Definition of Serum Ribonucleoprotein Composition and its Regulation and Function

血清核糖核蛋白组成的定义及其调控和功能

• 批准号: 8912881
• 负责人: THOMAS TUSCHL
• 金额: $ 140.88万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8912881
• 关键词: Affect; Age; Amyloid; Antibodies; Archives; Autoantigens; Autoimmune Diseases; Autoimmunity; B-Lymphocytes; Benchmarking; Biogenesis; Biological Markers; Blood Circulation; Body Fluids; Cataloging; Catalogs; Cells; Clinical; Collaborations; Complex; Core Facility; Cytoplasmic Granules; DNA; Deposition; Development; Disease; Endoribonucleases; Family member; Foundations; Future; Gender; Harvest; Health; Homeostasis; Human; Immune; Immune response; Immunity; Immunologic Receptors; Inclusion Bodies; Individual; Lead; Mammals; Mediating; Modeling; Molecular; Monitor; Mutation; Myopathy; Natural Immunity; Neurodegenerative Disorders; Patients; Pattern recognition receptor; Play; Process; Proteins; RNA; RNA Binding; Race; Recording of previous events; Regulation; Research; Research Personnel; Research Proposals; Rheumatism; Ribonucleoproteins; Role; Sampling; Serum; Signal Transduction; Solid; Specificity; Stress; Structure; System; Systemic Lupus Erythematosus; Transfer RNA; base; biological adaptation to stress; clinical material; endoribonuclease; established cell line; extracellular; genome sequencing; in vivo; novel; pathogen; peptide hormone; prion-like; transcriptome sequencing

DESCRIPTION (provided by applicant): Extracellular RNA (exRNA) from pathogens plays a key role as activator of innate immunity in mammals. However, the presence of host exRNA in serum and other body fluids challenges the current models of RNA based immune recognition. To understand its basis of specificity, it is important to catalog host exRNAs and their associated proteins in serum, investigate mechanisms leading to circulating ribonucleoprotein (RNP) homeostasis, and identify protein factors contributing to cellular RNA release. Genetic alterations in RNA targets or interacting proteins may contribute to imbalances in normal versus stress-triggered release of RNPs and push adaptive long-term pathogen-directed immunity towards autoimmunity. ExRNAs may also play a broader role in extracellular signaling similar to peptide hormones, which would also be captured by this experimental approach. This application brings together a team of multiple investigators with complementary expertise and history of close collaboration to build a solid foundation regarding identification, mechanism and function of extracellular RNAs and the proteins involved in their biogenesis, export, recognition, and turnover. The specific aims of the proposed project are: 1. Catalogue and quantify all classes of extracellular RNAs in human serum from normal subjects using various established and novel RNA seq approaches and examine normal variability of circulating RNA profiles within an individual, between individuals and the influence of gender, age, race, and disease. Establish a core facility for processing and archiving clinical materials (Williams, Putterman, Tuschi). 2. Determine exRNA composition in patients suffering from systemic lupus erythematosus (SLE), for whom antibodies against different classes of RBPs is a hallmark. Considering that these RBPs alter their subcellular localization upon stress and appear in stress granules with immature RNA and/or RNA targeted for turnover, we will evaluate in as much the composition of RNPs in stress granules harvested from immortalized B cells of normal and SLE subjects possesses immunostimulatory function and if these RNP granules are also released during stress (Tuschi, Putterman, Williams). 3. Develop a molecular and mechanistic understanding of stress granule formation and RNA/RNP mediated innate immune responses. Identify the RNA targets and RNP structures of autoantigens in tRNA stress responses, their turnover, and their immune receptors.

描述（由申请人提供）： 病原体的细胞外RNA（EXRNA）作为哺乳动物先天免疫的活化剂起着关键作用。然而，血清和其他体液中宿主EXRNA的存在挑战了基于RNA的免疫识别的当前模型。为了了解其特异性的基础，对于血清中宿主EXRNA及其相关蛋白的目录非常重要，研究导致循环核糖核蛋白（RNP）稳态的机制，并鉴定有助于细胞RNA释放的蛋白质因子。 RNA靶标或相互作用蛋白的遗传改变可能导致正常与应力触发的RNP释放的失衡，并将适应性的长期病原体指导的免疫力推向自身免疫性。 EXRNA在类似于肽激素的细胞外信号传导中也可能发挥更广泛的作用，这种实验方法也将捕获。该应用程序汇集了一个具有互补专业知识和密切合作历史的多个研究人员的团队，以建立有关细胞外RNA的识别，机制和功能的坚实基础，以及与其生物发生，出口，识别和营业额有关的蛋白质。拟议项目的具体目的是：1。使用各种已建立和新颖的RNA SEQ方法从正常受试者中的人血清中的所有类别的细胞外RNA进行量化，并检查个体，性别，年龄，种族，种族和疾病的个体内部循环RNA特征的正常变异性。建立用于处理和归档临床材料的核心设施（Williams，Putterman，Tuschi）。 2。确定患有全身性红斑狼疮（SLE）患者的EXRNA组成，针对不同类别的RBP的抗体是标志。考虑到这些RBP在压力时改变了它们的亚细胞定位，并出现在未成熟的RNA和/或RNA的应力颗粒中，我们将在正常和SLE受试者中具有永生的B细胞中的不良B细胞中收获的RNP的组成中评估RNP的大量成分，并具有免疫性功能，如果这些RNP GRANES也释放出了（在此期间）。 3。对应激颗粒形成和RNA/RNP介导的先天免疫反应的分子和机械理解。识别自身抗原在tRNA应力反应，其周转率和免疫受体中的RNA靶标和RNP结构。