Matrix Stiffness Induces Endothelial Network Senescence

ABSTRACT Identifying the drivers of cellular senescence that contribute to the decline in vascular function with age and disease is critical for developing restorative interventions. Here, we investigated how increased mechanical stress from extracellular matrix (ECM) stiffening shapes endothelial cell (EC) senescence. We developed a 3D human in vitro model that decouples mechanical stress from inflammatory or biochemical signals, enabling the study of senescence responses to tissue stiffening alone. We found that matrix stiffening induces an EC senescence phenotype with elevated p16/p21 and an immunomodulatory senescence‐associated secretory phenotype (SASP), in the absence of inflammatory signals. This mechano‐induced senescence activates Notch signaling, and treatment with an FDA‐approved γ‐secretase inhibitor attenuates stiffness‐induced senescence. Analysis of fibrotic capsule tissue from patients with synthetic breast implants, a model of localized, mechanically driven fibrosis, validated an increase in p16 ⁺ Notch1 ⁺ endothelial populations. Complementary single‐cell RNA sequencing data further confirmed enrichment of Notch‐ and SASP‐related gene programs. Our work provides a human‐relevant platform for studying targetable stages of endothelial mechanoaging and identifies potential therapeutic targets associated with stiffness‐induced endothelial senescence for mechanically remodeled tissues.