A single-nucleus RNA-sequencing pipeline to decipher the molecular anatomy and pathophysiology of human kidneys
A single-nucleus RNA-sequencing pipeline to decipher the molecular anatomy and pathophysiology of human kidneys
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DOI：
10.1038/s41467-019-10861-2
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发表时间：
2019-06-27
影响因子：
16.6
通讯作者：
Jain, Sanjay
中科院分区：
文献类型：
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作者：
Lake, Blue B.;Chen, Song;Jain, Sanjay
定义肾脏内的细胞和分子身份对于了解其在健康和疾病中的组织和功能是必要的。在这里，我们展示了一种可重复的方法，具有最小的伪影，用于基于单核液滴的RNA测序（snDrop-Seq），我们使用它来解析人类成人肾脏中30个不同的细胞群。我们定义了跨越两个主要肾脏区域的十多个肾元段的分子过渡态。我们进一步描述了与慢性肾脏疾病、糖尿病和高血压相关的基因的细胞类型特异性表达，为可能的靶向治疗提供了见解。这包括在系膜细胞中高血压相关的机械感觉离子通道的表达，以及病原表达特征定义的近端小管细胞群的鉴定。我们完全优化的、质量控制的转录组分析管道构成了一种工具，用于生成适用于临床样品的健康和患病分子图谱。
Defining cellular and molecular identities within the kidney is necessary to understand its organization and function in health and disease. Here we demonstrate a reproducible method with minimal artifacts for single-nucleus Droplet-based RNA sequencing (snDrop-Seq) that we use to resolve thirty distinct cell populations in human adult kidney. We define molecular transition states along more than ten nephron segments spanning two major kidney regions. We further delineate cell type-specific expression of genes associated with chronic kidney disease, diabetes and hypertension, providing insight into possible targeted therapies. This includes expression of a hypertension-associated mechano-sensory ion channel in mesangial cells, and identification of proximal tubule cell populations defined by pathogenic expression signatures. Our fully optimized, quality-controlled transcriptomic profiling pipeline constitutes a tool for the generation of healthy and diseased molecular atlases applicable to clinical samples.