TRPV4 modulates substrate stiffness mechanosensing and transcellular pore formation in human Schlemm's canal cells

Pathological changes in the biomechanical environment of Schlemm's canal (SC) inner wall cells, such as substrate stiffening and increased cellular stretch, are associated with ocular hypertension, a key risk factor for the development of glaucoma. Cell membrane stretch can trigger the activation of transient receptor potential vanilloid 4 (TRPV4) mechanosensitive ion channels, allowing calcium influx and initiating downstream signaling. However, the precise role of TRPV4 in SC cell mechanobiology remains unclear. Here, we demonstrate that sustained inhibition of TRPV4 activity modulates substrate stiffness mechanosensing to thereby affect the remodeling of the actin cytoskeleton and extracellular matrix of SC cells. This is accompanied by a reduction in cell stiffness and an increase in transcellular pore forming ability, potentially lowing outflow resistance and risk of ocular hypertension. Conversely, acute activation of TRPV4 channels induces Ca²⁺ influx, increasing transcellular pore formation in SC cells. Notaly, reduced TRPV4 mechanosensing was observed in glaucomatous SC cells, resulting in reduced transcellular pore forming ability. These findings suggest novel potential strategies based on targeting TRPV4 in SC cells for the treatment of ocular hypertension in glaucoma.