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: 9124822
• 负责人: THOMAS TUSCHL
• 金额: $ 134.69万
• 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 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9124822
• 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; 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）： 来自病原体的细胞外 RNA (exRNA) 作为哺乳动物先天免疫的激活剂发挥着关键作用。然而，血清和其他体液中宿主 exRNA 的存在对当前基于 RNA 的免疫识别模型提出了挑战。为了了解其特异性的基础，重要的是对血清中的宿主 exRNA 及其相关蛋白进行分类，研究导致循环核糖核蛋白 (RNP) 稳态的机制，并识别有助于细胞 RNA 释放的蛋白质因子。 RNA靶标或相互作用蛋白的基因改变可能导致正常与应激触发的RNP释放失衡，并推动适应性长期病原体定向免疫发展为自身免疫。 ExRNA 也可能在类似于肽激素的细胞外信号传导中发挥更广泛的作用，这一实验方法也将捕获这一点。该应用汇集了多个研究人员组成的团队，他们具有互补的专业知识和密切合作的历史，为细胞外 RNA 以及参与其生物发生、输出、识别和周转的蛋白质的鉴定、机制和功能奠定了坚实的基础。该项目的具体目标是： 1. 使用各种已建立的和新颖的 RNA seq 方法对正常受试者人血清中所有类别的细胞外 RNA 进行编目和量化，并检查个体内、个体之间循环 RNA 谱的正常变异性及其影响性别、年龄、种族和疾病。建立处理和归档临床材料的核心设施（Williams、Putterman、Tuschi）。 2. 确定系统性红斑狼疮 (SLE) 患者的 exRNA 组成，针对不同类别 RBP 的抗体是该患者的标志。考虑到这些 RBP 在应激时会改变其亚细胞定位，并出现在具有未成熟 RNA 和/或针对周转的 RNA 的应激颗粒中，我们将尽可能多地评估从正常和 SLE 受试者的永生化 B 细胞中收获的应激颗粒中的 RNP 组成。免疫刺激功能，以及这些 RNP 颗粒是否在应激期间也被释放（Tuschi、Putterman、Williams）。 3. 建立对应激颗粒形成和 RNA/RNP 介导的先天免疫反应的分子和机制理解。识别 tRNA 应激反应中自身抗原的 RNA 靶标和 RNP 结构、其周转及其免疫受体。