Comparison of Human and Murine Enteroendocrine Cells by Transcriptomic and Peptidomic Profiling
Comparison of Human and Murine Enteroendocrine Cells by Transcriptomic and Peptidomic Profiling
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DOI：
10.2337/db18-0883
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发表时间：
2019-05-01
期刊：
影响因子：
7.7
通讯作者：
Gribble, Fiona M.
中科院分区：
文献类型：
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作者：
Roberts, Geoffrey P.;Larraufie, Pierre;Gribble, Fiona M.
肠内分泌细胞 (EEC) 产生胰高血糖素样肽 1 和肽 YY 等激素，调节食物吸收、胰岛素分泌和食欲。基于胰高血糖素样肽 1 疗法治疗 2 型糖尿病和肥胖症的成功，EEC 本身就是增强肠道激素分泌的药物发现计划的焦点。本研究的目的是鉴定人类 EEC 的转录组和肽组，并提供人类和小鼠之间的跨物种比较。通过对细胞固定和染色后通过流式细胞术纯化的人类 EEC 进行 RNA 测序，我们对人类 EEC 群体进行了首次转录组分析，并证明与小鼠对应物具有很强的相关性。 RNA测序的深度足以识别低丰度转录本，例如G蛋白偶联受体和离子通道，揭示了先前发现在餐后营养检测中发挥作用的G蛋白偶联受体在人类EEC中的表达。通过液相色谱-串联质谱法，我们分析了人类和小鼠肠道中肽激素的梯度，包括它们的序列和翻译后修饰。人类和小鼠 EEC 的转录组和肽组谱以及跨物种比较将成为药物发现计划以及了解人类新陈代谢和减肥手术内分泌影响的宝贵工具。
Enteroendocrine cells (EECs) produce hormones such as glucagon-like peptide 1 and peptide YY that regulate food absorption, insulin secretion, and appetite. Based on the success of glucagon-like peptide 1-based therapies for type 2 diabetes and obesity, EECs are themselves the focus of drug discovery programs to enhance gut hormone secretion. The aim of this study was to identify the transcriptome and peptidome of human EECs and to provide a cross-species comparison between humans and mice. By RNA sequencing of human EECs purified by flow cytometry after cell fixation and staining, we present a first transcriptomic analysis of human EEC populations and demonstrate a strong correlation with murine counterparts. RNA sequencing was deep enough to enable identification of low-abundance transcripts such as G-protein-coupled receptors and ion channels, revealing expression in human EECs of G-protein-coupled receptors previously found to play roles in postprandial nutrient detection. With liquid chromatography-tandem mass spectrometry, we profiled the gradients of peptide hormones along the human and mouse gut, including their sequences and posttranslational modifications. The transcriptomic and peptidomic profiles of human and mouse EECs and cross-species comparison will be valuable tools for drug discovery programs and for understanding human metabolism and the endocrine impacts of bariatric surgery.