Abstract

The cornea is the anterior, transparent tissue of the human eye that serves as its main refractive element. Corneal endothelial cells are arranged as a monolayer on the posterior surface of the cornea and function as a pump to counteract the leakiness of its basement membrane. Maintaining the cornea in a slightly dehydrated state is critical for the maintenance of corneal transparency. Adult human corneal endothelial cells are G1-arrested, even in response to injury, leading to an age-dependent decline in endothelial cell density. Corneal edema and subsequent vision loss ensues when endothelial cell density decreases below a critical threshold. Vision loss secondary to corneal endothelial dysfunction is a common indication for transplantation in developed nations. An impending increase in demand for and a current global shortage of donor corneas will necessitate the development of treatments for vision loss because of endothelial dysfunction that do not rely on donor corneas. Wnt ligands regulate many critical cellular functions, such as proliferation, making them attractive candidates for modulation in corneal endothelial dysfunction. We show that WNT10B causes nuclear transport and binding of RAC1 and β-catenin in human corneal endothelial cells, leading to the activation of Cyclin D1 expression and proliferation. Our findings indicate that WNT10B promotes proliferation in human corneal endothelial cells by simultaneously utilizing both β-catenin-dependent and -independent pathways and suggest that its modulation could be used to treat vision loss secondary to corneal endothelial dysfunction.

Highlights

  • Adult human corneal endothelial cells are G1-arrested, often necessitating transplantation in patients with endothelial dysfunction

  • We provide evidence that WNT10B signals simultaneously through ␤-catenin-dependent and -independent pathways to enhance proliferation in human corneal endothelial cells (CECs), and our results suggest that modulation of WNT10B could be used to treat vision loss secondary to corneal endothelial dysfunction

  • Corneal endothelial cells were transfected at 70% confluence on a 6-well plate with 1.5 ␮M Accell SMARTpool of siRNA targeting p65 (RelA), JUN, Disheveled 2, RAC1, ␤-catenin, or Cyclin D1 (Dharmacon, Pittsburgh, PA) in Accell delivery medium according to the instructions of the manufacturer. 42 h after transfection, the medium was changed to each culture medium and maintained for another 6 h

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Summary

Background

Adult human corneal endothelial cells are G1-arrested, often necessitating transplantation in patients with endothelial dysfunction. Our findings indicate that WNT10B promotes proliferation in human corneal endothelial cells by simultaneously utilizing both ␤-catenin-dependent and -independent pathways and suggest that its modulation could be used to treat vision loss secondary to corneal endothelial dysfunction. We investigated the downstream targets of IL-1␤ capable of driving proliferation in human CECs. Our results show that IL-1␤ activates WNT10B expression and promotes proliferation in human CECs. WNT10B signals through Disheveled and induces independent nuclear transport of RAC1 and ␤-catenin, during which they form a complex to activate expression of Cyclin D1, leading to proliferation of human CECs. We provide evidence that WNT10B signals simultaneously through ␤-catenin-dependent and -independent pathways to enhance proliferation in human CECs, and our results suggest that modulation of WNT10B could be used to treat vision loss secondary to corneal endothelial dysfunction

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