背景N-糖基化是人类中最常见的翻译后修饰之一。这些改变与肾脏疾病相关。方法一种新颖的技术方法，单细胞N-乙酰乳糖胺测序（scLacNAc-seq），应用于同时检测N-糖基化表达和转录组的单细胞分辨率在三个人肾组织从零次活检。细胞簇,每个细胞簇中的糖化丰度，功能富集分析，小区-小区串扰，并应用伪时间分析。结果使用scLacNAc-seq，确定了24,247个细胞和22个细胞簇，并且获得每个细胞中的N-聚糖丰度。转录组分析显示毛细血管内皮细胞（CapEC）和壁上皮细胞（PEC）之间存在密切联系。PEC和CapEC通过几对配体受体相互通信(例如，TGFB1-EGFR,GRN-EGFR,TIMP1-FGFR2,VEGFB-FLT1,ANGPT2-TEK,和GRN-TNFRSF1A)。最后,构建了PEC和CapEC之间的小区-小区串扰调节网络，参与细胞发育。结论我们在这里，第一次,从零时间活检中构建了人肾脏中22个细胞簇的糖基化谱。此外，PEC和CapEC之间通过配体-受体系统的细胞-细胞通讯可能在细胞增殖中起着至关重要的调节作用。
BACKGROUND: N-glycosylation is one of the most common posttranslational modifications in humans, and these alterations are associated with kidney diseases.
METHODS: A novel technological approach, single-cell N-acetyllactosamine sequencing (scLacNAc-seq), was applied to simultaneously detect N-glycosylation expression and the transcriptome at single-cell resolution in three human kidney tissues from zero-time biopsy. Cell clusters, glycation abundance in each cell cluster, functional enrichment analysis, cell-cell crosstalk, and pseudotime analysis were applied.
RESULTS: Using scLacNAc-seq, 24,247 cells and 22 cell clusters were identified, and N-glycan abundance in each cell was obtained. Transcriptome analysis revealed a close connection between capillary endothelial cells (CapECs) and parietal epithelial cells (PECs). PECs and CapECs communicate with each other through several pairs of ligand receptors (e.g., TGFB1-EGFR, GRN-EGFR, TIMP1-FGFR2, VEGFB-FLT1, ANGPT2-TEK, and GRN-TNFRSF1A). Finally, a regulatory network of cell-cell crosstalk between PECs and CapECs was constructed, which is involved in cell development.
CONCLUSIONS: We here, for the first time, constructed the glycosylation profile of 22 cell clusters in the human kidney from zero-time biopsy. Moreover, cell-cell communication between PECs and CapECs through the ligand-receptor system may play a crucial regulatory role in cell proliferation.
