Altered glucose metabolism in proximal tubule cells responding to acute kidney injury is associated with patient mortality

Single-cell transcriptomics revealed that gluconeogenesis is impaired in proximal tubule cells during AKI in mice. To investigate single cell transcriptomics on renal tubule cells, we generated a reporter mouse expressing GFP linked to the histone protein H2B in all nephron cells by combining 3 transgenes Six2-TGC; Rosa26rtTA ; pTRE-H2BeGFP. IRI and Sham surgeries were performed, and kidney tissue was harvested at different timepoints, followed by nuclei isolation, FACs sorting, and 10X Genomics Chromium Drop-Seq and library construction. Illumina Next-Seq sequencing of barcoded single nuclei libraries was performed and aligned to a pre-mrna genome with STAR/CellRanger. Gene/barcode matrices were merged and processed with the Seurat R package.

单细胞转录组学（single-cell transcriptomics）研究表明，小鼠急性肾损伤（AKI）进程中，近端小管细胞的糖异生功能受损。为探究肾小管细胞的单细胞转录组学特征，我们通过整合3种转基因元件（Six2-TGC、Rosa26rtTA、pTRE-H2BeGFP），构建了一株在所有肾单位细胞中表达与组蛋白H2B融合的绿色荧光蛋白（GFP）的报告基因小鼠。随后开展缺血再灌注（IRI）与假手术（Sham）造模，于不同时间点收取肾脏组织，依次进行细胞核分离、荧光激活细胞分选（FACs）、10X Genomics Chromium Drop-Seq建库。采用Illumina Next-Seq平台对带条形码的单细胞核文库进行测序，并通过STAR/CellRanger软件将测序数据比对至pre-mRNA基因组。最终通过Seurat R包对基因/条形码矩阵进行合并与分析处理。