Targeting miR-184 to Enhance Corneal Epithelial Wound Healing: In Vivo Knockout, Antisense Inhibition, and Exosome-Mediated Delivery.

PURPOSE: MicroRNA-184 (miR-184) is known to negatively regulate corneal epithelial wound healing (CEWH) by modulating target genes. This study aimed to further investigate its role in vivo and explore potential therapeutic strategies targeting miR-184 to promote CEWH.

METHODS

Corneal epithelial-specific Mir184 conditional knockout (cKO) mice were generated using the Cre/loxP system. CEWH was evaluated by fluorescein slit-lamp imaging and histology. Epithelial cell proliferation and cell cycle progression were assessed by in vivo EdU incorporation and flow cytometry, respectively. Planar and vertical cell migration were examined using in vitro scratch assays and an in vivo EdU pulse-chase assay. Expression levels of miR-184 and its targets were measured by RT-qPCR and Western blot analysis. Therapeutic inhibition of miR-184 was achieved through intrastromal locked nucleic acid (LNA)-anti-miR-184 injection, soluble drug-loaded microneedle patches, or exosome-mediated delivery in CEWH mouse models.

RESULTS

Corneal epithelial-specific deletion of miR-184 significantly accelerated CEWH, enhanced epithelial cell proliferation, and promoted cell cycle progression compared with wild-type controls. MiR-184 deletion accelerated corneal epithelial turnover in vivo and enhanced in vitro epithelial cell migration. Loss of miR-184 led to upregulation of target proteins CDC25A, CARM1, and LASP1. Therapeutic inhibition of miR-184 via intrastromal LNA-anti-miR-184, soluble microneedle patches, or exosome-mediated delivery recapitulated the genetic phenotype, promoting CEWH, reducing miR-184 levels, and increasing target protein expression.

CONCLUSIONS

Genetic deletion or therapeutic inhibition of miR-184 promotes CEWH via derepression of target genes. Microneedles and exosome-based delivery represent promising translational approaches to enhance CEWH.