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.

炎症旨在摧毁、禁用或遏制病原体入侵者，但必须加以控制当免疫细胞与血管系统相互作用时，避免破坏关键的宿主系统。血管系统出现问题，可能导致动脉瘤、动脉粥样硬化和其他疾病的血管病变我们在疾病模型中研究先天免疫细胞与动脉壁之间的相互作用。动脉粥样硬化——一种具有关键炎症成分的无菌慢性损伤过程——已经揭示广泛调节可变剪接反应，改变发炎细胞的细胞外组成内膜和招募的免疫细胞的行为，保护动脉壁免受损伤。这些数据和新颖的体外 CRISPR 筛选可探测 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有反应的RBP 招募对于通过 NFkB 信号传导协调内皮细胞的激活至关重要。我们测试了这样的假设：这些 RBP 之一 Elavl1 协调动脉内膜中的选择性剪接对先天免疫细胞募集的反应以调节慢性免疫功能（目标 1）。随着对这种免疫调节系统的深入了解，我们有了意想不到的发现，比如 Elavl1，许多 RBP 与插入基因及其内的转座元件 (TE) 序列强烈结合 RNA 转录本 (p<0.0001) 虽然大多数 TE 不活跃，但这些残留的 TE 序列约占 45%。我们的基因组，几乎存在于所有基因中，并且可以在转录本中提供隐秘的剪接位点，这些位点依赖于因此，我们的目标是定义 TE 结合 RBP 家族，以了解它们在过程中的调节。炎症反应及其通过结合对剪接模式和炎症反应的影响 TE（目标 2）的完成将为内皮细胞的贡献提供新的见解。慢性炎症状态下炎症的选择性剪接反应以及普遍的 TE 衍生序列通过结合它们的 RBP 进行转录调控，提供了新的途径了解和治疗心血管系统的慢性炎症。