Cell cycling throttles epithelial--mesenchymal transition

Abstract

Cell cycle phase is an understudied source of variation in gene expression. EMT is a case in point: this cell state transition that drives metastasis has been studied widely across tissue, stimulus, and microenvironmental conditions, yet cell cycle effects are routinely regressed out of single-cell EMT analyses. Here, we compare EMT between G1 and cycling (S/G2/M) cell states in two independent scRNA-seq datasets of TGF-{beta}-stimulated MCF10A mammary epithelial cells (varying dose and timepoint). G1 cells consistently show higher expression of EMT genes and upregulated RNA velocities than cycling cells, with the strongest upregulation in G1 EMT intermediate states. Consistent with the go-or-grow hypothesis, EMT and cell cycle programs inversely correlate; moreover, this anticorrelation extends to RNA velocity and strengthens with TGF-{beta} dose. We identify a 77-gene G1-specific EMT signature enriched for actin cytoskeletal and extracellular matrix genes that would otherwise be lost by regressing out cell cycle effects. This gene set prompts the hypothesis that, unconstrained by the cytoskeletal demands of mitosis, G1 cells have greater capacity to execute EMT. Further, we developed an EMT gene regulatory model coupled with a cell cycle module, and found through simulation that the model recapitulated this inverse EMT--cell cycle relationship. Using this model, we predict that cycle-selective chemotherapy enriches for the G1 intermediate state, which has high metastatic potential. Cell cycle phase thus ought not be regressed out of EMT states, given its influence on EMT.