MiR-381-3p Cooperated With Hes1 to Regulate the Proliferation and Differentiation of Retinal Progenitor Cells.

Xiaochan Dai,Jiajing Wang,Yiqi Wang,Moxin Chen,Jing Ji,Dandan Zhang,Yahan Ju,Ni Ni,Na Sun,Zhimin Tang,Ping Gu

doi:10.3389/fcell.2022.853215

Abstract

Retinal progenitor cells (RPCs) transplantation has become a promising therapy for retinal degeneration, which is a major kind of ocular diseases causing blindness. Since RPCs have limited proliferation and differentiation abilities toward retinal neurons, it is urgent to resolve these problems. MicroRNAs have been reported to have vital effects on stem cell fate. In our study, the data showed that overexpression of miR-381-3p repressed Hes1 expression, which promoted RPCs differentiation, especially toward neuronal cells, and inhibited RPCs proliferation. Knockdown of endogenous miR-381-3p increased Hes1 expression to inhibit RPCs differentiation and promote proliferation. In addition, a luciferase assay demonstrated that miR-381-3p directly targeted the Hes1 3’ untranslated region (UTR). Taken together, our study demonstrated that miR-381-3p regulated RPCs proliferation and differentiation by targeting Hes1, which provides an experimental basis of RPCs transplantation therapy for retinal degeneration.

Highlights

Retinal degeneration is a primary kind of blindness-causing ocular disease worldwide, including agerelated macular degeneration and retinitis pigmentosa, which can cause irreversible blindness in patients (Bourne et al, 2013)
To determine the role that miR-381 plays in Retinal progenitor cells (RPCs), we first evaluated the endogenous expression levels of miR-381-3p during RPCs differentiation. qPCR analysis showed that miR-381-3p expression increased dramatically with the process of RPCs differentiation, which reached approximately 200-fold at the seventh day compared to the expression at the beginning (Figure 1A)
To investigate whether miR-381-3p had an effect on RPCs proliferation and differentiation, RPCs were transfected with miR381-3p mimics or inhibitors and cultured in proliferation or differentiation medium. miR-381-3p expression in the pre-miR-3813p group was increased nearly 150-fold, while that in the anti-miR381-3p group was decreased nearly 0.002-fold compared to that in the controls, showing great transfection efficiency (Figure 2A)

Summary

Introduction

Retinal degeneration is a primary kind of blindness-causing ocular disease worldwide, including agerelated macular degeneration and retinitis pigmentosa, which can cause irreversible blindness in patients (Bourne et al, 2013). Retinal progenitor cells (RPCs), which have been discovered in adult mammalian eyes and even successfully extracted from the human retina, have the potential for self-renewal and differentiation toward retinal neuronal and glial lineages (Tropepe et al, 2000; Coles et al, 2004; Xia et al, 2012). The limited ability of RPCs to proliferate and differentiate toward retinal neurons hinders their future clinical application (Gu et al, 2007). MicroRNAs play essential roles in many biological processes, such as metabolism, development, proliferation, apoptosis, and stem cell differentiation (Krol et al, 2010). A few microRNAs have been identified that can regulate RPCs proliferation and differentiation. MiR-17 and miR-29a can reduce RPCs proliferation and promote differentiation (Zhang et al, 2017; Sun et al, 2020). MiR-762 can promote RPCs proliferation and inhibit miR-381-3p Regulate RPCs Fate differentiation (Gao et al, 2020). The role microRNAs play in RPCs fate is not thoroughly understood

Methods

Results

Conclusion