Department of Gene Technology, Royal Institute of Technology, Science for Life Laboratory, Stockholm, Sweden.
Division of Rheumatology, Department of Medicine Solna, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden.
Sci Rep. 2019 Dec 12;9(1):18975. doi: 10.1038/s41598-019-55441-y.
Lately it has become possible to analyze transcriptomic profiles in tissue sections with retained cellular context. We aimed to explore synovial biopsies from rheumatoid arthritis (RA) and spondyloarthritis (SpA) patients, using Spatial Transcriptomics (ST) as a proof of principle approach for unbiased mRNA studies at the site of inflammation in these chronic inflammatory diseases. Synovial tissue biopsies from affected joints were studied with ST. The transcriptome data was subjected to differential gene expression analysis (DEA), pathway analysis, immune cell type identification using Xcell analysis and validation with immunohistochemistry (IHC). The ST technology allows selective analyses on areas of interest, thus we analyzed morphologically distinct areas of mononuclear cell infiltrates. The top differentially expressed genes revealed an adaptive immune response profile and T-B cell interactions in RA, while in SpA, the profiles implicate functions associated with tissue repair. With spatially resolved gene expression data, overlaid on high-resolution histological images, we digitally portrayed pre-selected cell types in silico. The RA displayed an overrepresentation of central memory T cells, while in SpA effector memory T cells were most prominent. Consequently, ST allows for deeper understanding of cellular mechanisms and diversity in tissues from chronic inflammatory diseases.
最近，人们已经可以在保留细胞结构的组织切片中分析转录组谱。我们旨在通过空间转录组学（ST）探索类风湿关节炎（RA）和脊柱关节炎（SpA）患者的滑膜活检，将其作为在这些慢性炎症性疾病炎症部位进行无偏mRNA 研究的原理验证方法。我们用 ST 研究了受累关节的滑膜组织活检。对转录组数据进行差异基因表达分析（DEA）、途径分析、Xcell 分析鉴定免疫细胞类型，并通过免疫组织化学（IHC）进行验证。ST 技术允许对感兴趣的区域进行选择性分析，因此我们分析了单核细胞浸润的形态学不同区域。最显著的差异表达基因揭示了 RA 中的适应性免疫反应特征和 T-B 细胞相互作用，而在 SpA 中，这些特征涉及与组织修复相关的功能。利用空间分辨的基因表达数据，并与高分辨率组织学图像叠加，我们在数字上描绘了预先选择的细胞类型。RA 中中枢记忆 T 细胞过度表达，而 SpA 中效应记忆 T 细胞最为突出。因此，ST 允许更深入地了解慢性炎症性疾病组织中的细胞机制和多样性。
