Single-cell morphodynamical trajectories enable prediction of gene expression accompanying cell state change

Extracellular signals induce changes to molecular programs that modulate multiple cellular phenotypes, including proliferation, motility, and differentiation status. The connection between dynamically adapting phenotypic states and the molecular programs that define them is not well understood. Here we develop data-driven models of single-cell phenotypic responses to extracellular stimuli by linking gene transcription levels to "morphodynamics" - changes in cell morphology and motility observable in time-lapse image data. We adopt a dynamics-first view of cell state by grouping single-cell trajectories into states with shared morphodynamic responses. The single-cell trajectories enable development of a first-of-its-kind computational approach to map live-cell dynamics to snapshot gene transcript levels, which we term MMIST, Molecular and Morphodynamics-Integrated Single-cell Trajectories. The key conceptual advance of MMIST is that cell behavior can be quantified based on dynamically defined states and that extracellular signals change the overall distribution of cell states by altering rates of switching between states. We find a cell state landscape that is bound by epithelial and mesenchymal endpoints, with distinct sequences of epithelial to mesenchymal transition (EMT) and mesenchymal to epithelial transition (MET) intermediates. The analysis yields predictions for gene expression changes consistent with curated EMT gene sets and predicts expression of thousands of RNA transcripts through extracellular signal-induced EMT and MET with near-continuous time resolution. The MMIST framework leverages true single-cell dynamical behavior to generate molecular-level omics inferences and is broadly applicable to other biological domains, time-lapse imaging approaches and molecular snapshot data. SummaryIn normal homeostatic tissues, extracellular signals induce changes in the behavior and state of epithelial cells, and aberrant responses to such signals are associated with diseases. To decode and potentially steer these responses, it is essential to link live-cell behavior to molecular programs; however, high-throughput molecular techniques are destructive or require fixation. Here we present a novel computational approach to connect single-cell measures of cell phenotype and behavior to bulk molecular readouts, enabling prediction of dynamic changes in gene expression programs. This reveals molecular programs associated with distinct cell states and identifies drivers that may be manipulated to control cell state change.