Queen Mary University of London, London, United Kingdom.
Queen Mary University of London and Kings College London, Guys' Campus, London, United Kingdom.
Arthritis Rheumatol. 2024 Dec;76(12):1705-1718. doi: 10.1002/art.42958. Epub 2024 Aug 16.
This study was undertaken to establish the potential therapeutic profile of neutrophil-derived extracellular vesicles (EVs) in experimental inflammatory arthritis and associate pharmacological activity with specific EV components, focusing on microRNAs.
Neutrophil EVs were administered intra-articularly through a prophylactic or therapeutic protocol to male C57BL/6 mice undergoing serum-transfer-induced inflammatory arthritis. Transcriptomic analysis of knees was performed on joints following EV administration, naive and arthritic mice (untreated; n = 4/group) and EV-treated diseased mice (intra-articular administration) with contralateral (vehicle-treated; n = 8/group). Comparison of healthy donor and patients with rheumatoid arthritis (RA) neutrophil EVs was performed.
EVs afforded cartilage protection with an increase in collagen-II and reduced collagen-X expression within the joint. To gain mechanistic insights, RNA sequencing of the arthritic joints was conducted. A total of 5,231 genes were differentially expressed (P < 0.05), with 257 unique to EV treatment. EVs affected key regenerative pathways involved in joint development, including Wnt and Notch signaling. This wealth of genomic alteration prompted to identify microRNAs in EVs, 10 of which are associated with RA. As a proof of concept, we focused on miR-455-3p, which was detected in both healthy donor and RA EVs. EV addition to chondrocyte cultures elevated miR-455-3p and exerted anticatabolic effects upon interleukin-1β stimulation; these effects were blocked by actinomycin or miR-455-3p antagomir.
Neutrophils from patients with RA yielded EVs with composition, efficacy, and miR-455-3p content similar to those of healthy volunteers, suggesting that neutrophil EVs could be developed as an autologous treatment to protect and repair joint tissue of patients affected by inflammatory arthritides.
本研究旨在确定中性粒细胞衍生的细胞外囊泡（EVs）在实验性炎症性关节炎中的潜在治疗作用，并将药理活性与特定的 EV 成分（重点是 microRNAs）相关联。
通过预防性或治疗性方案向接受血清转移诱导的炎症性关节炎的雄性 C57BL/6 小鼠关节内给予中性粒细胞 EV。在 EV 给药后对膝关节进行转录组分析，包括未处理的关节（naive）和关节炎小鼠（未治疗；每组 n=4）以及接受 EV 治疗的患病关节（关节内给药；每组 n=8）。比较健康供体和类风湿关节炎（RA）患者中性粒细胞 EVs。
EV 提供了软骨保护作用，增加了关节内的胶原-II 表达，降低了胶原-X 的表达。为了获得机制见解，对关节炎关节进行了 RNA 测序。总共检测到 5231 个差异表达的基因（P<0.05），其中 257 个基因是 EV 治疗所特有的。EV 影响了参与关节发育的关键再生途径，包括 Wnt 和 Notch 信号通路。这种丰富的基因组改变促使我们在 EV 中鉴定 microRNAs，其中 10 个与 RA 相关。作为概念验证，我们专注于 miR-455-3p，它在健康供体和 RA EV 中均被检测到。在软骨细胞培养物中添加 EV 会增加 miR-455-3p 的表达，并在白细胞介素-1β刺激下发挥抗分解代谢作用；这些作用被放线菌素或 miR-455-3p 反义寡核苷酸阻断。
来自 RA 患者的中性粒细胞产生的 EV 具有与健康志愿者相似的组成、功效和 miR-455-3p 含量，这表明中性粒细胞 EV 可以作为一种自体治疗方法，用于保护和修复受炎症性关节炎影响的患者的关节组织。
