Abstract:
The loss of inner ear hair cells leads to irreversible acoustic injury in mammals, and regeneration of inner ear hair cells to restore hearing loss is challenging. ATOH1 is a key gene in the development and regeneration of hair cells. Small activating RNAs (saRNAs) can target a gene to specifically upregulate its expression. This study aimed to explore whether small activating RNAs could induce the differentiation of human adipose-derived mesenchymal stem cells into hair cell-like cells with a combination of growth factors in vitro and thus provide a new strategy for hair cell regeneration and the treatment of sensorineural hearing loss. Fifteen small activating RNAs targeting the human ATOH1 gene were designed and screened in 293 T and human adipose-derived mesenchymal stem cells, and 3 of these candidates were found to be capable of effectively and stably activating ATOH1 gene expression. The selected small activating RNAs were then transfected into hair cell progenitor cells, and hair cell markers were examined 10 days after transfection. After transfection of the selected small activating RNAs, the expression of the characteristic markers of inner ear hair cells, POU class 4 homeobox 3 (POU4F3) and myosin VIIA (MYO7A), was detected. Human adipose-derived mesenchymal stem cells have the potential to differentiate into human hair cell progenitor cells. In vitro, small activating RNAs were able to induce the differentiation of hair cell progenitor cells into hair cell-like cells. Therefore, RNA activation technology has the potential to provide a new strategy for the regeneration of hair cells.
摘要:
内耳毛细胞的丧失导致哺乳动物不可逆的声损伤,和再生内耳毛细胞恢复听力损失是具有挑战性的。ATOH1是毛细胞发育和再生的症结基因。小激活RNA(saRNA)可以靶向基因以特异性上调其表达。本研究旨在探讨小活化RNA是否能在体外联合生长因子诱导人脂肪间充质干细胞分化为毛细胞样细胞,从而为毛细胞再生和感音神经性耳聋的治疗提供新的策略。在293T和人脂肪间充质干细胞中设计并筛选了15种靶向人ATOH1基因的小活化RNA,发现这些候选物中的3个能够有效且稳定地激活ATOH1基因表达。然后将选择的小活化RNA转染到毛细胞祖细胞中,转染后10天检查毛细胞标记物。转染选定的小活化RNA后,内耳毛细胞特征性标志物的表达,POU类4homeobox3(POU4F3)和肌球蛋白VIIA(MYO7A),被检测到。人脂肪间充质干细胞具有向人毛细胞祖细胞分化的潜能。体外,小活化RNA能够诱导毛细胞祖细胞分化为毛细胞样细胞.因此,RNA激活技术有潜力为毛细胞的再生提供新的策略。