Abstract
A cell's ability to sense and respond to the mechanical properties of the extracellular matrix (ECM) is essential for maintaining tissue homeostasis, and its disruption contributes to diseases such as fibrosis, cardiovascular disorders, and cancer. Effective mechanical coupling between the plasma membrane, the underlying filamentous actin (F-actin) cytoskeleton, and integrin-based adhesion complexes (IACs) is required to link ECM mechanics to cell morphology, yet the underlying mechanisms remain incompletely understood. Here, we combine computational modeling and high-resolution imaging to show that integrin-ECM bonds determine F-actin cytoskeleton organization. On soft substrates, short-lived IACs bonds allow rapid actin retrograde flow and dense branching, restricting protrusion and limiting cell spreading. In contrast, stiff substrates or Mn²⁺-mediated integrin activation stabilize adhesions, promote filament alignment, and drive membrane protrusion for cell spreading. These cytoskeletal transitions arise from feedback between adhesion strength and the spatial positioning of the F-actin barbed ends relative to the leading-edge membrane. This positioning determines whether filaments polymerize into linear bundles or branch into dendritic networks, each generating distinct protrusive forces that regulate cell spreading. Collectively, our findings establish integrin-ECM bond stability as a key regulator of F-actin cytoskeleton organization and cell morphology.