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In vivo studies on non-viral transdifferentiation of liver cells towards pancreatic beta cells

Research output: Contribution to journalArticlepeer-review

Abdullah Cim, Greta J. Sawyer, Xiaohong Zhang, Haibin Su, Louise Collins, Peter Jones, Michael Antoniou, Jean-Paul Reynes, Hans-Joachim Lipps, John W. Fabre

Original languageEnglish
Pages (from-to)277-288
Number of pages12
JournalJournal of Endocrinology
Volume214
Issue number3
Early online date8 Jun 2012
DOIs
E-pub ahead of print8 Jun 2012
PublishedSep 2012

King's Authors

Abstract

Transdifferentiation in vivo is an attractive option for autologous replacement of pancreatic beta cells in patients with type 1 diabetes. It has been achieved by adenoviral delivery of genes for transcription factors in the liver and pancreas of hyperglycaemic mice. However, these viral approaches are not clinically applicable. We used the hydrodynamic approach to deliver genes Pdx1, Ngn3 (Neurog3) and MafA singly and in combination to livers of normoglycaemic rats. Five expression plasmids were evaluated. Livers were removed 1, 3, 7, 14 and 28 days after gene delivery and assayed by quantitative PCR, semi-quantitative PCR and immunohistology. Functional studies on hyperglycaemic rats were performed. The highest and most sustained expression was from a CpG-depleted plasmid (pCpG) and a plasmid with an in-frame scaffold/matrix attachment region ((pEPI(CMV)). When Pdx1, Ngn3 and MafA were delivered together to normoglycaemic rats with these plasmids, insulin mRNA was detected at all time points and was similar to 50-fold higher with pCpG. Insulin mRNA content of livers at days 3 and 7 was equivalent to that of a pancreas, with scattered insulin-positive cells detected by immunohistology, but levels declined thereafter. Prohormone convertase 1/3 was elevated at days 3 and 7. In hyperglycaemic rats, fasting blood glucose was lower at days 1, 3 and 7 but not thereafter, and body weight was maintained to day 28. We conclude that hydrodynamic gene delivery of multiple transcription factors to rat liver can initiate transdifferentiation to pancreatic beta cells, but the process is reversible and probably requires more sustained transcription factor expression. Journal of Endocrinology (2012) 214, 277-288

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