A Single-Cell RNA-Sequencing Analysis of Distinct Subsets of Synovial Macrophages in Rheumatoid Arthritis.
A Single-Cell RNA-Sequencing Analysis of Distinct Subsets of Synovial Macrophages in Rheumatoid Arthritis.
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DOI：
10.1089/dna.2022.0509
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发表时间：
2023-03
影响因子：
3.1
通讯作者：
Xiao-Ye Li;Hao Sun;Hao Li;Deng Li;Zhiqing Cai;Jie Xu;Ruofan Ma
中科院分区：
文献类型：
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作者：
Xiao-Ye Li;Hao Sun;Hao Li;Deng Li;Zhiqing Cai;Jie Xu;Ruofan Ma
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不清楚。本研究旨在明确RA滑膜中巨噬细胞的亚群及其特征，为RA的治疗提供理论依据。使用单细胞RNA测序（scRNA-seq）来鉴定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数据去卷积来可视化巨噬细胞的空间分布。流式细胞术和免疫荧光法检测巨噬细胞极化指标CD 86和CD 206的表达。轨迹分析用于确定分化关系。进行转录因子（TF）分析以找到特异性TF。scRNA-seq鉴定了三个巨噬细胞细胞簇：M0样MARCO+ M11、M2样CSF 1 R + M12和M1样PLAUR+ M13。M β 1广泛分布于滑膜，M β 2和M β 3分布稀疏。CD 86和CD 206在RA滑膜巨噬细胞中均表达上调，以衬里层表达最强。轨迹分析表明，M β 1存在于分化轨迹的开始。HOXB 6、STAT 1和NF KB 2分别是RA条件下对M β 1、M β 2和M β 3特异的转录因子。与OA组相比，3个巨噬细胞团上调NF-κ B B信号通路中的CXCL 2、CXCL 1、IL 1 B、TNFAIP 3、ICAM 1、CXCL 3、PLAU、CCL 4 L2、CCL 4和TNF。不同极化状态的巨噬细胞亚群及其分子特征的鉴定提供了对巨噬细胞的更精确的理解，这可能有助于开发新的RA治疗策略。
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 MARCO+ Mϕ1, M2-like CSF1R+ Mϕ2, and M1-like PLAUR+ 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. HOXB6, STAT1, and NFKB2 were TFs specific to Mϕ1, Mϕ2, and Mϕ3 under RA condition, respectively. Compared with in OA condition, three macrophage clusters upregulated CXCL2, CXCL1, IL1B, TNFAIP3, ICAM1, CXCL3, PLAU, CCL4L2, CCL4, and TNF 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.