Reprogramming of Pancreatic Acinar Cells to Functional Beta Cells by In Vivo Transduction of a Polycistronic Construct Containing Pdx1, Ngn3, MafA in Mice.
To generate new beta cells after birth is a key focus of regenerative medicine, which could greatly aid the major health burden of diabetes. Beta-cell regeneration has been described using four different approaches: (1) the development of beta cells from putative precursor cells of the adult pancreas, which is termed neogenesis, (2) replication of existing beta cells, (3) differentiation from embryonic or induced pluripotent stem cells, and (4) reprogramming of non-beta cells to beta cells. Studies from the authors' laboratory have shown that beta-cell reprogramming can be achieved by transduction of adult pancreatic tissues with viral constructs containing the three developmentally important transcription factors Pdx1, Ngn3, and MafA. This protocol outlines the generation of a polycistronic construct containing the three transcription factors, the expansion and purification of the polycistronic virus, and in vivo transduction for acinar to beta-cell reprogramming in adult mice. The ultimate goal is to generate beta-like cells that resemble as closely as possible endogenous beta cells in phenotype and function for potential translational applications. © 2017 by John Wiley & Sons, Inc.
Section on Islet Cell and Regenerative Biology, Joslin Diabetes Center, Boston.|University Hospital of Basel, Department of Endocrinology, Diabetes, and Metabolism, Basel, Switzerland.
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Cavelti-Weder
Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge.|Covagen AG, Schlieren, Switzerland.
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Zumsteg
Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge.|Translational Medical Center for Stem Cell Therapy and Institute for Regenerative Medicine, Shanghai East Hospital, Tongji University School of Life Sciences and Technology, Shanghai.
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Li
Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge.
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Zhou
D061354:Acinar Cells / Q000378:metabolism*; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics*; D065150:Cellular Reprogramming / Q000235:genetics*; D018398:Homeodomain Proteins / Q000235:genetics*; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D051473:Maf Transcription Factors, Large / Q000235:genetics*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics*; D046790:Pancreas, Exocrine / Q000166:cytology*; D015534:Trans-Activators / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods*; D014780:Viruses / Q000378:metabolism
D051792:Basic Helix-Loop-Helix Proteins; D018398:Homeodomain Proteins; D051473:Maf Transcription Factors, Large; D009419:Nerve Tissue Proteins; D015534:Trans-Activators; C407737:Neurog3 protein, mouse; C084010:pancreatic and duodenal homeobox 1 protein