Single cell and spatial transcriptomics analysis of kidney double negative T lymphocytes in normal and ischemic mouse kidneys.

Single cell and spatial transcriptomics analysis of kidney double negative T lymphocytes in normal and ischemic mouse kidneys.
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DOI:
10.1038/s41598-023-48213-2
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发表时间:
2023-11-28
期刊:
影响因子:
4.6
通讯作者:
--
中科院分区:
综合性期刊3区
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--
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T细胞在急性肾损伤(AKI)的发病机制中很重要,TCR+CD4-CD8-(双阴性-DN)是具有调节特性的T细胞。然而,与传统的 CD4+ 和 CD8+ 细胞相比,有关 DN T 细胞的信息有限。为了阐明 AKI 期间 DN T 细胞的分子特征和空间动力学,我们对分选的小鼠 DN、CD4+ 和 CD8+ 细胞进行单细胞 RNA 测序 (scRNA-seq),并结合正常和 AKI 后小鼠肾脏的空间转录组分析。 scRNA-seq 揭示了小鼠肾脏 DN、CD4+ 和 CD8+ T 细胞的不同转录谱,与正常肾组织中的 CD4+ 和 CD8+ T 细胞相比,DN T 细胞中 Kcnq5、Klrb1c、Fcer1g 和 Klre1 表达富集。我们利用 RT-PCR 以及 NIH 人类肾脏精准医学项目 (KPMP) 的 Kcnq5、Klrb1 和 Fcer1g 基因验证了这四种基因在小鼠肾脏 DN、CD4+ 和 CD8+ T 细胞中的表达。正常和缺血小鼠肾组织的空间转录组学显示,外髓中有局部 T 细胞簇,表达包括 Fcer1g 在内的 DN T 细胞基因。这些结果为未来研究正常和患病肾脏中的 DN T 以及 CD4+ 和 CD8+ 细胞提供了模板。
T cells are important in the pathogenesis of acute kidney injury (AKI), and TCR+CD4-CD8- (double negative-DN) are T cells that have regulatory properties. However, there is limited information on DN T cells compared to traditional CD4+ and CD8+ cells. To elucidate the molecular signature and spatial dynamics of DN T cells during AKI, we performed single-cell RNA sequencing (scRNA-seq) on sorted murine DN, CD4+, and CD8+ cells combined with spatial transcriptomic profiling of normal and post AKI mouse kidneys. scRNA-seq revealed distinct transcriptional profiles for DN, CD4+, and CD8+ T cells of mouse kidneys with enrichment of Kcnq5, Klrb1c, Fcer1g, and Klre1 expression in DN T cells compared to CD4+ and CD8+ T cells in normal kidney tissue. We validated the expression of these four genes in mouse kidney DN, CD4+ and CD8+ T cells using RT-PCR and Kcnq5, Klrb1, and Fcer1g genes with the NIH human kidney precision medicine project (KPMP). Spatial transcriptomics in normal and ischemic mouse kidney tissue showed a localized cluster of T cells in the outer medulla expressing DN T cell genes including Fcer1g. These results provide a template for future studies in DN T as well as CD4+ and CD8+ cells in normal and diseased kidneys.
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