Cargando…
Reprogramming of Pancreatic Exocrine Cells AR42J Into Insulin-producing Cells Using mRNAs for Pdx1, Ngn3, and MafA Transcription Factors
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...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5014516/ https://www.ncbi.nlm.nih.gov/pubmed/27187823 http://dx.doi.org/10.1038/mtna.2016.33 |
_version_ | 1782452302592868352 |
---|---|
author | Koblas, Tomas Leontovyc, Ivan Loukotova, Sarka Kosinova, Lucie Saudek, Frantisek |
author_facet | Koblas, Tomas Leontovyc, Ivan Loukotova, Sarka Kosinova, Lucie Saudek, Frantisek |
author_sort | Koblas, Tomas |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5014516 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-50145162016-09-19 Reprogramming of Pancreatic Exocrine Cells AR42J Into Insulin-producing Cells Using mRNAs for Pdx1, Ngn3, and MafA Transcription Factors Koblas, Tomas Leontovyc, Ivan Loukotova, Sarka Kosinova, Lucie Saudek, Frantisek Mol Ther Nucleic Acids Original Article 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. Nature Publishing Group 2016-05 2016-05-17 /pmc/articles/PMC5014516/ /pubmed/27187823 http://dx.doi.org/10.1038/mtna.2016.33 Text en Copyright © 2016 Official journal of the American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by-nc-sa/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/ |
spellingShingle | Original Article Koblas, Tomas Leontovyc, Ivan Loukotova, Sarka Kosinova, Lucie Saudek, Frantisek Reprogramming of Pancreatic Exocrine Cells AR42J Into Insulin-producing Cells Using mRNAs for Pdx1, Ngn3, and MafA Transcription Factors |
title | Reprogramming of Pancreatic Exocrine Cells AR42J Into Insulin-producing Cells Using mRNAs for Pdx1, Ngn3, and MafA Transcription Factors |
title_full | Reprogramming of Pancreatic Exocrine Cells AR42J Into Insulin-producing Cells Using mRNAs for Pdx1, Ngn3, and MafA Transcription Factors |
title_fullStr | Reprogramming of Pancreatic Exocrine Cells AR42J Into Insulin-producing Cells Using mRNAs for Pdx1, Ngn3, and MafA Transcription Factors |
title_full_unstemmed | Reprogramming of Pancreatic Exocrine Cells AR42J Into Insulin-producing Cells Using mRNAs for Pdx1, Ngn3, and MafA Transcription Factors |
title_short | Reprogramming of Pancreatic Exocrine Cells AR42J Into Insulin-producing Cells Using mRNAs for Pdx1, Ngn3, and MafA Transcription Factors |
title_sort | reprogramming of pancreatic exocrine cells ar42j into insulin-producing cells using mrnas for pdx1, ngn3, and mafa transcription factors |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5014516/ https://www.ncbi.nlm.nih.gov/pubmed/27187823 http://dx.doi.org/10.1038/mtna.2016.33 |
work_keys_str_mv | AT koblastomas reprogrammingofpancreaticexocrinecellsar42jintoinsulinproducingcellsusingmrnasforpdx1ngn3andmafatranscriptionfactors AT leontovycivan reprogrammingofpancreaticexocrinecellsar42jintoinsulinproducingcellsusingmrnasforpdx1ngn3andmafatranscriptionfactors AT loukotovasarka reprogrammingofpancreaticexocrinecellsar42jintoinsulinproducingcellsusingmrnasforpdx1ngn3andmafatranscriptionfactors AT kosinovalucie reprogrammingofpancreaticexocrinecellsar42jintoinsulinproducingcellsusingmrnasforpdx1ngn3andmafatranscriptionfactors AT saudekfrantisek reprogrammingofpancreaticexocrinecellsar42jintoinsulinproducingcellsusingmrnasforpdx1ngn3andmafatranscriptionfactors |