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.
Journal article
Mol Cell
21/03/2019
73
1292 - 1305.e8
Heterogeneity, TARGET-seq, cancer, hematopoiesis, leukemia, mutations, myeloproliferative neoplasm, sequencing, single-cell, transcriptomics, Biomarkers, Tumor, DNA Mutational Analysis, Genetic Heterogeneity, High-Throughput Nucleotide Sequencing, Humans, Jurkat Cells, K562 Cells, Leukemia, Mutation, Reproducibility of Results, Schizosaccharomyces, Sequence Analysis, RNA, Single-Cell Analysis