A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in CD34+ Cells
Upon functional loss of insulin producing islet β-cells, some patients with diabetes become dependent on life-long insulin supplementation therapy. Bioengineering surrogate insulin producing cells is an alternative replacement strategy. We have developed a novel approach using short-activating RNA o...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2013-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253116301561 |
| _version_ | 1818137296213377024 |
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| author | Vikash Reebye Pål Sætrom Paul J Mintz John J Rossi Noriyuki Kasahara Georgios Nteliopoulos Joanna Nicholls Abdelali Haoudi Myrtle Gordon Nagy A Habib |
| author_facet | Vikash Reebye Pål Sætrom Paul J Mintz John J Rossi Noriyuki Kasahara Georgios Nteliopoulos Joanna Nicholls Abdelali Haoudi Myrtle Gordon Nagy A Habib |
| author_sort | Vikash Reebye |
| collection | DOAJ |
| description | Upon functional loss of insulin producing islet β-cells, some patients with diabetes become dependent on life-long insulin supplementation therapy. Bioengineering surrogate insulin producing cells is an alternative replacement strategy. We have developed a novel approach using short-activating RNA oligonucleotides to differentiate adult human CD34+ cells into insulin-secreting cells. By transfecting RNA to increase transcript levels of the master regulator of insulin biosynthesis, v-maf musculoaponeurotic fibrosarcoma oncogene homolog A (MafA), several pancreatic endodermal genes were upregulated during the differentiation procedure. These included Pancreatic and duodenal homeobox gene-1 (PDX1), Neurogenin 3, NeuroD, and NK6 homeobox 1 (NKx6-1). Differentiated CD34+ cells also expressed glucokinase, glucagon-like peptide 1 receptor (GLP1R), sulfonylurea receptor-1 (SUR1) and phogrin'all essential for glucose sensitivity and insulin secretion. The differentiated cells appropriately processed C-peptide and insulin in response to increasing glucose stimulation as shown by enzyme-linked immunosorbent assay (ELISA), fluorescence-activated cell sorting analysis, western blotting, and immunofluorescence staining. We provide a new approach using short-activating RNA in developing insulin producing surrogate cells for treating diabetes. |
| first_indexed | 2024-12-11T09:54:03Z |
| format | Article |
| id | doaj.art-d2178392d1054a4c997cfda696da30bf |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-11T09:54:03Z |
| publishDate | 2013-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-d2178392d1054a4c997cfda696da30bf2022-12-22T01:12:19ZengElsevierMolecular Therapy: Nucleic Acids2162-25312013-01-012C10.1038/mtna.2013.23A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in CD34+ CellsVikash Reebye0Pål Sætrom1Paul J Mintz2John J Rossi3Noriyuki Kasahara4Georgios Nteliopoulos5Joanna Nicholls6Abdelali Haoudi7Myrtle Gordon8Nagy A Habib9Department of Surgery and Cancer, Imperial College London, London, UKDepartment of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, NorwayDepartment of Surgery and Cancer, Imperial College London, London, UKDivision of Molecular Biology, Beckman Research Institute of City of Hope, Duarte, California, USADepartment of Medicine, UCLA School of Medicine, Los Angeles, California, USADepartment of Hematology, Faculty of Medicine, Imperial College London, London, UKDepartment of Surgery and Cancer, Imperial College London, London, UKQatar Biomedical Research Institute, Education City, Doha, QatarDepartment of Hematology, Faculty of Medicine, Imperial College London, London, UKDepartment of Surgery and Cancer, Imperial College London, London, UKUpon functional loss of insulin producing islet β-cells, some patients with diabetes become dependent on life-long insulin supplementation therapy. Bioengineering surrogate insulin producing cells is an alternative replacement strategy. We have developed a novel approach using short-activating RNA oligonucleotides to differentiate adult human CD34+ cells into insulin-secreting cells. By transfecting RNA to increase transcript levels of the master regulator of insulin biosynthesis, v-maf musculoaponeurotic fibrosarcoma oncogene homolog A (MafA), several pancreatic endodermal genes were upregulated during the differentiation procedure. These included Pancreatic and duodenal homeobox gene-1 (PDX1), Neurogenin 3, NeuroD, and NK6 homeobox 1 (NKx6-1). Differentiated CD34+ cells also expressed glucokinase, glucagon-like peptide 1 receptor (GLP1R), sulfonylurea receptor-1 (SUR1) and phogrin'all essential for glucose sensitivity and insulin secretion. The differentiated cells appropriately processed C-peptide and insulin in response to increasing glucose stimulation as shown by enzyme-linked immunosorbent assay (ELISA), fluorescence-activated cell sorting analysis, western blotting, and immunofluorescence staining. We provide a new approach using short-activating RNA in developing insulin producing surrogate cells for treating diabetes.http://www.sciencedirect.com/science/article/pii/S2162253116301561 |
| spellingShingle | Vikash Reebye Pål Sætrom Paul J Mintz John J Rossi Noriyuki Kasahara Georgios Nteliopoulos Joanna Nicholls Abdelali Haoudi Myrtle Gordon Nagy A Habib A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in CD34+ Cells Molecular Therapy: Nucleic Acids |
| title | A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in CD34+ Cells |
| title_full | A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in CD34+ Cells |
| title_fullStr | A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in CD34+ Cells |
| title_full_unstemmed | A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in CD34+ Cells |
| title_short | A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in CD34+ Cells |
| title_sort | short activating rna oligonucleotide targeting the islet β cell transcriptional factor mafa in cd34 cells |
| url | http://www.sciencedirect.com/science/article/pii/S2162253116301561 |
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