Reprogramming of Pancreatic Exocrine Cells AR42J Into Insulin-producing Cells Using mRNAs for Pdx1, Ngn3, and MafA Transcription Factors

• Published in: Molecular therapy. Nucleic acids 2016, Vol.5 (5), p.e320, Article e320
• Main Authors: Koblas, Tomas, Leontovyc, Ivan, Loukotova, Sarka, Kosinova, Lucie, Saudek, Frantisek
• Format: Article
• Language: English
• Subjects: Cellular biology; diabetes; Insulin; insulin-producing cells; MafA; modified mRNA; Neurogenin 3; Original; Pancreas; Pdx1; reprogramming; Ribonucleic acid; RNA; synthetic mRNA; transdifferentiation; Transplants & implants; β-cells
• ISSN: 2162-2531; 2162-2531
• DOI: 10.1038/mtna.2016.33

Direct reprogramming of pancreatic nonendocrine cells into insulin-producing β-cells represents a promising approach for the treatment of insulin-dependent diabetes. However, its clinical application is limited by the potential for insertional mutagenesis associated with the viral vectors currently used for cell reprogramming. With the aim of developing a nonintegrative reprogramming strategy for derivation of insulin-producing cells, here, we evaluated a new approach utilizing synthetic messenger RNAs encoding reprogramming transcription factors. Administration of synthetic mRNAs encoding three key transcription regulators of β-cell differentiation'Pdx1, Neurogenin3, and MafA'efficiently reprogrammed the pancreatic exocrine cells into insulin-producing cells. In addition to the insulin genes expression, the synthetic mRNAs also induced the expressions of genes important for proper pancreatic β-cell function, including Sur1, Kir6.2, Pcsk1, and Pcsk2. Pretreating cells with the chromatin-modifying agent 5-Aza-2′-deoxycytidine further enhanced reprogramming efficiency, increasing the proportion of insulin-producing cells from 3.5 ± 0.9 to 14.3 ± 1.9% (n = 4). Moreover, 5-Aza-2′-deoxycytidine pretreatment enabled the reprogrammed cells to respond to glucose challenge with increased insulin secretion. In conclusion, our results support that the reprogramming of pancreatic exocrine cells into insulin-producing cells, induced by synthetic mRNAs encoding pancreatic transcription factors, represents a promising approach for cell-based diabetes therapy.