Arterial stiffening, a major cardiovascular risk factor, is driven by aberrant mechanotransduction in vascular smooth muscle cells (VSMCs), yet the critical mechanoreceptors and underlying mechanisms remain elusive. Here, we identified Ephrin receptor A2 (EphA2) as a significantly upregulated mechanosensitive receptor in stiffened arteries from a 5/6 nephrectomy mouse model. Genetic deletion of Epha2 in VSMCs markedly attenuated arterial stiffening. Utilizing polyacrylamide gels of varying stiffness and in situ stiffening bioclick hydrogels, we demonstrated that matrix stiffening directly induces EphA2 phase separation, forming a biomolecular condensate that serves as a signaling hub to recruit and activate ERK1/2. This leads to phosphorylation of the transcription factor CREB and subsequent upregulation of the pro-remodeling nuclear receptor NR4A3. To translate this discovery, we designed a retro-reversed peptide targeting the intrinsically disordered regions (IDRs) of EphA2, which effectively disrupted phase separation and mitigated VSMCs dysfunction in vitro. Crucially, in vivo delivery of this peptide via VAPG-modified nanoparticles significantly alleviated arterial calcification and stiffening in mice. Our study establishes EphA2 phase separation as a pivotal mechanism in vascular mechanotransduction and unveils a novel EphA2-ERK1/2-NR4A3 signaling axis, thereby presenting a promising therapeutic strategy for combating arterial stiffening by targeting pathological biomolecular condensates.
Keywords: Arterial stiffening; EphA2; Matrix stiffness; Phase separation; Vascular smooth muscle.
© 2026 The Authors.