Human corneal endothelial cells (hCECs) are restricted in proliferative capacity in vivo. Reduction in the number of
hCEC leads to persistent
corneal edema requiring
corneal transplantation. This study demonstrates the functions of
SIRT1 in hCECs and its potential for corneal endothelial regeneration. Cell morphology, cell growth rates and proliferation-associated
proteins were compared in normal and senescent hCECs.
SIRT1 was activated using the CRISPR/dCas9 activation system (SIRT1a). The plasmids were transfected into CECs of six-week-old Sprague-Dawley rats using electroporation and cryoinjury was performed. Senescent cells were larger, elongated and showed lower proliferation rates and lower
SIRT1 levels.
SIRT1 activation promoted the wound healing of CECs. In vivo transfection of SIRT1a promoted the regeneration of CECs. The proportion of the S-phase cells was lower in senescent cells and elevated upon SIRT1a activation.
SIRT1 regulated cell proliferation, proliferation-associated
proteins, mitochondrial membrane potential, and oxidative stress levels. In conclusion, corneal endothelial senescence is related with a decreased
SIRT1 level. SIRT1a promotes the regeneration of CECs by inhibiting
cytokine-induced cell death and senescence. Gene function activation
therapy using SIRT1a may serve as a novel treatment strategy for
hCEC diseases.