A short-activating RNA oligonucleotide targeting the islet β-cell transcriptional factor MafA in CD34+ cells

Vikash Reebye, Pål Sætrom, Paul J. Mintz, John J. Rossi, Noriyuki Kasahara, Georgios Nteliopoulos, Joanna Nicholls, Abdelali Haoudi, Myrtle Gordon, Nagy A. Habib

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Abstract

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.

Original languageEnglish (US)
Article numbere97
JournalMolecular Therapy - Nucleic Acids
Volume2
DOIs
StatePublished - Jun 17 2013

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ASJC Scopus subject areas

  • Molecular Medicine
  • Drug Discovery

Cite this

Reebye, V., Sætrom, P., Mintz, P. J., Rossi, J. J., Kasahara, N., Nteliopoulos, G., Nicholls, J., Haoudi, A., Gordon, M., & Habib, N. A. (2013). A short-activating RNA oligonucleotide targeting the islet β-cell transcriptional factor MafA in CD34+ cells. Molecular Therapy - Nucleic Acids, 2, [e97]. https://doi.org/10.1038/mtna.2013.23