Li Xiaoyu, Sun Hao, Li Hao, Li Deng, Cai Zhiqing, Xu Jie, Ma Ruofan
Department of Orthopaedics, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China.
Department of Anesthesiology, Yunyang County People's Hospital, Chongqing, China.
DNA Cell Biol. 2023 Apr;42(4):212-222. doi: 10.1089/dna.2022.0509. Epub 2023 Mar 20.
The polarization states and molecular signatures of macrophages in the synovium of patients with rheumatoid arthritis (RA) are not well understood. We aimed to identify specific subpopulations of macrophages and their features in RA synovium thereby providing a theoretical basis for treatment of RA. Single-cell RNA sequencing (scRNA-seq) was used to identify cell subsets and their gene signatures in synovial cells of patients with RA and osteoarthritis (OA). Spatial distribution of macrophages was visualized by deconvolving spatial transcriptomic data with scRNA-seq data. Flow cytometry and immunofluorescence were applied to investigate the expression of macrophage polarization indicators CD86 and CD206. Trajectory analysis was used to determine differentiation relationships. Transcription factor (TF) analysis was performed to find specific TFs. scRNA-seq identified three cell clusters of macrophages: M0-like Mϕ1, M2-like Mϕ2, and M1-like Mϕ3. Mϕ1 distributed widely in the synovium, whereas Mϕ2 and Mϕ3 distributed sparsely. CD86 and CD206 were both upregulated in macrophages of RA synovium, especially in lining layer. Trajectory analysis showed that Mϕ1 existed at the start of the differentiation trajectory. , , and were TFs specific to Mϕ1, Mϕ2, and Mϕ3 under RA condition, respectively. Compared with in OA condition, three macrophage clusters upregulated , , , , , , , , , and in NF-kappa B signaling pathway. The identification of macrophage subsets with different polarized states and their molecular signatures provided a more precise understanding of macrophages, which may contribute to the development of novel therapeutic strategy for RA.
类风湿性关节炎（RA）患者滑膜中巨噬细胞的极化状态和分子特征尚未得到充分了解。我们旨在识别RA滑膜中巨噬细胞的特定亚群及其特征，从而为RA的治疗提供理论依据。使用单细胞RNA测序（scRNA-seq）来识别RA和骨关节炎（OA）患者滑膜细胞中的细胞亚群及其基因特征。通过将空间转录组数据与scRNA-seq数据进行反卷积，可视化巨噬细胞的空间分布。应用流式细胞术和免疫荧光来研究巨噬细胞极化指标CD86和CD206的表达。使用轨迹分析来确定分化关系。进行转录因子（TF）分析以找到特定的TF。scRNA-seq识别出巨噬细胞的三个细胞簇：M0样Mϕ1、M2样Mϕ2和M1样Mϕ3。Mϕ1广泛分布于滑膜中，而Mϕ2和Mϕ3分布稀疏。CD86和CD206在RA滑膜巨噬细胞中均上调，尤其是在内衬层。轨迹分析表明Mϕ1存在于分化轨迹的起始阶段。 、 和 分别是RA条件下Mϕ1、Mϕ2和Mϕ3特有的TF。与OA条件相比，三个巨噬细胞簇在NF-κB信号通路中上调了 、 、 、 、 、 、 、 、 和 。识别具有不同极化状态的巨噬细胞亚群及其分子特征，为巨噬细胞提供了更精确的理解，这可能有助于开发RA的新型治疗策略。
