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

Research output: Contribution to journalArticle

8 Citations (Scopus)

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 - 2013
Externally publishedYes

Fingerprint

Islets of Langerhans
Oligonucleotides
RNA
Insulin
Sulfonylurea Receptors
Glucokinase
Glucose
Bioengineering
Fibrosarcoma
C-Peptide
Homeobox Genes
Insulin-Secreting Cells
Oncogenes
Fluorescent Antibody Technique
Insulin Resistance
Flow Cytometry
Western Blotting
Enzyme-Linked Immunosorbent Assay
Staining and Labeling
Genes

ASJC Scopus subject areas

  • Molecular Medicine
  • Drug Discovery

Cite this

A short-activating RNA oligonucleotide targeting the islet β-cell transcriptional factor MafA in CD34+ cells. / Reebye, Vikash; Sætrom, Pål; Mintz, Paul J.; Rossi, John J.; Kasahara, Noriyuki; Nteliopoulos, Georgios; Nicholls, Joanna; Haoudi, Abdelali; Gordon, Myrtle; Habib, Nagy A.

In: Molecular Therapy - Nucleic Acids, Vol. 2, e97, 2013.

Research output: Contribution to journalArticle

Reebye, V, Sætrom, P, Mintz, PJ, Rossi, JJ, Kasahara, N, Nteliopoulos, G, Nicholls, J, Haoudi, A, Gordon, M & Habib, NA 2013, 'A short-activating RNA oligonucleotide targeting the islet β-cell transcriptional factor MafA in CD34+ cells', Molecular Therapy - Nucleic Acids, vol. 2, e97. https://doi.org/10.1038/mtna.2013.23
Reebye, Vikash ; Sætrom, Pål ; Mintz, Paul J. ; Rossi, John J. ; Kasahara, Noriyuki ; Nteliopoulos, Georgios ; Nicholls, Joanna ; Haoudi, Abdelali ; Gordon, Myrtle ; Habib, Nagy A. / A short-activating RNA oligonucleotide targeting the islet β-cell transcriptional factor MafA in CD34+ cells. In: Molecular Therapy - Nucleic Acids. 2013 ; Vol. 2.
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