BioProject

Non-genetic mechanisms have recently emerged as important drivers of therapy failure in cancer, where some cancer cells can enter a reversible drug-tolerant persister state in response to treatment. More...

Non-genetic mechanisms have recently emerged as important drivers of therapy failure in cancer, where some cancer cells can enter a reversible drug-tolerant persister state in response to treatment. While most cancer persisters, like their bacterial counterparts, remain arrested in the presence of drug, a rare subset of cancer persisters can re-enter the cell cycle under constitutive drug treatment. Little is known about the non-genetic mechanisms that enable cancer persisters to simultaneously resist therapy and maintain proliferative capacity in the presence of drug. To address this, we developed Watermelon, a new high-complexity expressed barcode lentiviral library for simultaneous tracing each cell's clonal origin, proliferative state, and transcriptional state, and used it to study this rare, transiently-resistant, proliferative persister population and identify what distinguishes it from non-cycling persisters. Analysis of Watermelon-transduced cancer cell lines demonstrated that cycling and non-cycling persisters arise from different pre-existing cell lineages with distinct transcriptional and metabolic programs. The proliferative capacity of persisters is associated with an upregulation of antioxidant gene programs and a metabolic shift to fatty acid oxidation in specific subpopulations of tumor cells. Overall design: 10X Genomics single cell RNAseq of PC9 cells treated with Osimertinib Lung cancer PC9 cells were treated with osimertinib (300nM) and harvested before treatment, after 3,7, and 14 days of continuous drug treatment. At day 14 prior to sequencing, cells were sorted to three groups based on mCherry expression:cycling (low), moderate cyclers (med) and non-cycling (high) In addition, seven Watermelon lines models from EGFR-driven lung cancer (PC9, HCC827), HER2-driven breast cancer (BT474, EFM192A) and BRAF-driven melanoma (A375, COLO858) and colorectal (HT29) cell lines, treated them with clinically relevant kinase inhibitors for 10 days (300nM osimertinib, 1uM lapatinib or 1uM dabrafenib). At day 10 prior to sequencing, cells were sorted to three groups based om mCherry expression:cycling (low) and non-cycling (high) Less...