A short-activating RNA oligonucleotide targeting the islet beta-cell transcriptional factor MafA in CD34(+) cells
Author(s)
Type
Journal Article
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.
Date Issued
2016-12-14
Online Publication Date
2016-12-14
2017-05-15T12:03:47Z
Date Acceptance
2013-03-27
ISSN
2162-2531
Publisher
NATURE PUBLISHING GROUP
Journal / Book Title
MOLECULAR THERAPY-NUCLEIC ACIDS
Volume
2
Copyright Statement
© 2013 American Society of Gene & Cell Therapy. Published by Elsevier Inc. Molecular Therapy–Nucleic Acids
is an open-access
journal published by
Nature Publishing Group
. This work
is licensed under a Creative Commons Attribution-NonCommercial-
NoDerivative Works 3.0 License. To view a copy of this license, visit
http://creativecommons.org/licenses/by-nc-nd/3.0/
is an open-access
journal published by
Nature Publishing Group
. This work
is licensed under a Creative Commons Attribution-NonCommercial-
NoDerivative Works 3.0 License. To view a copy of this license, visit
http://creativecommons.org/licenses/by-nc-nd/3.0/
Source Database
web-of-science
Identifier
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000332466500002&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
Subjects
Science & Technology
Life Sciences & Biomedicine
Medicine, Research & Experimental
Research & Experimental Medicine
EMBRYONIC STEM-CELLS
INSULIN-PRODUCING CELLS
IN-VITRO
PROMOTE DIFFERENTIATION
ANTISENSE TRANSCRIPTS
CLINICAL-APPLICATION
MAMMALIAN-CELLS
ENDOCRINE-CELLS
GENE-EXPRESSION
BONE-MARROW
Publication Status
Published
Article Number
ARTN e97