Zhao Feng, Liu Xiaoyu, Wang Zhe, Lang Hongxin, Zhang Tao, Wang Rui, Lin Xuewen, He Dan, Shi Ping, Pang Xining
Department of Stem Cells and Regenerative Medicine, Shenyang Key Laboratory for Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology, Ministry of Public Health, and Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical University, 77 Puhe Street, Shenbei New District, Shenyang City 110122, Liaoning Province, China.
Department of Obstetrics and Gynecology, Center for Assisted Reproduction, Shengjing Hospital of China Medical University, 39 Huaxiang Street, Tiexi District, Shenyang City 110022, Liaoning Province, China.
Mol Ther Nucleic Acids. 2020 Mar 6;19:1110-1122. doi: 10.1016/j.omtn.2020.01.001. Epub 2020 Jan 16.
MicroRNAs (miRNAs) that play key roles in the generation of insulin-producing cells from stem cells provide a cell-based approach for insulin replacement therapy. In this study, we used next-generation sequencing to detect the miRNA expression profile of normal mouse pancreatic β cells, non-β cells, bone marrow mesenchymal stem cells (BM-MSCs), and adipose-derived stem cells (ADSCs) and determined relative miRNA expression levels in mouse pancreatic β cells. After the novel mouse miRNA candidates were identified using miRDeep 2.0, we found that Chr13_novelMiR7354-5p, a novel miRNA candidate, significantly promoted the differentiation of BM-MSCs into insulin-producing cells in vitro. Furthermore, Chr13_novelMiR7354-5p-transfected BM-MSCs reversed hyperglycemia in streptozotocin (STZ)-treated diabetic mice. In addition, bioinformatics analyses, a luciferase reporter assay, and western blotting demonstrated that Chr13_novelMiR7354-5p targeted Notch1 and Rbpj. Our results provide compelling evidence of the existence of 65 novel mouse miRNA candidates and present a new treatment strategy to generate insulin-producing cells from stem cells.
在干细胞生成胰岛素分泌细胞过程中发挥关键作用的微小RNA（miRNA）为胰岛素替代疗法提供了一种基于细胞的方法。在本研究中，我们使用新一代测序技术检测正常小鼠胰腺β细胞、非β细胞、骨髓间充质干细胞（BM-MSC）和脂肪来源干细胞（ADSC）的miRNA表达谱，并确定小鼠胰腺β细胞中的相对miRNA表达水平。使用miRDeep 2.0鉴定出新的小鼠miRNA候选物后，我们发现一种新的miRNA候选物Chr13_novelMiR7354-5p在体外显著促进BM-MSC向胰岛素分泌细胞的分化。此外，用Chr13_novelMiR7354-5p转染的BM-MSC可逆转链脲佐菌素（STZ）处理的糖尿病小鼠的高血糖症。另外，生物信息学分析、荧光素酶报告基因检测和蛋白质印迹表明Chr13_novelMiR7354-5p靶向Notch1和Rbpj。我们的结果为65种新的小鼠miRNA候选物的存在提供了有力证据，并提出了一种从干细胞生成胰岛素分泌细胞的新治疗策略。
