Deciphering intratumoral heterogeneity using integrated clonal tracking and single-cell transcriptome analyses

Cellular heterogeneity is a major cause of treatment resistance in cancer. Despite recent advances in single-cell genomic and transcriptomic sequencing, it remains difficult to relate measured molecular profiles to the cellular activities underlying cancer. Here, we present an integrated experimental system that connects single cell gene expression to heterogeneous cancer cell growth, metastasis, and treatment response. Our system integrates single cell transcriptome profiling with DNA barcode based clonal tracking in patient-derived xenograft models. We show that leukemia cells exhibiting unique gene expression respond to different chemotherapies in distinct but consistent manners across multiple mice. In addition, we uncover a form of leukemia expansion that is spatially confined to the bone marrow of single anatomical sites and driven by cells with distinct gene expression. Our integrated experimental system can interrogate the molecular and cellular basis of the intratumoral heterogeneity underlying disease progression and treatment resistance. DNA barcoding is a promising technology for the simultaneous analysis of genetic and phenotypic heterogeneity. Here, the authors combine DNA barcoding and single-cell RNA-seq to study heterogeneity, progression and response to therapy in B-cell acute lymphoblastic leukaemia patient-derived xenografts.

Automated summary

Cellular heterogeneity is a major obstacle in cancer treatment, making it difficult to relate molecular profiles to cancer-cell activities. The integrated experimental system presented in this study links single-cell gene expression to cancer cell growth, metastasis, and treatment response, providing insights into intratumoral heterogeneity and disease progression. DNA barcoding combined with single-cell RNA sequencing is a promising approach for analyzing genetic and phenotypic heterogeneity in cancer.