Department of Oncology, University of Cambridge, Hutchison/MRC Research Centre, Hills Road, Cambridge, CB2 0XZ, UK.
Wellcome - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK.
Sci Rep. 2018 Oct 18;8(1):15374. doi: 10.1038/s41598-018-33838-5.
β-cell replacement has been proposed as an effective treatment for some forms of diabetes, and in vitro methods for β-cell generation are being extensively explored. A potential source of β-cells comes from fate conversion of exocrine pancreatic cells into the endocrine lineage, by overexpression of three regulators of pancreatic endocrine formation and β-cell identity, Ngn3, Pdx1 and MafA. Pancreatic ductal organoid cultures have recently been developed that can be expanded indefinitely, while maintaining the potential to differentiate into the endocrine lineage. Here, using mouse pancreatic ductal organoids, we see that co-expression of Ngn3, Pdx1 and MafA are required and sufficient to generate cells that express insulin and resemble β-cells transcriptome-wide. Efficiency of β-like cell generation can be significantly enhanced by preventing phosphorylation of Ngn3 protein and further augmented by conditions promoting differentiation. Taken together, our new findings underline the potential of ductal organoid cultures as a source material for generation of β-like cells and demonstrate that post-translational regulation of reprogramming factors can be exploited to enhance β-cell generation.
β 细胞替代已被提议作为某些类型糖尿病的有效治疗方法，并且正在广泛探索体外生成 β 细胞的方法。β 细胞的一个潜在来源是通过过表达三种调节胰腺内分泌形成和 β 细胞特性的转录因子 Ngn3、Pdx1 和 MafA，将外分泌胰腺细胞命运转化为内分泌谱系。最近已经开发出胰腺导管类器官培养物，可以无限期扩增，同时保持分化为内分泌谱系的潜力。在这里，我们使用小鼠胰腺导管类器官，发现共表达 Ngn3、Pdx1 和 MafA 是生成表达胰岛素且在转录组水平上类似于 β 细胞的细胞所必需且充分的。通过防止 Ngn3 蛋白的磷酸化，可以显著提高 β 样细胞生成的效率，并且通过促进分化的条件进一步增强。总之，我们的新发现强调了导管类器官培养物作为生成 β 样细胞的源材料的潜力，并证明了可以利用重编程因子的翻译后调控来增强 β 细胞的生成。
