Expression and function of microRNA in autoimmune Sjogren's Syndrome

microRNA在自身免疫性干燥综合征中的表达和功能

基本信息

批准号：
8668761
负责人：
SEUNGHEE CHA
金额：
$ 36.26万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8668761
关键词：
3&apos Untranslated Regions Acids Address Affect Age American Autoimmune Diseases Autoimmune Process Autoimmunity Biological Biological Assay Biological Markers Biopsy Specimen Cell Line Cells Characteristics Clinical Complex Computer Simulation Control Groups Data Development Diagnosis Diagnostic Disease Disease Marker Epigenetic Process Exhibits Exocrine Glands Female Functional RNA Functional disorder Gene Targeting Genes Goals Human Immune Immune response In Situ Hybridization In Vitro Individual Inflammation Inflammatory Lacrimal gland structure Luciferases MicroRNAs Natural Immunity Nature Nuclear Onset of illness Pathogenesis Patients Peripheral Blood Mononuclear Cell Phagocytosis Play Prevention Process Production Quality of life RNA Interference Recruitment Activity Regulation Research Research Design Research Proposals Rheumatoid Arthritis Role Salivary Glands Screening Result Sjogren&apos s Syndrome Specificity Staging Stimulus Syndrome System Systemic Lupus Erythematosus Testing Therapeutic Therapeutic Intervention Time Tissues Transfection Up-Regulation Viral Xerostomia cytokine eye dryness human old age (65+)improved in vitro testing in vivo inhibitor/antagonist insight monocyte mouse model novel research study response screening tool

项目摘要

DESCRIPTION (provided by applicant): Autoimmune Sj"gren's syndrome (SjS) targets the exocrine glands, such as the salivary and lacrimal glands, of mostly female individuals, leading to severe secretory dysfunction and its complications. To date, SjS- specific diagnostics and therapeutics have not been available due to complexity of SjS disease pathogenesis. MicroRNAs (miRNAs) have been recognized as an important class of non-coding RNAs that are involved in a variety of biological, pathological, and immunological processes. Our latest findings on altered miR-146a and- 155 expression in peripheral blood mononuclear cells (PBMC) or monocytes of primary SjS (pSjS) patients in comparison with healthy controls indicate that miRNAs may play a critical role in autoimmune initiation and progression of SjS. The data were further supported by the findings in the salivary glands and PBMC of our pSjS-prone C57BL/6.NOD-Aec1Aec2 mouse model, where up-regulation of those miRNAs was observed from an early age on. Over-expression of miR-146a in human monocyte cell line (THP-1) was able to alter innate immune responses such as phagocytosis and pro-inflammatory cytokine production in our preliminary study in vitro. We now propose studies designed to test our hypotheses that miR-146a and miR-155 are vital to prolong inflammatory processes by altering innate immune response and that pSjS-specific miRNAs especially involved in innate immunity can be differentially expressed (potentially even prior to disease onset resulting in persistent inflammation). In Aim 1 of this project, we will investigate consequences of altered miR-146a and/or miR-155 by utilizing miRNA mimics and inhibitors transfected into THP-1 cells followed by functional assays. Altered functions will also be tested in primary cultured monocytes from pSjS patients. This aim is to determine if epigenetic regulation of immune genes by miRNAs are critical in altering immunological processes in SjS. In Aim 2, we will begin to profile a complex network of miRNAs involved in pSiS by utilizing a miRNA microarray approach. Here we will test our hypothesis that miRNA profiles that are specific for human pSjS are identifiable. Furthermore, we will continue to identify in Aim 2 target genes of differentially expressed miRNAs and perform functional assays established in Aim 1 to screen their involvement in innate immune responses. We envision that these experiments will identify novel miRNAs in pSjS and regulatory functions of identified miRNAs in abnormal immune regulation of SjS and may ultimately be amenable to development of biomarkers and therapeutic intervention.

描述（由申请人提供）：自身免疫性干燥综合征（SjS）主要针对女性个体的外分泌腺，例如唾液腺和泪腺，导致严重的分泌功能障碍及其并发症。迄今为止，SjS特异性诊断和治疗由于 SjS 疾病发病机制的复杂性，MicroRNA (miRNA) 已被认为是一类重要的非编码 RNA，参与多种疾病的治疗。我们对原发性 SjS (pSjS) 患者外周血单核细胞 (PBMC) 或单核细胞 (pSjS) 与健康对照相比 miR-146a 和 155 表达改变的最新发现表明，miRNA 可能发挥着关键作用。在 SjS 的自身免疫启动和进展中的作用这些数据得到了 pSjS 易感者唾液腺和 PBMC 的研究结果的进一步支持。 C57BL/6.NOD-Aec1Aec2 小鼠模型，从很小的时候就观察到这些 miRNA 的上调。在我们的体外初步研究中，人单核细胞系 (THP-1) 中 miR-146a 的过度表达能够改变先天免疫反应，例如吞噬作用和促炎细胞因子的产生。我们现在提出旨在测试我们假设的研究，即 miR-146a 和 miR-155 通过改变先天免疫反应对于延长炎症过程至关重要，并且 pSjS 特异性 miRNA 特别是参与先天免疫的 miRNA 可以差异表达（甚至可能在疾病发作之前）导致持续性炎症）。在该项目的目标 1 中，我们将通过利用转染至 THP-1 细胞的 miRNA 模拟物和抑制剂，然后进行功能测定来研究 miR-146a 和/或 miR-155 改变的后果。还将在 pSjS 患者的原代培养单核细胞中测试改变的功能。该目的是确定 miRNA 对免疫基因的表观遗传调控对于改变 SjS 的免疫过程是否至关重要。在目标 2 中，我们将利用 miRNA 微阵列方法开始分析 pSiS 中涉及的复杂 miRNA 网络。在这里，我们将检验我们的假设，即人类 pSjS 特有的 miRNA 图谱是可识别的。此外，我们将继续在 Aim 2 中鉴定差异表达 miRNA 的靶基因，并进行 Aim 1 中建立的功能测定，以筛选它们在先天免疫反应中的参与。我们设想这些实验将鉴定 pSjS 中的新 miRNA 以及所鉴定的 miRNA 在 SjS 异常免疫调节中的调节功能，并可能最终适合生物标志物的开发和治疗干预。