血清microRNA作为强直性脊柱炎早期诊断分子标记物的筛选研究

Ankylosing spondylitis (AS) is an autoimmune disease that primarily affects young and middle-aged people, leading to disability. To date, no specific early diagnostic molecular markers has been identified. In view of the high disability attributable to AS, it is very urgent to explore and establish an early warning model characterized by simplicity, rapidness, high sensitivity and specificity. Recent studies have demonstrated that the abnormal expression patterns of serum miRNAs play a vital role in the occurrence and development of autoimmune diseases. The results of our previous study also showed that the expression patterns of serum miRNA in AS patients were different from the normal control, indicating that serum miRNA could serve as molecular markers for the diagnosis of autoimmune diseases. Based on the results of our previous study, solexa sequencing and quantitative RT-PCR were further performed to analyze large sample size to obtain valuable miRNAs for early diagosis, effect monitoring and the evaluation of prognosis. Additionally, animal model was used to explore the early diagnostic value and molecular mechanism of these miRNAs, in order to provide experimental and theoretical basis for investigating the ideal markers and the new therapeutic targets. The project may fill a gap in the reasearch on the miRNA of AS.

强直性脊柱炎（ankylosing spondylitis, AS）是一种常见于青壮年的自身免疫性疾病，具有极高的致残性。迄今，尚无特异性的早期分子诊断标记物。鉴于其高致残性，当前亟待探索和建立一种简单、快速、敏感性高和特异性强的早期预警模型。近期研究显示，miRNA表达谱的异常在多种自身免疫性疾病发生、发展中起重要作用。我们的前期研究也证实AS患者血清miRNA表达谱与正常人明显不同，提示血清miRNA具有作为分子标记物的潜能。本课题拟在此基础上，进一步运用全基因组Solexa测序扫描和定量PCR验证相结合的手段，通过大样本临床标本检测分析，筛选出在AS早期诊断、疗效监控、预后判断等方面有临床价值的血清miRNA，再利用动物模型研究miRNA的早期诊断价值及分子机制，从而为临床上寻找理想的诊断标记物及新治疗靶点提供实验依据和理论基础，填补国内外在ASmiRNA领域研究的空白。

AS病理过程血清某些miRNA的表达不同于正常人，这些特异性变化的血清miRNA可作为AS新的分子标记物，有助于AS的早期诊断和治疗效果的适时监控，本研究通过Solexa测序技术进行表达谱初筛、TaqMan探针的实时荧光定量PCR技术对初选出的血清miRNA在每份标本上进行逐一验证，对AS中特异变化的miRNA进行了初步研究。一共募集80例AS患者和78例年龄、性别匹配的正常对照，收集他们的血清标本、影像学资料及各量表资料，并探究了两组人群血清miRNA表达差异,分析miRNA表达量和影像学及临床表现之间的相关性。结果表明：(1) AS组病人血清中miR-146a和 miR-155水平显著高于对照组，可作为诊断AS的新的辅助性生物标志物；(2) 后凸畸形大于等于70度的患者血清miR-155水平明显高于后凸小于70度的患者；同时，相比于miR-146a，miR-155表达水平与BASDAI分数、CRP和mSASSS评分有显著正相关，提示miR-155与疾病活动性及胸腰椎后凸程度间存在联系。项目资助发表SCI论文15篇，核心论文6篇，发明专利一项。本项目投入资金70万元，各项支出基本与预算相符，剩余经费计划用于本项目后续支出。