Jiang Jianjun, Liu Yang
Department of General Surgery, Vascular Surgery, Qilu Hospital of Shandong University, Jinan, Shandong, People's Republic of China.
J Inflamm Res. 2023 Jun 8;16:2431-2436. doi: 10.2147/JIR.S405903. eCollection 2023.
A comprehensive study of the distribution and role of immune/inflammatory cells in thrombosis is still lacking because traditional pathology techniques cannot accomplish the analysis of numerous protein and genetic data simultaneously. We aimed to evaluate the feasibility of digital spatial profiling (DSP) to study immune/inflammation reaction in thrombosis progression.
An 82-year-old male patient underwent iliofemoral thrombectomy at our institution. The white, mixed and red thrombi were fixed in formalin, dehydrated in ethanol and embedded in paraffin, which were incubated with morphology-labeled fluorescent antibodies (CD45, SYTO13) and the entire target mixture in GeoMx Whole Transcriptome Atlas panel. DSP system was applied to investigate the regions of interest from fluorescence imaging. Fluorescence imaging showed infiltration of immune/inflammation cells in white, mixed and red thrombosis. Whole genome sequencing revealed 16 genes differentially expressed. Pathway enrichment analysis revealed that these genes were significantly enriched in ligand binding and uptake related signaling pathways of the scavenger receptor. The distribution of immune/inflammation cell subsets was different in white, mixed and red thrombosis. The abundance of endothelial cells, CD8 naive T cells, and macrophages in red thrombosis was significantly higher than in mixed and white thrombosis.
The results showed that DSP can facilitate efficient analysis using very few thrombosis samples and provide valuable new leads, suggesting that DSP may be a viable and important new tool to study thrombosis and inflammation.
由于传统病理学技术无法同时分析大量蛋白质和基因数据，因此仍缺乏对血栓形成过程中免疫/炎症细胞分布和作用的全面研究。我们旨在评估数字空间分析（DSP）在研究血栓形成过程中免疫/炎症反应的可行性。
一名82岁男性患者在我院接受了髂股血栓切除术。白色、混合性和红色血栓用福尔马林固定，乙醇脱水，石蜡包埋，然后与形态学标记的荧光抗体（CD45、SYTO13）以及GeoMx全转录组图谱面板中的整个靶标混合物一起孵育。应用DSP系统从荧光成像中研究感兴趣的区域。荧光成像显示白色、混合性和红色血栓中有免疫/炎症细胞浸润。全基因组测序揭示了16个差异表达基因。通路富集分析表明，这些基因在清道夫受体的配体结合和摄取相关信号通路中显著富集。白色、混合性和红色血栓中免疫/炎症细胞亚群的分布不同。红色血栓中内皮细胞、CD8幼稚T细胞和巨噬细胞的丰度显著高于混合性和白色血栓。
结果表明，DSP可以用极少的血栓样本促进高效分析，并提供有价值的新线索，提示DSP可能是研究血栓形成和炎症的一种可行且重要的新工具。
