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 language | English (US) |
|---|---|
| Article number | e97 |
| Journal | Molecular Therapy - Nucleic Acids |
| Volume | 2 |
| DOIs | |
| State | Published - 2013 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - A short-activating RNA oligonucleotide targeting the islet β-cell transcriptional factor MafA in CD34+ cells
AU - Reebye, Vikash
AU - Sætrom, Pål
AU - Mintz, Paul J.
AU - Rossi, John J.
AU - Kasahara, Noriyuki
AU - Nteliopoulos, Georgios
AU - Nicholls, Joanna
AU - Haoudi, Abdelali
AU - Gordon, Myrtle
AU - Habib, Nagy A.
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84878870592&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84878870592&partnerID=8YFLogxK
U2 - 10.1038/mtna.2013.23
DO - 10.1038/mtna.2013.23
M3 - Article
C2 - 23736775
AN - SCOPUS:84878870592
VL - 2
JO - Molecular Therapy - Nucleic Acids
JF - Molecular Therapy - Nucleic Acids
SN - 2162-2531
M1 - e97
ER -