Department of Internal Medicine, Jagiellonian University Medical College, 8 Skawinska Str., 31-066 Kraków, Poland.
Clinic of Rheumatology and Immunology, University Hospital, 2 Jakubowskiego Str., 30-668 Kraków, Poland.
Clin Exp Immunol. 2022 May 13;208(1):103-113. doi: 10.1093/cei/uxac022.
Granulomatosis with polyangiitis is a chronic systemic inflammation of small vessels characterized by circulating anti-proteinase 3 antibodies. MicroRNAs are short transcripts specifically inhibiting protein translation. Neutrophils can release extracellular vesicles (EVs). In this study, we characterized profile of microRNA trafficked by EVs in GPA. Fifty patients with GPA were enrolled in the study, 25 at acute phase and 25 in remission. EVs were isolated from the blood serum, characterized by their number, size distribution. Following unbiased screening for microRNA expression, differentially expressed candidates were measured by quantitative real-time PCR. Circulating DNA-myeloperoxidase complexes and apoptosis-related transcripts in peripheral blood neutrophils were quantified. We identified four differentially expressed microRNAs from EVs in granulomatosis with polyangiitis (GPA). MirRs-223-3p, 664a-3p, and 200b-3p were overexpressed and miR-769-5p suppressed in the disease. A distinction between GPA and healthy controls was the best for miR-223-3p, whereas miR-664a-3p discriminated between active vs. remission of GPA. Correct classification of the disease based on multivariate discriminant analysis was between 92% for acute phase and 85% for all study participants. Bioinformatics tools identified genes transcripts potentially targeted by the microRNAs belonging to pathways of focal adhesion, mTOR signaling and neutrophil extracellular traps formation. Two microRNAs positively correlating with the disease activity were involved in neutrophil extracellular traps formation and apoptosis inhibition. A comprehensive characteristics of microRNAs trafficked in bloodstream inside EVs correlates well with our understanding of the mechanisms of GPA and suggests the importance of EVs in progression of the disease.
肉芽肿性多血管炎是一种小血管的慢性系统性炎症，其特征是循环抗蛋白酶 3 抗体。微小 RNA 是专门抑制蛋白质翻译的短转录本。中性粒细胞可以释放细胞外囊泡 (EVs)。在这项研究中，我们描述了 EVs 转运的 GPA 中的 microRNA 谱。50 例 GPA 患者纳入研究，25 例处于急性期，25 例处于缓解期。从血清中分离 EVs，通过数量、大小分布进行特征分析。对 microRNA 表达进行无偏筛选后，采用定量实时 PCR 测量差异表达的候选物。定量外周血中性粒细胞中循环 DNA-髓过氧化物酶复合物和凋亡相关转录物。我们从肉芽肿性多血管炎 (GPA) 的 EVs 中鉴定出 4 个差异表达的 microRNA。MirRs-223-3p、664a-3p 和 200b-3p 在疾病中过表达，miR-769-5p 受到抑制。miR-223-3p 可区分 GPA 和健康对照，miR-664a-3p 可区分 GPA 活动期与缓解期。基于多变量判别分析，对疾病的正确分类在急性期为 92%，在所有研究参与者中为 85%。生物信息学工具鉴定出潜在受属于粘着斑、mTOR 信号和中性粒细胞胞外陷阱形成途径的 microRNA 靶向的基因转录物。与疾病活动正相关的两个 microRNA 参与中性粒细胞胞外陷阱形成和凋亡抑制。在 EV 内血液中转运的 microRNA 的全面特征与我们对 GPA 发病机制的理解密切相关，并表明 EVs 在疾病进展中的重要性。
