英国伦敦大学癌症研究所Christopher J. Tape研究团队开发了类器官内细胞特异性信号网络分析的新方法。相关论文在线发表在2020年2月17日出版的《自然-方法学》杂志上。

研究人员对类器官和类器官共培养中转录后修饰（PTM）网络进行了多变量单细胞分析。通过对来自小肠类器官> 1百万个单细胞中28个PTM进行的质谱分析，研究人员揭示了干细胞、Paneth细胞、肠内分泌细胞、簇和杯状细胞以及肠上皮细胞中特定细胞类型和细胞状态的信号网络。

将单细胞PTM分析与巯基反应性类器官原位条形码（TOBis）集成在一起，可实现培养类器官之间信号网络的高通量比较。大肠癌类器官共培养的细胞类型特异性PTM分析显示，shApc、Kras^G12D和Trp53^R172H细胞自模拟信号传导状态通常是由基质成纤维细胞和巨噬细胞所诱导的。

这些结果揭示了如何修改标准的大规模流式细胞工作流程，以实现健康和癌症类器官的高通量多元细胞类型特异性信号分析。

据介绍，尽管类器官被广泛用作仿生组织模型，但仍缺乏用于全面分析类器官中特定细胞类型翻译后修饰（PTM）信号网络的方法。

附：英文原文

Title: Cell-type-specific signaling networks in heterocellular organoids

Author: Xiao Qin, Jahangir Sufi, Petra Vlckova, Pelagia Kyriakidou, Sophie E. Acton, Vivian S. W. Li, Mark Nitz, Christopher J. Tape

Issue&Volume: 2020-02-17

Abstract: 

Despite the widespread adoption of organoids as biomimetic tissue models, methods to comprehensively analyze cell-type-specific post-translational modification (PTM) signaling networks in organoids are absent. Here, we report multivariate single-cell analysis of such networks in organoids and organoid cocultures. Simultaneous analysis by mass cytometry of 28 PTMs in >1 million single cells derived from small intestinal organoids reveals cell-type- and cell-state-specific signaling networks in stem, Paneth, enteroendocrine, tuft and goblet cells, as well as enterocytes. Integrating single-cell PTM analysis with thiol-reactive organoid barcoding in situ (TOBis) enables high-throughput comparison of signaling networks between organoid cultures. Cell-type-specific PTM analysis of colorectal cancer organoid cocultures reveals that shApc, KrasG12D and Trp53R172H cell-autonomously mimic signaling states normally induced by stromal fibroblasts and macrophages. These results demonstrate how standard mass cytometry workflows can be modified to perform high-throughput multivariate cell-type-specific signaling analysis of healthy and cancerous organoids.

DOI: 10.1038/s41592-020-0737-8

Source: https://www.nature.com/articles/s41592-020-0737-8