Abstract
Purpose :
The secretome of corneal stromal cells (keratocytes, KCs) has been demonstrated to enhance proliferation of corneal cells and promote corneal healing. The components of secretome include exosomes (which contain miRNAs). We propose that injection of exosomes or transfection of miRNA can be utilized as a novel therapeutic approach for corneal injuries.
Methods :
The proliferative (growth-promoting) effects of cell secretions, exosomes and miRNA were verified by the culture of human corneal endothelial cells (B4G12 cells) and primary human corneal endothelial cells (HCECs). Exosomes were identified and quantified by a nanoparticle tracking analyzer (NTA) and transmission electron microscopy (TEM). The identification and detection of miRNAs were performed by RNA-seq and reverse transcriptase PCR (RT-PCR). The effects of exosomes and miRNA on the cell cycle were verified by flow cytometry and immunoblotting. The ability of exosomes and miRNA to help damaged corneas heal was tested in a liquid nitrogen corneal damage rabbit model.
Results :
We found that KC-derived exosomes enhance the proliferation of corneal cells. Furthermore, lysophosphatidic acid (LPA) treatment significantly increased KC exosome-derived miR-221-3p, which enhanced the proliferative effect of corneal cells. It was also confirmed that intrastromal injection of KC exosomes and miR-221-3p promotes corneal healing.
Conclusions :
We believe that stromal injection of exosomes or the transformational expression of miR-221-3p have potential applications in the treatment of corneal injury.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.