Key Laboratory of Neurological Function and Health & Department of Human Anatomy, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, Guangdong 511436, China.
Department of Pediatrics, University of California-Irvine, Irvine, CA 92697, USA.
Exp Gerontol. 2023 Aug;179:112243. doi: 10.1016/j.exger.2023.112243. Epub 2023 Jun 26.
The differentiation of human induced pluripotent stem cells (hiPSCs) into functional dopaminergic neural precursors is the basis of cell therapy for Parkinson's disease (PD). However, the use of small molecule inhibitors/activators in the differentiation of hiPSCs in vitro leads to cell death and low differentiation efficiency. Moreover, the mechanism of differentiation remains unclear. MiR-210-5p was increased during hiPSCs differentiation. Whether it promotes hiPSCs differentiation and transplantation needs further study. Here, we overexpressed miR-210-5p in hiPSCs to study its roles and mechanisms. We found that miR-210-5p promoted the differentiation of hiPSCs into dopaminergic neural precursors and reduced the expression of SMAD4 and SUFU meanwhile. Luciferase assays showed that miR-210-5p binded to SMAD4 and SUFU, which are key molecules in the key signals (TGF-β and SHH) of hiPSCs differentiation. Furthermore, in the effect evaluation of cell transplantation into parkinsonian rats, the degree of behavioral recovery and the growth of transplanted cells in the group overexpressed miR-210-5p were similar to those in the positive group with all small molecule inhibitors/activators. Therefore, we conclude that miR-210-5p promotes the differentiation of hiPSCs into dopaminergic neural precursors by targeting SMAD4 and SUFU. In the therapeutic evaluation of cell transplantation, miR-210-5p can replace the use of corresponding small molecule inhibitors/activators to reduce cell death. This study provides an experimental basis and a new target for the miRNA-modified differentiation of hiPSCs and cell transplantation in clinical treatment of PD in the future.
人诱导多能干细胞（hiPSCs）向功能性多巴胺能神经前体细胞的分化是帕金森病（PD）细胞治疗的基础。然而，在体外使用小分子抑制剂/激活剂来分化 hiPSCs 会导致细胞死亡和低分化效率。此外，分化的机制尚不清楚。miR-210-5p 在 hiPSCs 分化过程中增加。它是否促进 hiPSCs 分化和移植需要进一步研究。在这里，我们过表达 miR-210-5p 在 hiPSCs 中研究其作用和机制。我们发现 miR-210-5p 促进 hiPSCs 分化为多巴胺能神经前体细胞，同时降低 SMAD4 和 SUFU 的表达。荧光素酶报告基因实验表明，miR-210-5p 与 SMAD4 和 SUFU 结合，SMAD4 和 SUFU 是 hiPSCs 分化的关键信号（TGF-β 和 SHH）中的关键分子。此外，在对帕金森病大鼠进行细胞移植的效果评估中，过表达 miR-210-5p 组的行为恢复程度和移植细胞的生长与所有小分子抑制剂/激活剂的阳性组相似。因此，我们得出结论，miR-210-5p 通过靶向 SMAD4 和 SUFU 促进 hiPSCs 向多巴胺能神经前体细胞分化。在细胞移植的治疗评估中，miR-210-5p 可以替代相应小分子抑制剂/激活剂的使用，以减少细胞死亡。这项研究为未来 miRNA 修饰的 hiPSCs 分化和细胞移植治疗 PD 提供了实验依据和新靶点。
