MicroRNA-based therapy for rheumatoid arthritis

基于 MicroRNA 的类风湿性关节炎疗法

• 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
• 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

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.

项目总结 MicroRNAs(MiRNAs)是作为一种新的转录后转录因子而出现的单链非编码RNA 类风湿关节炎(RA)发病机制中的调节因子。然而，最近的研究提供的证据表明，总的来说 是癌症和类风湿性关节炎等疾病状态下miRNA表达的总体减少。虽然这样做的理由 开发miRNA疗法在概念上类似于其他抑制方法，恢复 通过miRNA替代的miRNA是一种特征较少但潜在的治疗方案，从未在RA中进行过试验。有趣的是， RA中miRNA生物发生和降解过程中断的潜在原因尚不完全清楚。 我们最近的发现提供了证据，表明miR-17在RA血清、滑膜成纤维细胞中的表达显著降低 (SFS)和滑膜组织(STS)，以及佐剂诱导(AIA)大鼠的血清和关节中。RNA测序 分析表明，通过使用前体(Pre)-miR-17恢复人类miR-17水平，对664个基因进行了调节 RASF。核糖核酸测序数据的独创性途径分析确定了肿瘤坏死因子-α中的泛素蛋白酶体系统 信号通路作为miR-17的主要靶点。此外，使用前体-miR-17恢复miR-17水平 17(前miR-17)降低了泛素E3连接酶TRAF2与其信号伙伴cIAP2的结合能力，从而 抑制肿瘤坏死因子-α诱导的下游信号转导通路，抑制IL-6、IL-8的产生 MMP1、MMP13在人RASF中的表达。虽然这些新发现为miR-17的影响提供了证据 RASF中在肿瘤坏死因子-α信号转导中起关键作用的翻译后过程的替换，几个重要的问题 17生物发生和周转仍未得到回答，包括RA严重低表达的原因、影响 促炎症细胞因子对miR-17生物发生和周转的影响以及miR-17的相关性和有效性 类风湿关节炎的替代疗法。基于这些新的观察，我们认为miR-17替代疗法可以 改善类风湿关节炎。因此，在具体目标1中，我们将确定miR-17生物发生和 RA的营业额。在目标2中，我们将评估miR-17修复对肿瘤坏死因子-α的相关性和分子机制。 诱导人RASF的分子和表型变化。最后，Aim 3将测试miR-17的体内疗效 人RA的肿瘤坏死因子-α转基因小鼠模型和大鼠动脉粥样硬化模型。这些研究的成功 将导致两个不同的临床发现：1)阐明改变的miRNA的生物发生和代谢机制 RA的发病机制，以及2)基于miRNA的治疗RA的方法的有效性。