RNA-Binding Proteins in the Regulation of Vascular Inflammation and Immunity

RNA 结合蛋白在血管炎症和免疫调节中的作用

基本信息

批准号：
10339436
负责人：
Patrick Andries Murphy
金额：
$ 55.82万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-15 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10339436
关键词：
Affect Alternative Splicing Aneurysm Arterial Fatty Streak Arterial Intimas Atherosclerosis Behavior Binding Binding Proteins Biological Biological Assay Blood Vessels CD8-Positive T-Lymphocytes CRISPR screen Cardiovascular Diseases Cardiovascular system Cell Adhesion Molecules Cells Chronic Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques DNA Transposable Elements Data Dependence Disease Endothelial Cells Endothelium Equilibrium Event Evolution Exons Extracellular Matrix Proteins Family Fibronectins Flow Cytometry Future Genes Genetic Transcription Genetic Variation Genome Histology Human Human Genome Immune Immune system Immunity In Vitro Inflammation Inflammatory Inflammatory Response Injury Introns Knowledge Lesion Link Long Interspersed Elements Mediating Modeling Nuclear RNA Pathogenicity Pattern Post-Transcriptional Regulation Process Property Proteomics Quantitative Trait Loci RNA RNA Binding RNA Splicing RNA-Binding Proteins Regulation Regulator Genes Risk Role Rupture Shapes Signal Transduction Site Sterility Stress System T cell differentiation T cell therapy T-Lymphocyte Testing Therapeutic Tissue Preservation Transcript Transgenic Mice Variant Vascular Cell Adhesion Molecule-1 cytokine design extracellular falls genomic locus immune function in vivo in vivo evaluation insight intercellular cell adhesion molecule link protein monocyte mouse genome novel oxidized low density lipoprotein pathogen programs recruit response vascular inflammation

项目摘要

Inflammation is designed to destroy, disable, or contain pathogenic invaders, but must be controlled to avoid destruction of key host systems, like the vasculature. When the interaction between immune cells and the vasculature goes awry, it can contribute to vascular lesions in aneurysm, atherosclerosis, and other diseases. Our study of the interactions between innate immune cells and the arterial wall in models of atherosclerosis – a sterile and chronic injury process with a critical inflammatory component – has revealed broad regulation of alternative splicing responses that change the extracellular composition of the inflamed intima and the behavior of recruited immune cells that protect the arterial wall from damage. Guided by these data and novel in vitro CRISPR screens to probe the function of RNA binding proteins (RBP) in the regulation endothelial inflammation, we have discovered a set of RBP responsive to innate immune cell recruitment that are critical in orchestrating the activation of the endothelium through NFkB signaling. Here, we test the hypothesis that one of these RBP, Elavl1, coordinates alternative splicing in the arterial intima in response to innate immune cell recruitment to regulate chronic immune functions (Aim 1). In seeking a deeper understanding of this immune-regulatory system, we made the unexpected discovery that, like Elavl1, many RBP strongly bind to transposable element (TE) sequences inserted within genes and their RNA transcripts (p<0.0001). While most TE are inactive, these vestigial TE sequences account for ~45% of our genome, are found in nearly all genes, and can provide cryptic splice sites in transcripts that depend on RBP activity. Thus, we aim to define the family of TE-binding RBP, to understand their regulation during inflammatory responses, and their impact on splicing patterns and inflammatory responses through binding to TE (Aim 2). The completion of these aims will provide new insight into the contribution of endothelial alternative splicing responses to inflammation in chronic inflammatory states, and the contribution of pervasive TE-derived sequence to transcript regulation through RBP that bind them, providing new avenues to understand and treat chronic inflammation in the cardiovascular system.

炎症旨在破坏，禁用或包含致病性入侵者，但必须控制避免破坏关键宿主系统，例如脉管系统。当免疫细胞与脉管系统出现问题，它可能导致动脉瘤，动脉粥样硬化和其他的血管病变疾病。我们研究先天免疫细胞与动脉壁之间的相互作用的研究动脉粥样硬化 - 一种无菌和慢性损伤过程，具有关键的炎症成分 - 已显示对改变发炎的细胞外组成的替代剪接反应的广泛调节内膜和保护动脉壁免受损伤的招募的免疫细胞的行为。指导这些数据和新颖的体外CRISPR筛选，以探测RNA结合蛋白（RBP）在调节内皮炎症，我们发现了一组对先天免疫细胞反应的RBP 募集对于通过NFKB信号来策划内皮的激活至关重要。这里，我们检验了以下假设：这些RBP elavl1之一坐在动脉内膜中的替代剪接对先天免疫细胞募集的反应以调节慢性免疫功能（AIM 1）。在寻求一个对这种免疫调节系统的更深入了解，我们提出了一个意外的发现，就像 Elavl1，许多RBP与基因及其中插入的可转座元件（TE）序列强烈结合 RNA转录本（p <0.0001）。虽然大多数TE是不活动的，但这些遗传序列占约45％我们的基因组几乎在所有基因中都发现，并且可以在依赖的转录本中提供加密剪接位点 RBP活动。这是，我们旨在定义Te-Binding RBP的家庭，以了解他们在炎症反应及其对剪接模式和炎症反应的影响到TE（目标2）。这些目标的完成将为内皮的贡献提供新的见解慢性炎症状态下对炎症的替代剪接反应，以及通过将它们绑定的rbp对成绩单调节的普遍衍生的序列，提供了新的途径了解和治疗心血管系统中的慢性炎症。