A TRPV4–dependent calcium signaling axis governs lamellipodial actin architecture to promote cell migration

Cell migration is crucial for development and tissue homeostasis, while its dysregulation leads to severe pathologies. Cell migration is driven by the extension of actin–based lamellipodia protrusions, powered by actin polymerization, which is tightly regulated by signaling pathways, including Rho GTPases and Ca ²⁺ signaling. While the importance of Ca ²⁺ signaling in lamellipodia protrusions has been established, the molecular mechanisms linking Ca ²⁺ to lamellipodia assembly are unknown. Here, we identify a novel Ca ²⁺ signaling axis involving the mechano-gated channel TRPV4, which regulates lamellipodia protrusions in various cell types. Using Ca ²⁺ and FRET imaging, we demonstrate that TRPV4-mediated Ca ²⁺ influx upregulates RhoA activity within lamellipodia, which then facilitates formin–mediated actin assembly. Mechanistically, we identify CaMKII and TEM4 as key mediators relaying the TRPV4–mediated Ca ²⁺ signal to RhoA. These data define a molecular pathway by which Ca ²⁺ influx regulates small GTPase activity within a specific cellular domain – lamellipodia – and demonstrate the critical role in organizing the actin machinery and promoting cell migration in diverse biological contexts.