目的:评估滑液(SF)中microRNA的表达,等离子体,和幼年特发性关节炎(JIA)患者的白细胞。
方法:通过绝对定量液滴数字PCR阵列评估合并的JIA血浆和SF中的MicroRNA表达。在个体患者样品中验证结果。通过实时PCR在JIA血液和SF中评估白细胞和细胞外囊泡中的MicroRNA含量。将血液microRNA表达与健康对照(HC)进行比较。主成分分析用于描述JIA血浆和SFmicroRNAs,并通过途径分析研究了microRNA失调的潜在生物学后果。
结果:JIA血浆中的MiR-15a-5p和miR-409-3p水平高于HC血浆。JIASF含有升高水平的miR-21-5p,miR-27a-3p,miR-146b-5p,miR-155-5p,和miR-423-5p,并降低miR-192-5p和miR-451a,与JIA等离子体相比。胞外囊泡分析显示选定的microRNA之间的可变包封,只有miR-155-5p在细胞外囊泡中基本上存在。SF白细胞也有较高的miR-21-5p表达,miR-27a-3p,miR-146b-5p,还有miR-155-5p,miR-409-3p和miR-451a的表达降低,相对于血液。在JIA和HC血液白细胞之间没有观察到差异。与JIA血液样品相比,JIA关节液和白细胞中的microRNA簇通常会发生改变。计算机模拟分析预测JIA中差异表达的microRNAs靶向转化生长因子(TGF)-β途径。
结论:在JIA中,多种microRNA的表达在局部和系统上都失调,这可能会抑制TGF-β途径。这些发现提高了我们对JIA免疫发病机制的认识,并可能导致靶向治疗的发展。
To evaluate microRNA expression in synovial fluid (SF), plasma, and leukocytes from patients with juvenile idiopathic arthritis (JIA).
MicroRNA expression in pooled JIA plasma and SF was assessed by absolute quantitative droplet digital PCR array. The results were validated in individual patient samples. MicroRNA content in leukocytes and extracellular vesicles was evaluated by real-time PCR in JIA blood and SF. Blood microRNA expression was compared with healthy controls (HCs). Principal component analysis was used to profile JIA plasma and SF microRNAs, and the potential biological consequences of microRNA dysregulation were investigated by pathway analysis.
MiR-15a-5p and miR-409-3p levels were higher in JIA plasma than in HC plasma. JIA SF contained elevated levels of miR-21-5p, miR-27a-3p, miR-146b-5p, miR-155-5p, and miR-423-5p, and decreased miR-192-5p and miR-451a, compared to JIA plasma. Extracellular vesicle analysis demonstrated variable encapsulation among selected microRNAs, with only miR-155-5p being represented substantially in extracellular vesicles. SF leukocytes also had higher expression of miR-21-5p, miR-27a-3p, miR-146b-5p, and miR-155-5p, and lower expression of miR-409-3p and miR-451a, relative to blood. No differences were observed between JIA and HC blood leukocytes. Clusters of microRNAs were commonly altered in JIA joint fluid and leukocytes compared to JIA blood samples. In silico analysis predicted that differentially expressed microRNAs in JIA target the transforming growth factor (TGF)-β pathway.
The expression of multiple microRNAs is dysregulated in JIA both locally and systemically, which may inhibit the TGF-β pathway. These findings advance our knowledge of JIA immunopathogenesis and may lead to the development of targeted therapies.