ECM stiffness regulates lung fibroblast survival through RasGRF1-dependent signaling

Extracellular matrix stiffness is one of the multiple mechanical signals that alter cellular behavior. During studies exploring the effect of matrix rigidity on lung fibroblast survival, we discovered that enhanced survival on stiff substrates is dependent on elevated Ras activity, owing to the activation of the guanine nucleotide exchange factor, RasGRF1. Mechanistically, we found that the increased Ras activity lead to the activation of both the AKT and ERK pathways. Pharmacological inhibition of AKT or ERK signaling attenuates the elevated survival observed on stiff substrates. AKT signaling regulates the phosphorylation and inactivation of the transcription factor FOXO3a. RNAi experiments demonstrate that FOXO3a activity is critical for the cell death observed on soft substrates. Additionally, downregulation of FOXO3a activity on stiff substrate leads to the degradation of the proapoptotic protein Bim. Depletion of Bim increased the survival of cells on soft substrates. Together, our data show that enhanced matrix stiffness activates a RasGRF1/Ras signaling cascade that regulates the activity of AKT and ERK-dependent FOXO3a and Bim expression to alter cell survival.