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: 9450828
• 负责人: THOMAS TUSCHL
• 金额: $ 7.71万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9450828
• 关键词: Affect; Age; Amyloid; Antibodies; Archives; Autoantigens; Autoimmune Diseases; Autoimmunity; B lymphocyte immortalization; Benchmarking; Biogenesis; Biological Markers; Blood Circulation; Body Fluids; Catalogs; Cells; Clinical; Collaborations; Complex; Core Facility; Cytoplasmic Granules; DNA; Deposition; Development; Disease; Endoribonucleases; Family member; Foundations; Future; Gender; Harvest; Health; Homeostasis; Human; Immune; Immunity; Immunologic Receptors; Inclusion Bodies; Individual; Innate Immune Response; Lead; Mammals; Mediating; Modeling; Molecular; Monitor; Mutation; Myopathy; Natural Immunity; Neurodegenerative Disorders; Patients; Pattern recognition receptor; Play; 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; public health relevance; 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)： 病原体产生的细胞外RNA(ExRNA)作为哺乳动物天然免疫的激活剂起着关键作用。然而，血清和其他体液中宿主exRNA的存在对目前基于RNA的免疫识别模型提出了挑战。为了了解其特异性的基础，重要的是对血清中宿主exRNAs及其相关蛋白进行分类，研究导致循环核糖核蛋白(RNP)动态平衡的机制，并确定促进细胞RNA释放的蛋白质因素。RNA靶标或相互作用蛋白的遗传改变可能导致正常和应激触发的RNPs释放的失衡，并将适应性长期病原体定向免疫推向自身免疫。ExRNAs也可能在细胞外信号传递中发挥更广泛的作用，类似于多肽激素，这也将被这种实验方法捕获。这一应用将具有互补专业知识和密切合作历史的多个研究人员团队聚集在一起，以建立关于细胞外RNA及其生物发生、输出、识别和周转所涉及的蛋白质的识别、机制和功能的坚实基础。拟议项目的具体目标是：1.使用各种已建立的和新的RNA序列方法，对正常人血清中的所有类别的细胞外RNA进行编目和量化，并检查个人内部、个人之间循环RNA图谱的正常变异性以及性别、年龄、种族和疾病的影响。建立处理和归档临床材料的核心设施(威廉姆斯、普特曼、图斯基)。2.检测系统性红斑狼疮(SLE)患者外显性RNA的组成，以抗不同类型RBPs抗体为标志。考虑到这些RNP在应激时改变了它们的亚细胞定位，并以未成熟的RNA和/或RNA为目标出现在应激颗粒中，我们将评估从正常人和SLE受试者的永生化B细胞中获取的应激颗粒中RNP的组成是否具有免疫刺激功能，以及这些RNP颗粒是否在应激期间也被释放(Tuschi，Putterman，Williams)。3.从分子和机制上理解应激颗粒的形成和RNA/RNP介导的先天免疫反应。确定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。

项目成果

International validation of a urinary biomarker panel for identification of active lupus nephritis in children.

• DOI: 10.1007/s00467-016-3485-3
• 发表时间: 2017-02
• 期刊: Pediatric nephrology (Berlin, Germany)
• 作者: Smith EM;Jorgensen AL;Midgley A;Oni L;Goilav B;Putterman C;Wahezi D;Rubinstein T;Ekdawy D;Corkhill R;Jones CA;Marks SD;Newland P;Pilkington C;Tullus K;Beresford MW
• 通讯作者: Beresford MW

Structure-based insights into self-cleavage by a four-way junctional twister-sister ribozyme.

基于结构的四路连接扭转姐妹核酶自裂解的见解

• DOI: 10.1038/s41467-017-01276-y
• 发表时间: 2017-10-30
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Zheng L;Mairhofer E;Teplova M;Zhang Y;Ma J;Patel DJ;Micura R;Ren A
• 通讯作者: Ren A

Structure-based mechanistic insights into catalysis by small self-cleaving ribozymes.

基于结构的小型自裂解核酶催化机制的见解

• DOI: 10.1016/j.cbpa.2017.09.017
• 发表时间: 2017-12
• 期刊: Current opinion in chemical biology
• 影响因子: 7.8
• 作者: Ren A;Micura R;Patel DJ
• 通讯作者: Patel DJ

A Mini-Twister Variant and Impact of Residues/Cations on the Phosphodiester Cleavage of this Ribozyme Class.

• DOI: 10.1002/anie.201506601
• 发表时间: 2015-12-07
• 期刊: Angewandte Chemie (International ed. in English)
• 作者: Košutić M;Neuner S;Ren A;Flür S;Wunderlich C;Mairhofer E;Vušurović N;Seikowski J;Breuker K;Höbartner C;Patel DJ;Kreutz C;Micura R
• 通讯作者: Micura R

c-di-AMP binds the ydaO riboswitch in two pseudo-symmetry-related pockets.

• DOI: 10.1038/nchembio.1606
• 发表时间: 2014-09
• 期刊: NATURE CHEMICAL BIOLOGY
• 影响因子: 14.8
• 作者: Ren, Aiming;Patel, Dinshaw J.
• 通讯作者: Patel, Dinshaw J.