Single-Cell RNA Profiling of Glomerular Cells Shows Dynamic Changes in Experimental Diabetic Kidney Disease
Single-Cell RNA Profiling of Glomerular Cells Shows Dynamic Changes in Experimental Diabetic Kidney Disease
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DOI：
10.1681/asn.2018090896
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发表时间：
2019-04-01
影响因子：
13.6
通讯作者：
He, John Cijiang
中科院分区：
文献类型：
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作者：
Fu, Jia;Akat, Kemal M.;He, John Cijiang
最近的单细胞RNA测序（scRNA-seq）分析提供了对正常肾脏细胞特异性基因表达谱的深入了解。然而，在病变肾脏中，对特定细胞，特别是肾小球细胞的变化的理解仍然有限。方法为了阐明糖尿病肾病肾小球细胞特异性基因表达的变化，我们对来自链脲佐菌素诱导的糖尿病内皮型一氧化氮合酶（eNOS）缺陷的肾小球细胞进行了scRNA-seq分析。结果我们鉴定了5种不同的细胞群，包括肾小球内皮细胞、系膜细胞、足细胞、免疫细胞和肾小管细胞。使用scRNA-seq分析，我们证实了肾小球细胞特异性标志物的表达，并鉴定了几种新的肾小球细胞潜在标志物。糖尿病肾小球中免疫细胞的数量显著高于对照肾小球，进一步的聚类分析显示这些免疫细胞主要是巨噬细胞。对糖尿病小鼠和对照小鼠的内皮细胞和系膜细胞中差异基因表达的分析表明，表达基因的模式发生动态变化，其中许多基因已知与糖尿病肾病有关。此外，基因表达分析表明，个别细胞的糖尿病injuries.Conclusions变量的反应，我们的研究结果表明，scRNA-seq分析的能力，在分离的肾小球细胞从糖尿病和对照小鼠，揭示糖尿病肾脏基因表达的动态变化，与变量的个别细胞的反应。这种变化，这可能是不明显的，在大量的肾小球细胞的转录组学分析，可能有助于确定重要的病理生理因素，有助于糖尿病肾病的进展。
Background Recent single-cell RNA sequencing (scRNA-seq) analyses have offered much insight into cell-specific gene expression profiles in normal kidneys. However, in diseased kidneys, understanding of changes in specific cells, particularly glomerular cells, remains limited.Methods To elucidate the glomerular cell-specific gene expression changes in diabetic kidney disease, we performed scRNA-seq analysis of isolated glomerular cells from streptozotocin-induced diabetic endothelial nitric oxide synthase (eNOS)-deficient (eNOS(-/-)) mice and control eNOS(-/-) mice.Results We identified five distinct cell populations, including glomerular endothelial cells, mesangial cells, podocytes, immune cells, and tubular cells. Using scRNA-seq analysis, we confirmed the expression of glomerular cell-specific markers and also identified several new potential markers of glomerular cells. The number of immune cells was significantly higher in diabetic glomeruli compared with control glomeruli, and further cluster analysis showed that these immune cells were predominantly macrophages. Analysis of differential gene expression in endothelial and mesangial cells of diabetic and control mice showed dynamic changes in the pattern of expressed genes, many of which are known to be involved in diabetic kidney disease. Moreover, gene expression analysis showed variable responses of individual cells to diabetic injury.Conclusions Our findings demonstrate the ability of scRNA-seq analysis in isolated glomerular cells from diabetic and control mice to reveal dynamic changes in gene expression in diabetic kidneys, with variable responses of individual cells. Such changes, which might not be apparent in bulk transcriptomic analysis of glomerular cells, may help identify important pathophysiologic factors contributing to the progression of diabetic kidney disease.