Men's Health and Reproductive Health Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Iran Biomed J. 2022 Jul 1;26(4):291-300. doi: 10.52547/ibj.3594.
Recent years have brought notable progress in raising the efficiency of the reprogramming technique so that approaches have evolved from known transgenic factors to only a few miRNAs. Nevertheless, there is a poor understanding of both the key factors and biological networks underlying this reprogramming. The present study aimed to investigate the potential of miR-106b-5p in regulating spermatogonial stem cells (SSCs) to induced pluripotent stem cell (iPSC)-like cells.
We used SSCs because pluripotency is inducible in SSCs under defined culture conditions, and they have a few issues compared to other adult stem cells. As both signaling and post-transcriptional gene controls are critical for pluripotency regulation, we traced the expression of Oct-4, Sox-2, Klf-4, c-Myc, and Nanog (OSKMN). Besides, we considered miR-106b-5p targets using bioinformatic methods.
Our results showed that transfected SSCs with miR-106b-5p increased the expression of the OSKMN factors, which was significantly more than negative control groups. Moreover, using the functional miRNA enrichment analysis, online tools, and databases, we predicted that miR-106b-5p targeted a signaling pathway gene named MAPK1/ERK2, related to regulating stem cell pluripotency.
Together, our data suggest that miR-106b-5p regulates the reprogramming of SSCs into iPSC-like cells. Furthermore, noteworthy progress in the in vitro development of SSCs indicates promise reservoirs and opportunities for future clinical trials.
近年来，提高重编程技术效率的研究取得了显著进展，方法已经从已知的转基因因子演变为仅少数几个 miRNAs。然而，人们对这种重编程的关键因素和生物网络仍知之甚少。本研究旨在探讨 miR-106b-5p 在调节精原干细胞（SSC）向诱导多能干细胞（iPSC）样细胞的潜能。
我们使用 SSC，因为在特定的培养条件下，多潜能性可在 SSC 中诱导，并且与其他成体干细胞相比，它们具有一些问题。由于信号和转录后基因控制对于多潜能性调节至关重要，我们追踪了 Oct-4、Sox-2、Klf-4、c-Myc 和 Nanog（OSKMN）的表达。此外，我们使用生物信息学方法考虑了 miR-106b-5p 的靶标。
我们的结果表明，转染 miR-106b-5p 的 SSC 增加了 OSKMN 因子的表达，明显高于阴性对照组。此外，使用功能 miRNA 富集分析、在线工具和数据库，我们预测 miR-106b-5p 靶向了一个信号通路基因，称为 MAPK1/ERK2，与调节干细胞多潜能性有关。
总之，我们的数据表明，miR-106b-5p 调节 SSC 向 iPSC 样细胞的重编程。此外，SSC 的体外发育方面的显著进展表明，未来的临床试验具有巨大的潜力和机会。
