RNA-sequencing from inflamed and normal synovial membrane after anterior cruciate ligament and/or meniscus injuries

Despite ample evidence demonstrating that anterior cruciate ligament (ACL) and meniscus tears are associated with the development of knee osteoarthritis (OA), the contributing factors remain unknown. Synovial inflammation has recently been recognized as a pivotal factor in the pathogenesis of OA. However, there is a lack of data on synovial profiles after ACL or meniscus injuries, which may contribute to posttraumatic OA (PTOA). We performed RNA-seq of 3 inflamed synovial biopsy samples vs 3 normal synovial biopsy samples following ACL and/or meniscus injuries. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and protein–protein interaction (PPI) analyses were performed to investigate mRNAs with significant differences between the inflamed and normal groups.Next, competing endogenous RNA (ceRNA) networks were constructed based on bioinformatics analyses and quantitative real-time polymerase chain reaction (RT–PCR) results. The association of the identified DEGs with the levels of infiltrating immune cells was explored using Pearson correlation analysis in R software. We aimed to provide greater insights into molecular mechanisms and biologic pathways in synovitis that could regulate post-trauma osteoarthritis progression. Comparing lncRNAs, miRNA and mRNA expression profiling from inflamed and normal synovial tisssue following anterior cruciate ligament and/or meniscus injuries

尽管已有大量研究证据表明，前交叉韧带（anterior cruciate ligament, ACL）损伤与半月板撕裂（meniscus tears）均与膝骨关节炎（knee osteoarthritis, OA）的发生发展密切相关，但其具体致病贡献因素仍未明确。近年来，滑膜炎症（synovial inflammation）已被证实为骨关节炎发病机制（pathogenesis）中的关键核心因素。然而，目前针对前交叉韧带或半月板损伤后滑膜分子特征的相关研究数据仍较为匮乏，而这一环节可能与创伤后骨关节炎（posttraumatic OA, PTOA）的发生存在紧密关联。本研究对前交叉韧带和/或半月板损伤后获取的3份炎性滑膜活检样本（synovial biopsy samples）与3份正常滑膜活检样本开展了RNA测序（RNA-seq）分析。通过基因本体论（Gene Ontology, GO）富集分析、京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路分析以及蛋白质-蛋白质相互作用（protein–protein interaction, PPI）网络分析，系统探究了炎性与正常滑膜组间存在显著表达差异的mRNA分子。后续，基于生物信息学分析结果与实时定量聚合酶链式反应（quantitative real-time polymerase chain reaction, RT–PCR）验证数据，构建了内源竞争RNA（competing endogenous RNA, ceRNA）调控网络。借助R软件（R software）中的Pearson相关分析（Pearson correlation analysis），本研究进一步探究了所鉴定的差异表达基因（differentially expressed genes, DEGs）与滑膜组织浸润免疫细胞水平之间的潜在关联。本研究旨在深入解析滑膜炎症中调控创伤后骨关节炎进展的分子机制与生物学通路，并对比分析前交叉韧带和/或半月板损伤后炎性与正常滑膜组织的长链非编码RNA（long non-coding RNA, lncRNA）、微小RNA（microRNA, miRNA）及mRNA表达谱特征。