MicroRNA-based therapy for rheumatoid arthritis

基于 MicroRNA 的类风湿性关节炎疗法

基本信息

批准号：
10475349
负责人：
Salah-uddin Ahmed
金额：
$ 15.3万
依托单位：
WASHINGTON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-12 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10475349
关键词：
Adjuvant Adjuvant Arthritis Antiinflammatory Effect Biogenesis Biotin Clinical Data Disease Enzymes Epigenetic Process Exoribonucleases Fibroblasts Genes Genetic Transcription Human IL8 gene In Vitro Incidence Inflammation Interleukin-1 beta Interleukin-6 Interstitial Collagenase Joints Knowledge Label Link Malignant Neoplasms Methods MicroRNAs Modeling Molecular Mus Pathogenesis Pathway Analysis Play Polyubiquitination Process Production RNA Editing Rattus Regulation Replacement Therapy Rheumatoid Arthritis Ribonucleases Role Sampling Serum Serum Markers Severities Signal Pathway Signal Transduction Signal Transduction Pathway System TNF gene TRAF2 gene Testing Therapeutic Tissues Transferase Transgenic Organisms Ubiquitin Untranslated RNA Validation base cell type collagenase 3 cytokine efficacy evaluation efficacy testing efficacy validation functional restoration human model in vivo in vivo evaluation molecular phenotype mouse model multicatalytic endopeptidase complex novel novel therapeutics restoration success therapeutic miRNA transcriptome sequencing ubiquitin-protein ligase uridylate

项目摘要

PROJECT SUMMARY MicroRNAs (miRNAs) are single stranded non-coding RNAs that have emerged as a novel posttranscriptional regulators in rheumatoid arthritis (RA) pathogenesis. However, recent studies provide evidence that in general there is an overall reduction in miRNA expression in disease states such as cancer and RA. While the rationale for developing miRNA therapeutics are conceptually similar to other inhibitory approaches, restoring the function of a miRNA by miRNA replacement is a less characterized, yet potential therapeutic option never tested in RA. Intriguingly, the underlying reasons for disrupted miRNA biogenesis and degradation processes in RA are not fully understood. Our recent finding provides evidence that miR-17 expression was significantly low in RA serum, synovial fibroblasts (SFs), and synovial tissues (STs), as well as in the serum and joints of adjuvant-induced (AIA) rats. RNA-sequencing analysis showed modulation of 664 genes by the restoration of miR-17 levels using precursor (pre)-miR-17 in human RASFs. Ingenuity pathway analysis of RNA-sequencing data identified the ubiquitin proteasome system in the TNF-α signaling pathway as a primary target of miR-17. Furthermore, the restoration of miR-17 levels using precursor-miR- 17 (pre-miR-17) reduced the ability of ubiquitin E3 ligase TRAF2 to associate with its signaling partner, cIAP2, thereby inhibiting TNF-α-induced downstream signal transduction pathways and suppressing the production of IL-6, IL-8, MMP-1, and MMP-13 in human RASFs. While these novel findings provide evidence for the impact of miR-17 replacement on posttranslational processes critical in TNF-α signaling in RASFs, several questions important to miR- 17 biogenesis and turnover remains unanswered, including the reasons for severely low expression in RA, the impact of proinflammatory cytokines on miR-17 biogenesis and turnover, and the relevance and efficacy of miR-17 replacement therapy in RA. Based on these novel observations, we propose that miR-17 replacement therapy could ameliorate RA. Thus, in specific aim 1, we will determine the deregulated mechanism of miR-17 biogenesis and turnover in RA. In aim 2, we will evaluate the relevance and molecular mechanisms of miR-17 restoration on TNF-α- induced molecular and phenotypic changes in human RASFs. Finally, aim 3 will test the in vivo efficacy of miR-17 delivery in TNF-α transgenic (hTNF-tg) mouse model and rat AIA model of human RA. The success of these studies will lead to two clinically distinct findings: 1) Elucidation of the altered miRNA biogenesis and turnover mechanism in RA pathogenesis, and 2) the validation of miRNA-based therapeutic approaches for the treatment of RA.

项目概要 MicroRNA (miRNA) 是单链非编码 RNA，已作为一种新型转录后RNA 出现。然而，最近的研究提供的证据表明，总体上存在类风湿性关节炎（RA）发病机制中的调节因子。是癌症和 RA 等疾病状态下 miRNA 表达的总体减少。开发 miRNA 疗法在概念上与其他抑制方法类似，恢复 miRNA 的功能 miRNA 替代 miRNA 的特征较少，但从未在 RA 中进行过测试。 RA 中 miRNA 生物发生和降解过程被破坏的根本原因尚不完全清楚。我们最近的发现提供了证据表明 RA 血清、滑膜成纤维细胞中 miR-17 表达显着较低 (SF)、滑膜组织 (ST) 以及佐剂诱导 (AIA) 大鼠的血清和关节中。分析显示，在人类中使用前体 (pre)-miR-17 可通过恢复 miR-17 水平来调节 664 个基因 RNA 测序数据的 RASF 通路分析确定了 TNF-α 中的泛素蛋白酶体系统。信号通路作为 miR-17 的主要靶标此外，使用前体-miR-恢复 miR-17 水平。 17 (pre-miR-17) 降低了泛素 E3 连接酶 TRAF2 与其信号传导伙伴 cIAP2 关联的能力，从而抑制TNF-α诱导的下游信号转导途径并抑制IL-6、IL-8、人类 RASF 中的 MMP-1 和 MMP-13 而这些新发现为 miR-17 的影响提供了证据。 RASF 中对 TNF-α 信号转导至关重要的翻译后过程的替代，是 miR- 的几个重要问题 17 生物发生和周转仍未得到解答，包括 RA 中严重低表达的原因、影响促炎细胞因子对 miR-17 生物发生和周转的影响，以及 miR-17 的相关性和功效基于这些新的观察结果，我们提出 miR-17 替代疗法可以治疗 RA。因此，在具体目标 1 中，我们将确定 miR-17 生物合成的失调机制和在目标 2 中，我们将评估 miR-17 恢复对 TNF-α- 的相关性和分子机制。最后，目标 3 将测试 miR-17 的体内功效。这些研究在 TNF-α 转基因 (hTNF-tg) 小鼠模型和人类 RA 大鼠 AIA 模型中的递送取得了成功。将导致两个临床上不同的发现：1) 阐明改变的 miRNA 生物发生和周转机制 RA 发病机制，2) 验证基于 miRNA 的 RA 治疗方法。