Inhibition of ZEB1 expression induces redifferentiation of adult human [beta] cells expanded in vitro

In-vitro expansion of functional adult human β-cells is an attractive approach for generating insulin-producing cells for transplantation. However, human islet cell expansion in culture results in loss of β-cell phenotype and epithelial-mesenchymal transition (EMT). This process activates expression...

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Bibliographic Details
Published inScientific reports Vol. 5; p. 13024
Main Authors Sintov, Elad, Nathan, Gili, Knoller, Sarah, Pasmanik-chor, Metsada, Russ, Holger A, Efrat, Shimon
Format Journal Article
LanguageEnglish
Published London Nature Publishing Group 01.08.2015
Subjects
Beta cells
Cell culture
Cell proliferation
Diabetes mellitus
E-cadherin
Gene expression
Glucose
Homeobox
Insulin
Insulin secretion
Islet cells
Kinases
Mesenchyme
Pancreatic islet transplantation
Repressors
Secretion
Transplantation
Zinc finger proteins
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Summary:In-vitro expansion of functional adult human β-cells is an attractive approach for generating insulin-producing cells for transplantation. However, human islet cell expansion in culture results in loss of β-cell phenotype and epithelial-mesenchymal transition (EMT). This process activates expression of ZEB1 and ZEB2, two members of the zinc-finger homeobox family of E-cadherin repressors, which play key roles in EMT. Downregulation of ZEB1 using shRNA in expanded β-cell-derived (BCD) cells induced mesenchymal-epithelial transition (MET), β-cell gene expression, and proliferation attenuation. In addition, inhibition of ZEB1 expression potentiated redifferentiation induced by a combination of soluble factors, as judged by an improved response to glucose stimulation and a 3-fold increase in the fraction of C-peptide-positive cells to 60% of BCD cells. Furthermore, ZEB1 shRNA led to increased insulin secretion in cells transplanted in vivo. Our findings suggest that the effects of ZEB1 inhibition are mediated by attenuation of the miR-200c target genes SOX6 and SOX2. These findings, which were reproducible in cells derived from multiple human donors, emphasize the key role of ZEB1 in EMT in cultured BCD cells and support the value of ZEB1 inhibition for BCD cell redifferentiation and generation of functional human β-like cells for cell therapy of diabetes.
ISSN:2045-2322
DOI:10.1038/srep13024