Micro-RNAs as biomarkers and targets of herbal CAM in autoimmune arthritis.

Micro-RNA 作为自身免疫性关节炎中草药 CAM 的生物标志物和靶标。

• 批准号: 9396022
• 负责人: Steven Dudics
• 金额: $ 4.4万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-15 至 2019-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9396022
• 关键词: Adjuvant Arthritis; Adverse effects; Adverse reactions; Affect; Alpha Cell; American; Antigens; Arthritis; Autoantigens; Autoimmune Diseases; Autoimmune Process; Autoimmune Responses; Base Pairing; Binding; Biological Markers; Celastrus; Cell Line; Cells; Chinese Herbs; Chinese Traditional Medicine; Chondrocytes; Chronic; Collagen Type II; Complementary and alternative medicine; Country; Data; Degenerative polyarthritis; Development; Diagnostic; Disease; Early Diagnosis; Endoribonucleases; Environmental Risk Factor; Etiology; Fibroblasts; Gene Expression; Gene Targeting; Genes; Genetic; Heat shock proteins; Herbal Medicine; Human; Immunochemistry; Immunosuppression; Infection; Inflammation Mediators; Inflammatory; Inflammatory Arthritis; Joints; Longevity; Malignant Neoplasms; Mediating; Medical; Medicinal Herbs; Messenger RNA; MicroRNAs; Modeling; Monitor; Natural Products; Nucleotides; Organ; Pathogenesis; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phase; Plants; Play; Population; Predisposition; Process; Quantitative Reverse Transcriptase PCR; RNA; RNA Sequences; Rattus; Regulator Genes; Research Personnel; Rheumatoid Arthritis; Role; Self Tolerance; TNF gene; Therapeutic; Tissues; Translations; Untranslated RNA; Western Blotting; Work; arthritis therapy; autoimmune arthritis; base; differential expression; disease diagnosis; drug development; gene function; improved; interest; lymph nodes; novel therapeutics; prevent; tool; tripterine

Project Summary Rheumatoid arthritis (RA) is a multifactorial disease that involves both a genetic and an environmental component. Approximately 0.5-1% of the population in the U.S. is afflicted with RA. Current treatments, including biologics such as anti-tumor necrosis factor-α, are effective, but only in about 50-60% of RA patients. In addition, their use is associated with severe adverse effects, such as immune suppression that renders RA patients more vulnerable to infections. In order to develop better therapeutics, a more comprehensive understanding of the pathogenesis of RA is required. Our lab has previously shown that there is a dynamic change in gene expression at different phases of adjuvant-induced arthritis (AA) in Lewis rats. Furthermore, a traditional Chinese medicine celastrus and its bioactive component celastrol can suppress AA. However, their mechanisms of action have not been fully defined. Micro-RNAs (miRNAs) are short (~19-22 nucleotides long), non-coding RNA sequences that repress the activity of specific genes. These short sequences either inhibit translation or cause degradation of the target messenger RNA (mRNA). We hypothesize that miRNAs play a vital role in the development and progression of autoimmune arthritis, and that specific miRNAs can also serve as biomarkers for this disease. Moreover, we suggest that celastrus/celastrol mediates its effect in part by regulating the levels of specific miRNAs. Aim 1. To define the miRNA profiles of arthritic Lewis rats at early and late phases of AA, and identify significantly altered and relevant miRNAs as well as their potential target genes that are involved in arthritis pathogenesis. Aim 2. To identify specific miRNAs whose expression is significantly altered following treatment of arthritic rats with celastrus or its bioactive component celastrol, and to define the role of those miRNAs in mediating the anti-arthritic activity of these herbal complementary and alternative medicine (CAM) products. Using total RNA purified from the draining lymph node cells and the synovial-infiltrating cells of Lewis rats following an arthritogenic challenge, with or without treatment with celastrus/celastrol, we plan to use miRNA microarray to identify miRNAs of interest. To further validate the selected miRNA, we will transfect a cell line with miRNA mimic/antagonist to over-/under-express, respectively the miRNA of interest. Then we will employ qRT-PCR, western blotting, and immunochemistry to determine their effects on target genes associated with arthritis pathogenesis as well as their control by herbal CAM therapy. We believe that our results would not only advance the understanding of arthritis pathogenesis, but also help develop novel therapeutics for RA.

项目摘要 类风湿关节炎（RA）是一种多因素疾病，涉及遗传和环境 成分。美国大约0.5-1％的人口遭受了RA的折磨。当前治疗， 包括抗肿瘤坏死因子-α等生物制剂有效，但仅在50-60％的RA患者中。 此外，它们的使用与严重的不良反应有关，例如免疫抑制作用，使RA导致RA 患者更容易感染。为了开发更好的治疗剂，更全面 需要理解RA的发病机理。我们的实验室以前已经表明有动态 在刘易斯大鼠的调整诱导关节炎（AA）的不同阶段，基因表达的变化。此外， 中药及其生物活性成分Celastrul可以抑制AA。但是，他们 作用机制尚未完全定义。微RNA（miRNA）短（长19-22个核苷酸）， 反映特定基因活性的非编码RNA序列。这些短序列可以抑制 转换或导致目标信使RNA（mRNA）的降解。我们假设mirnas扮演 在自身免疫性关节炎的发展和发展中的重要作用，并且特定的miRNA也可以服务 作为这种疾病的生物标志物。此外，我们建议celastrus/celastrol培养其影响部分 调节特定miRNA的水平。目的1。在早期定义关节性刘易斯大鼠的miRNA特征 和AA的后期阶段，并确定明显改变和相关的miRNA及其潜在目标 参与关节炎发病机理的基因。目标2。确定表达的特定miRNA 用Celastrus或其生物活性成分celastrol治疗关节大鼠后，显着改变了 定义这些miRNA在介导这些草药完整性和 替代药物（CAM）产品。使用从排水淋巴结细胞纯化的总RNA和 在关节炎挑战之后，刘易斯大鼠的滑膜渗透细胞，或不接受治疗 Celastrous/celastrol，我们计划使用miRNA微阵列来识别感兴趣的miRNA。进一步验证 选定的miRNA，我们将分别用miRNA模拟/拮抗剂将细胞系转换为过表现 感兴趣的mirna。然后，我们将采用QRT-PCR，蛋白质印迹和免疫化学来确定 它们对与关节炎发病机理相关的靶基因以及草药凸轮控制的影响 治疗。我们认为，我们的结果不仅会推动对关节炎发病机理的理解，还会提高 还可以帮助开发RA的新型疗法。