Single-Cell RNA Sequencing Reveals Renal Endothelium Heterogeneity and Metabolic Adaptation to Water Deprivation
Single-Cell RNA Sequencing Reveals Renal Endothelium Heterogeneity and Metabolic Adaptation to Water Deprivation
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DOI：
10.1681/asn.2019080832
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发表时间：
2020-01-01
影响因子：
13.6
通讯作者：
Carmeliet, Peter
中科院分区：
文献类型：
--
作者：
Dumas, Sebastien J.;Meta, Elda;Carmeliet, Peter
来自肾小球、皮质和髓质的肾内皮细胞暴露于不同的微环境条件下，并支持特定的肾脏过程。然而，这些细胞的异质性表型仍然不完全清点。渗透稳态对于调节细胞体积和功能至关重要，并且在哺乳动物中，渗透平衡通过肾髓质中的逆流系统调节，其中通过内皮的水交换逆着渗透压梯度发生。脱水暴露髓质肾内皮细胞极端高渗，以及这些细胞如何适应和生存在这种高渗environments.Methods我们编目肾内皮细胞的异质性通过单细胞RNA测序> 40，000小鼠肾内皮细胞，并研究转录组的变化，在渗透适应缺水。我们验证了我们的研究结果，通过免疫染色和功能，通过靶向氧化磷酸化在体外高渗模型和脱水的小鼠在vivo.Results我们确定了24肾内皮细胞表型（其中8个是新的），突出了广泛的异质性，这些细胞之间和内皮质，肾小球和髓质。在响应脱水和高渗，肾髓质内皮细胞上调参与缺氧反应，糖酵解，和令人惊讶的氧化磷酸化的基因的表达。当暴露于高渗时，内皮细胞增加耗氧量，而阻断氧化磷酸化在高渗应激期间损害内皮细胞活力，并在脱水期间损害尿浓度。结论这项研究提供了肾内皮的高分辨率图谱，并突出了广泛的肾内皮细胞表型异质性，以及先前未认识到的氧化磷酸化在肾髓质内皮细胞对缺水的代谢适应中的作用。
Background Renal endothelial cells from glomerular, cortical, and medullary kidney compartments are exposed to different microenvironmental conditions and support specific kidney processes. However, the heterogeneous phenotypes of these cells remain incompletely inventoried. Osmotic homeostasis is vitally important for regulating cell volume and function, and in mammals, osmotic equilibrium is regulated through the countercurrent system in the renal medulla, where water exchange through endothelium occurs against an osmotic pressure gradient. Dehydration exposes medullary renal endothelial cells to extreme hyperosmolarity, and how these cells adapt to and survive in this hypertonic milieu is unknown.Methods We inventoried renal endothelial cell heterogeneity by single-cell RNA sequencing >40,000 mouse renal endothelial cells, and studied transcriptome changes during osmotic adaptation upon water deprivation. We validated our findings by immunostaining and functionally by targeting oxidative phosphorylation in a hyperosmolarity model in vitro and in dehydrated mice in vivo.Results We identified 24 renal endothelial cell phenotypes (of which eight were novel), highlighting extensive heterogeneity of these cells between and within the cortex, glomeruli, and medulla. In response to dehydration and hypertonicity, medullary renal endothelial cells upregulated the expression of genes involved in the hypoxia response, glycolysis, and-surprisingly-oxidative phosphorylation. Endothelial cells increased oxygen consumption when exposed to hyperosmolarity, whereas blocking oxidative phosphorylation compromised endothelial cell viability during hyperosmotic stress and impaired urine concentration during dehydration.Conclusions This study provides a high-resolution atlas of the renal endothelium and highlights extensive renal endothelial cell phenotypic heterogeneity, as well as a previously unrecognized role of oxidative phosphorylation in the metabolic adaptation of medullary renal endothelial cells to water deprivation.