美国华盛顿大学Anoop P. Patel和美国弗雷德·哈钦森癌症研究中心Steven Henikoff研究团队合作取得最新进展。他们建立染色质修饰在分化和肿瘤进展中的单细胞CUT＆Tag分析技术。该研究于2021年4月12日发表于国际一流学术期刊《自然-生物技术》。

他们将CUT＆Tag方法调整为可扩展的基于纳孔和基于液滴的单细胞平台，以描绘复杂组织中的单细胞（scCUT＆Tag）以及人类胚胎干细胞分化过程中的染色质景观。他们专注于分析在单个细胞中以组蛋白H3 Lys27三甲基化（H3K27me3）为标记的多梳基团（PcG）沉默区域，作为染色质可及性的正交方法来鉴定细胞状态。

他们显示H3K27me3的scCUT＆Tag分析可区分人血中的细胞类型，并允许从异质组织生成特定细胞类型的PcG景观。此外，他们使用scCUT＆Tag对治疗前后脑肿瘤患者的H3K27me3进行了分析，鉴定了肿瘤微环境中的细胞类型以及主要样品和治疗后PcG活性的异质性。

据了解，定量建立单细胞中基因表达和染色质可及性的方法已经建立，但是具有特定组蛋白修饰的染色质区域的单细胞分析在技术上具有挑战性。

附：英文原文

Title: Single-cell CUT&Tag analysis of chromatin modifications in differentiation and tumor progression

Author: Steven J. Wu, Scott N. Furlan, Anca B. Mihalas, Hatice S. Kaya-Okur, Abdullah H. Feroze, Samuel N. Emerson, Ye Zheng, Kalee Carson, Patrick J. Cimino, C. Dirk Keene, Jay F. Sarthy, Raphael Gottardo, Kami Ahmad, Steven Henikoff, Anoop P. Patel

Issue&Volume: 2021-04-12

Abstract: Methods for quantifying gene expression1 and chromatin accessibility2 in single cells are well established, but single-cell analysis of chromatin regions with specific histone modifications has been technically challenging. In this study, we adapted the CUT&Tag method3 to scalable nanowell and droplet-based single-cell platforms to profile chromatin landscapes in single cells (scCUT&Tag) from complex tissues and during the differentiation of human embryonic stem cells. We focused on profiling polycomb group (PcG) silenced regions marked by histone H3 Lys27 trimethylation (H3K27me3) in single cells as an orthogonal approach to chromatin accessibility for identifying cell states. We show that scCUT&Tag profiling of H3K27me3 distinguishes cell types in human blood and allows the generation of cell-type-specific PcG landscapes from heterogeneous tissues. Furthermore, we used scCUT&Tag to profile H3K27me3 in a patient with a brain tumor before and after treatment, identifying cell types in the tumor microenvironment and heterogeneity in PcG activity in the primary sample and after treatment.

DOI: 10.1038/s41587-021-00865-z

Source: https://www.nature.com/articles/s41587-021-00865-z