammatory phenotype, but the mechanisms underlying this shift remain incompletely understood. Although transcript-level changes have previously been characterized, a missing link in our understanding of macrophage plasticity and function is whether post-transcriptional regulation is involved.! With the advent of RNA sequencing (RNA-seq), AS − the generation of multiple different mRNA isoforms from a single gene − is emerging as a pervasive cell-specific mechanism that can lead to human diseases if dysregulated. Because AS plays an impactful role in cellular differentiation, it may contribute to the phenotypic plasticity of activated macrophages and hence the pathogenesis of cardiometabolic diseases. Although few studies describe isoform-level differences in the extremes of macrophage phenotypes, our preliminary RNA-seq studies of activated and resting human monocyte-derived macrophages have identified more than 200 AS events for genes associated with inflammation and cell survival. This proposal outlines a plan to study how AS events in macrophages for two specific genes contribute to inflammatory and metabolic pathways relevant to cardiometabolic diseases and, in particular, atherosclerosis. The first gene is PLD1, which has known roles in lipid processing and undergoes increased inclusion of exon 16 in pro-inflammatory macrophages. The second gene is ZC3HC1, which houses in its alternative exon 8 a genetic variant associated with coronary artery disease. The proposed studies include two parallel aims that will investigate for each of these genes how AS alters (1) macrophage inflammatory phenotype, (2) metabolic pathways relevant to gene function and atherogenesis, and (3) the development and progression of atherosclerosis in vivo. In undertaking the proposed studies and training plan, the Dr. Lin will elucidate novel mechanisms of macrophage function in the cardiometabolic context, mature as an independent translational investigator in functional genomics, and prepare to compete successfully for R01 funding for future projects aimed at precision medicine.