Samaneh Arab, Mohammad-Reza Mahmoudian-Sani , Najmeh Fattahi , Zakiye Ekhlasi, Samira Asgharzade,
Volume 83, Issue 5 (August 2025)
Abstract
Background: Retinal photoreceptor degeneration is a major cause of blindness. Stem cell therapies offer promise, and the miR-183/96/182 cluster, particularly miR-182 and miR-183, plays a crucial role in photoreceptor development and survival. Targeting these miRNAs may enhance human bone marrow–derived mesenchymal stem cells) hBMSCs (differentiation into photoreceptor-like cells, improving their therapeutic potential.
Methods: This in vitro study was conducted from April 2019 to March 2021 at the Clinical Biochemistry Research Center, Shahrekord University of Medical Sciences. hBMSCs were cultured in DMEM with fetal bovine serum and transfected with miR-182 and miR-183 mimics using Lipofectamine, with a scramble miRNA control. Transfection efficiency and miRNA overexpression were evaluated at 24 and 48 hours using real-time PCR. miRNA expression was normalised to Snord, while mRNA levels were normalised to GAPDH using the 2−ΔΔCt method. Photoreceptor-like differentiation was assessed by measuring the expression of retina-specific transcription factors and markers (OTX2, CRX, NRL, SLC1A1, PKCα, Recoverin, and RHO). Statistical analyses included the Shapiro–Wilk test for normality and the Mann-Whitney U test for group comparisons. Data were reported as Mean ± SEM, with 95% confidence intervals, and significance set at α = 0.05.
Results: Transfection of miR-182 and miR-183 significantly increased miRNA levels at 24–48 hours (P < 0.001) compared to the scramble control. This led to a marked upregulation of retinal-related genes, including CRX, OTX2, PKCα, Recoverin, NRL, and RHO, indicating activation of the photoreceptor gene network. Time-resolved analysis revealed stronger effects at 24–48 hours, supporting a transient window for pro-differentiation. RHO and CRX exhibited the most significant increases, while OTX2 and PKCα showed parallel rises, suggesting coordinated activation of early and intermediate photoreceptor programs. Scramble controls did not show comparable changes.
Conclusion: Transient overexpression of miR-182 and miR-183 in hBMSCs activates a photoreceptor-like gene expression program, promoting differentiation toward photoreceptor-like cells. This finding supports the potential use of miR-182/183 in stem cell-based therapies for retinal degeneration. Further studies should confirm protein expression, functional outcomes, and in vivo efficacy.
