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

• 批准号: 8100296
• 负责人: SEUNGHEE CHA
• 金额: $ 35.53万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8100296
• 关键词: 3' 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; Screening procedure; Sjogren' 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; public health relevance; research study; response; tool

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. PUBLIC HEALTH RELEVANCE: Sj"gren's syndrome (SjS) affects 4 million Americans, resulting in dry mouth and dry eye condition in patients. Our research proposal to identify SjS-specific profiles would advance diagnostics and therapeutics in the field and improve quality of life in those affected with the disease.

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，由于 SjS 疾病发病机制的复杂性，尚未有针对 SjS 的特异性诊断和治疗方法。MicroRNA（miRNA）已得到认可 作为一类重要的非编码RNA，参与多种生物学、病理学和免疫学过程。我们关于原发性 SjS (pSjS) 患者的外周血单核细胞 (PBMC) 或单核细胞 (pSjS) 与健康对照相比的 miR-146a 和 155 表达改变的最新发现表明，miRNA 可能在 SjS 的自身免疫启动和进展中发挥关键作用。的 我们的 pSjS 倾向 C57BL/6.NOD-Aec1Aec2 小鼠模型的唾液腺和 PBMC 中的发现进一步支持了这些数据，其中从很小的时候就观察到这些 miRNA 的上调。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 在我们的体外初步研究中。我们现在提出的研究旨在检验我们的假设，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，并且特别参与先天免疫的 pSjS 特异性 miRNA 可以差异表达（甚至可能在导致持续炎症的疾病发作之前）。在该项目的目标 1 中，我们将调查更改的后果 miR-146a 和/或 miR-155 通过利用 miRNA 模拟物和抑制剂转染到 THP-1 细胞中，然后进行功能测定。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，我们将开始分析所涉及的 miRNA 的复杂网络 通过利用 miRNA 微阵列方法在 pSiS 中进行。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 以及已鉴定的调节功能 miRNA 参与 SjS 的异常免疫调节，最终可能适合生物标志物的开发和治疗干预。 公共健康相关性：干燥综合征 (SjS) 影响着 400 万美国人，导致患者出现口干和眼干症状。我们的研究计划旨在确定 SjS 的特异性特征，这将推进该领域的诊断和治疗，并改善该疾病患者的生活质量。