Colombini Alessandra, Libonati Francesca, Lopa Silvia, Ragni Enrico, De Luca Paola, Zagra Luigi, Sinigaglia Federico, Moretti Matteo, de Girolamo Laura
Laboratorio di Biotecnologie Applicate all'Ortopedia, IRCCS Istituto Ortopedico Galeazzi, Milan, Italy.
Cell and Tissue Engineering Laboratory, IRCCS Istituto Ortopedico Galeazzi, Milan, Italy.
Front Med (Lausanne). 2022 Oct 12;9:992386. doi: 10.3389/fmed.2022.992386. eCollection 2022.
The purpose of the present study is to predict by bioinformatics the activity of the extracellular vesicle (EV)-embedded micro RNA (miRNAs) secreted by cartilage cells (CCs), adipose tissue-derived- (ASCs), and bone marrow-derived stem cells (BMSCs) and verify their immunomodulatory potential supporting our bioinformatics findings to optimize the autologous cell-based therapeutic strategies for osteoarthritis (OA) management. Cells were isolated from surgical waste tissues of three patients who underwent total hip replacement, expanded and the EVs were collected. The expression of EV-embedded miRNA was evaluated with the QuantStudio 12 K Flex OpenArray platform. Mientournet and ingenuity pathway analysis (IPA) were used for validated target prediction analysis and to identify miRNAs involved in OA and inflammation. Cells shared the expression of 325 miRNAs embedded in EVs and differed for the expression of a small number of them. Mienturnet revealed no results for miRNAs selectively expressed by ASCs, whereas miRNA expressed by CCs and BMSCs were putatively involved in the modulation of cell cycle, senescence, apoptosis, Wingless and Int-1 (Wnt), transforming growth factor beta (TGFβ), vascular endothelial growth factor (VEGF), Notch, Hippo, tumor necrosis factor alpha (TNFα), interleukin 1 beta (IL-1β), insulin like growth factor 1 (IGF-1), RUNX family transcription factor 2 (RUNX2), and endochondral ossification pathways. Cartilage homeostasis, macrophages and T cells activity and inflammatory mediators were identified by IPA as targets of the miRNAs found in all the cell populations. Co-culture tests on macrophages and T cells confirmed the immuno-modulatory ability of CCs, ASCs, and BMSCs. The study findings support the rationale behind the use of cell-based therapy for the treatment of OA.
本研究的目的是通过生物信息学预测软骨细胞（CCs）、脂肪组织来源的干细胞（ASCs）和骨髓来源的干细胞（BMSCs）分泌的细胞外囊泡（EV）包裹的微小RNA（miRNAs）的活性，并验证其免疫调节潜力，以支持我们的生物信息学研究结果，从而优化用于骨关节炎（OA）管理的基于自体细胞的治疗策略。从三名接受全髋关节置换术的患者的手术废弃物组织中分离细胞，进行扩增并收集EV。使用QuantStudio 12 K Flex OpenArray平台评估EV包裹的miRNA的表达。使用Mientournet和 Ingenuity通路分析（IPA）进行验证的靶标预测分析，并确定参与OA和炎症的miRNAs。细胞共享325种EV包裹的miRNAs的表达，其中少数miRNAs的表达存在差异。Mienturnet显示未找到ASCs选择性表达的miRNAs的结果，而CCs和BMSCs表达的miRNAs可能参与细胞周期、衰老、凋亡、无翅型MMTV整合位点家族（Wnt）、转化生长因子β（TGFβ）、血管内皮生长因子（VEGF）、Notch、Hippo、肿瘤坏死因子α（TNFα）、白细胞介素1β（IL-1β）、胰岛素样生长因子1（IGF-1）、RUNX家族转录因子2（RUNX2）和软骨内成骨途径的调节。通过IPA确定软骨稳态、巨噬细胞和T细胞活性以及炎症介质是所有细胞群体中发现的miRNAs的靶标。对巨噬细胞和T细胞的共培养试验证实了CCs、ASCs和BMSCs的免疫调节能力。研究结果支持了使用基于细胞的疗法治疗OA的基本原理。
