Elucidating a role for a hematopoietic-tropic microRNA in autoreactive T cells

阐明促造血 microRNA 在自身反应性 T 细胞中的作用

基本信息

批准号：
8285550
负责人：
ANTOINE L. MENORET
金额：
$ 23.1万
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：
2012
资助国家：
美国
起止时间：
2012-03-02 至 2014-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8285550
关键词：
Ablation Adjuvant Adopted Adoptive Transfer Adverse effects Affect Anti-Inflammatory Agents Anti-inflammatory Applications Grants Area Autoimmune Diseases Autoimmune Process Autoimmunity Binding Binding Sites Bioinformatics Biological Assay CD4 Positive T Lymphocytes CD8B1 gene Cell physiology Cells Characteristics Cloning Complementary DNA Data Detection Developmental Biology Disease Disease model Effector Cell Enzymes Experimental Autoimmune Encephalomyelitis Firefly Luciferases Functional RNA Gene Expression Gene Targeting Generations Genes Genetic Health Hematopoietic Human Immune Immune response Immune system Inflammation Lead Libraries Luciferases Lymphoid Tissue Maps Mediating Messenger RNA MicroRNAs Modeling Molecular Molecular Conformation Monitor Multiple Sclerosis Mus Myelin Basic Proteins Northern Blotting Nucleotides Outcome Pathway interactions Patients Physiological Play Proteins Proteome Proteomics RNA RNA Processing Regulation Research Research Personnel Role Small RNA Sum T cell response T-Cell Activation T-Cell Activation Pathway T-Lymphocyte Technology Testing Therapeutic Tissues Transactivation Transcript Translational Repression Translations Variant Work autoreactive T cell cDNA Library clinically relevant cytokine experience hematopoietic tissue human DICER1 protein immune activation immune function improved in vivo insight interest mRNA Stability mouse model novel outcome forecast prevent response septic stem synergism tool translational approach vector

项目摘要

DESCRIPTION (provided by applicant): The use of microRNA's (miRNA) for the study, detection, prognosis and cure of disease is growing and becoming a reliable tool toward improving human health. In this regard the immune system is an important area since T cells are greatly impacted by deletion of dicer a key RNA processing enzyme required for the generation of miRNA. To gain new insight into how miRNA impact T cell responses, mice were immunized with costimulation and adjuvant to generate potent T effector responses. We generated a cDNA library that could trace changes in miRNA derived from lymphoid tissue. Using sequencing of size-fractionated cDNA we cloned 6 potential miRNAs from this library and found candidates that possessed characteristic stem loop folding and were dicer dependent. One of these candidates, termed miR-R89, was unique compared to the others since its expression was hematopoietic specific and also regulated during immune activation. The targets and function of miR-R89 are currently unknown but our new preliminary data show that it is expressed in a T cell clone known to cause experimental autoimmune encephalitis (EAE). In Aim 1 we will use a powerful proteomic strategy to examine the effects of blocking miR-R89 on the T cell proteome followed by our bioinformatics gene targeting data and a molecular approach to validate potential target genes. In Aim 2 we will study how miR-R89 inhibition affects the initiation and progression of EAE. Thus, this R21 proposal will explore how a new miRNA can impact specific T cell function, which may lead to a novel translational approach for multiple sclerosis in humans. PUBLIC HEALTH RELEVANCE: This application will characterize the immune and therapeutic potential of a small RNA we cloned, which may be able to control gene expression of important pathways that impact immune T cell activation. This notion will be tested in a mouse model of Multiple Sclerosis (MS) with the hope of developing more efficient therapy for MS patients. Our work will bring fresh insight into the field and add greatly to the control of immune gene expression in the immune system by microRNA.

描述（由申请人提供）：微小RNA（miRNA）在疾病研究、检测、预测和治疗中的应用正在不断增长，并成为改善人类健康的可靠工具。在这方面，免疫系统是一个重要领域，因为 T 细胞会受到 dicer（生成 miRNA 所需的关键 RNA 加工酶）缺失的极大影响。为了获得关于 miRNA 如何影响 T 细胞反应的新见解，用共刺激和佐剂对小鼠进行免疫，以产生有效的 T 效应反应。我们生成了一个 cDNA 文库，可以追踪淋巴组织中 miRNA 的变化。通过对按大小分级的 cDNA 进行测序，我们从该文库中克隆了 6 个潜在的 miRNA，并发现了具有特征性茎环折叠且依赖切丁机的候选物。其中一种候选物被称为 miR-R89，与其他候选物相比是独特的，因为它的表达具有造血特异性，并且在免疫激活过程中也受到调节。 miR-R89 的靶标和功能目前尚不清楚，但我们新的初步数据表明，它在已知引起实验性自身免疫性脑炎 (EAE) 的 T 细胞克隆中表达。在目标 1 中，我们将使用强大的蛋白质组策略来检查阻断 miR-R89 对 T 细胞蛋白质组的影响，然后使用我们的生物信息学基因靶向数据和分子方法来验证潜在的靶基因。在目标 2 中，我们将研究 miR-R89 抑制如何影响 EAE 的发生和进展。因此，这个 R21 提案将探索新的 miRNA 如何影响特定的 T 细胞功能，这可能会为人类多发性硬化症带来一种新的转化方法。公共健康相关性：该应用将表征我们克隆的小 RNA 的免疫和治疗潜力，它可能能够控制影响免疫 T 细胞激活的重要途径的基因表达。这一概念将在多发性硬化症（MS）小鼠模型中进行测试，希望为多发性硬化症患者开发更有效的治疗方法。我们的工作将为该领域带来新的见解，并极大地增强 microRNA 对免疫系统中免疫基因表达的控制。