Negative pressure mechanical signal increases the phosphorylation of eNOS Ser1177 by upregulating HSP90 expression to promote wound angiogenesis.

Background: Chronic wound pathogenesis involves impaired angiogenesis. While negative pressure wound therapy (NPWT) clinically promotes angiogenesis, its biomechanical mechanisms remain unclear.

Methods: A mechanical stretching model simulating NPWT was established in vitro. Multiomics approaches (single-cell sequencing, Ch-IP, Co-IP, and molecular docking) were employed to dissect HSP90-related regulatory networks. Typical molecular biological techniques are used to detect the expression of relevant proteins. Moreover, a rat dorsal wound model was used for the animal experiments.

Results: NPWT-induced mechanical stimulation activates the GNAS/CREB1/HSP90 axis, increasing HSP90 transcription via CREB1 nuclear translocation. Elevated HSP90 displaces Cav-1 to augment eNOS Ser1177 phosphorylation, driving angiogenesis to promote wound healing. Pharmacological or genetic disruption of GNAS/CREB1 suppresses HSP90 expression and angiogenic capacity.

Conclusion: This study reveals a GNAS-mediated mechanotransduction pathway that activates HSP90-dependent eNOS signaling to accelerate wound angiogenesis, suggesting novel targets for therapeutic intervention.