Rodriguez-Meira Alba, Buck Gemma, Clark Sally-Ann, Povinelli Benjamin J, Alcolea Veronica, Louka Eleni, McGowan Simon, Hamblin Angela, Sousos Nikolaos, Barkas Nikolaos, Giustacchini Alice, Psaila Bethan, Jacobsen Sten Eirik W, Thongjuea Supat, Mead Adam J
Haematopoietic Stem Cell Biology Laboratory, Medical Research Council Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UK; Medical Research Council Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UK.
Flow Cytometry Facility, Medical Research Council, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UK.
Mol Cell. 2019 Mar 21;73(6):1292-1305.e8. doi: 10.1016/j.molcel.2019.01.009. Epub 2019 Feb 12.
Single-cell RNA sequencing (scRNA-seq) has emerged as a powerful tool for resolving transcriptional heterogeneity. However, its application to studying cancerous tissues is currently hampered by the lack of coverage across key mutation hotspots in the vast majority of cells; this lack of coverage prevents the correlation of genetic and transcriptional readouts from the same single cell. To overcome this, we developed TARGET-seq, a method for the high-sensitivity detection of multiple mutations within single cells from both genomic and coding DNA, in parallel with unbiased whole-transcriptome analysis. Applying TARGET-seq to 4,559 single cells, we demonstrate how this technique uniquely resolves transcriptional and genetic tumor heterogeneity in myeloproliferative neoplasms (MPN) stem and progenitor cells, providing insights into deregulated pathways of mutant and non-mutant cells. TARGET-seq is a powerful tool for resolving the molecular signatures of genetically distinct subclones of cancer cells.
单细胞RNA测序（scRNA-seq）已成为解析转录异质性的强大工具。然而，目前其在癌组织研究中的应用受到绝大多数细胞关键突变热点覆盖不足的阻碍；这种覆盖不足使得无法将同一单细胞的基因和转录读数进行关联。为克服这一问题，我们开发了TARGET-seq，这是一种可从基因组DNA和编码DNA中对单细胞内多个突变进行高灵敏度检测的方法，同时还能进行无偏差的全转录组分析。将TARGET-seq应用于4559个单细胞，我们展示了该技术如何独特地解析骨髓增殖性肿瘤（MPN）干细胞和祖细胞中的转录和基因肿瘤异质性，为突变和非突变细胞的失调途径提供了见解。TARGET-seq是解析癌细胞基因不同亚克隆分子特征的强大工具。
