Developing a Sensitive Reporter System for Monitoring of Pancreatic and Duodenal Homeobox Gene 1 (Pdx1) and Neurogenin 3 (Ngn3) – Mediated Transdifferentiation from Human Hepatic Cells into Insulin-Producing Beta-Like Cells

Abstract:

It is well known that cellular differentiation is not a terminal process. Transdifferentiation is the conversion of one differentiated cell type to another. There are many examples of induced transdifferentiation between cell types by expression of ectopic transcription factors. Here we show that combined lentiviral expression of Pdx1 or Pdx1-VP16 fusion protein and Ngn3 can direct the transdifferentiation of hepatic cells into insulin producing cells. We showed that the Pdx1 or Pdx1-VP16 fusion protein and Ngn3 together synergistically increased transactivation for the insulin gene. This provides a useful model to study the transdifferentiation process.

• Periodical: Advanced Materials Research (Volumes 989-994)
• Pages: 1003-1006
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.989-994.1003
• Online since: July 2014
• Authors: Chao Xie*, William Donelan, Shun Lu, Li Jun Yang
• Keywords: Insulin, Ngn3, Pdx1, Transdifferentiation

References

[1] Slack J.M. & Tosh,D. Transdifferentiation and metaplasia-switching cell types. Curr. Opin. Genet. Dev. 11, 581-586 (2001).

DOI: 10.1016/s0959-437x(00)00236-7

[2] Eguchi,G. & Kodama,R. Transdifferentiation. Curr. Opin. Cell Biol. 5, 1023-1028 (1993).

[3] Li,W.C., Yu,W.Y., Quinlan J.M., Burke Z.D. & Tosh,D. The molecular basis of transdifferentiation. J. Cell Mol. Med. 9, 569-582 (2005).

DOI: 10.1111/j.1582-4934.2005.tb00489.x

[4] Weintraub,H. et al. Activation of muscle-specific genes in pigment, nerve, fat, liver, and fibroblast cell lines by forced expression of MyoD. Proc. Natl. Acad. Sci. U. S. A 86, 5434-5438 (1989).

DOI: 10.1073/pnas.86.14.5434

[5] Horb M.E., Shen C.N., Tosh,D. & Slack J.M. Experimental conversion of liver to pancreas. Curr. Biol. 13, 105-115 (2003).

DOI: 10.1016/s0960-9822(02)01434-3

[6] Watada,H. Neurogenin 3 is a key transcription factor for differentiation of the endocrine pancreas. Endocr. J. 51, 255-264 (2004).

DOI: 10.1507/endocrj.51.255

[7] Cao L.Z., Tang D.Q., Horb M.E., Li,S.W. & Yang L.J. High glucose is necessary for complete maturation of Pdx1-VP16-expressing hepatic cells into functional insulin-producing cells. Diabetes 53, 3168-3178 (2004).

DOI: 10.2337/diabetes.53.12.3168

[8] Tang D.Q. et al. In vivo and in vitro characterization of insulin-producing cells obtained from murine bone marrow. Diabetes 53, 1721-1732 (2004).

DOI: 10.2337/diabetes.53.7.1721

[9] Tang D.Q. et al. Reprogramming liver-stem WB cells into functional insulin-producing cells by persistent expression of Pdx1- and Pdx1-VP16 mediated by lentiviral vectors. Lab Invest 86, 83-93 (2006).

DOI: 10.1038/labinvest.3700368

[10] Chang L.J. & Zaiss A.K. Lentiviral vectors. Preparation and use. Methods Mol. Med. 69, 303-318 (2002).

[11] Blouin,A., Bolender R.P. & Weibel E.R. Distribution of organelles and membranes between hepatocytes and nonhepatocytes in the rat liver parenchyma. A stereological study. J. Cell Biol. 72, 441-455 (1977).

DOI: 10.1083/jcb.72.2.441

[12] Zhao,R. & Duncan S.A. Embryonic development of the liver. Hepatology 41, 956-967 (2005).

[13] Derman,E. et al. Transcriptional control in the production of liver-specific mRNAs. Cell 23, 731-739 (1981).

DOI: 10.1016/0092-8674(81)90436-0