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A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in CD34+ Cells

journal contribution
submitted on 2024-07-23, 09:53 and posted on 2024-07-23, 09:53 authored by Vikash Reebye, Pål Sætrom, Paul J Mintz, John J Rossi, Noriyuki Kasahara, Georgios Nteliopoulos, Joanna Nicholls, Abdelali Haoudi, Myrtle Gordon, Nagy A Habib

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.

Other Information

Published in: Molecular Therapy - Nucleic Acids
License: https://creativecommons.org/licenses/by-nc-nd/3.0/
See article on publisher's website: https://dx.doi.org/10.1038/mtna.2013.23

History

Language

  • English

Publisher

Cell Press

Publication Year

  • 2013

License statement

This Item is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 International License.

Institution affiliated with

  • Hamad Bin Khalifa University
  • Qatar Biomedical Research Institute - HBKU