title: A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in CD34(+) Cells.
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.Molecular Therapy - Nucleic Acids (2013) 2, e97; doi:10.1038/mtna.2013.23; advance online publication 4 June 2013.
authors.affiliation: Department of Surgery and Cancer, Imperial College London, London, UK.
authors.forename: Vikash
authors.initials: V
authors.lastname: Reebye
authors.forename: Pål
authors.initials: P
authors.lastname: Sætrom
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Mintz
authors.forename: John J
authors.initials: JJ
authors.lastname: Rossi
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kasahara
authors.forename: Georgios
authors.initials: G
authors.lastname: Nteliopoulos
authors.forename: Joanna
authors.initials: J
authors.lastname: Nicholls
authors.forename: Abdelali
authors.initials: A
authors.lastname: Haoudi
authors.forename: Myrtle
authors.initials: M
authors.lastname: Gordon
authors.forename: Nagy A
authors.initials: NA
authors.lastname: Habib