title: Evaluation of promoters for driving efficient transgene expression in neonatal porcine islets.
abstract: There is considerable interest in the viral modification of insulin-producing islets, including porcine islets, in the context of islet xenotransplantation to treat type 1 diabetes. Adenovirus (Adv) gene delivery offers the potential to modify pre-transplant islets for enhanced survival. Modifications include transfer of cytoprotective molecules to ensure islet survival immediately post-transplant, and molecules to dampen the immune system and prevent chronic islet graft rejection. In this study, we compared different promoters (three promiscuous and two tissue-specific promoters) for their efficiency in driving gene expression in neonatal pig islet tissue after Adv delivery. We also compared the efficiency of these promoters in adult islets from mouse and human pancreata. We observed that the promiscuous cytomegalovirus promoter was the most potent, eliciting high luciferase expression in neonatal pig islets, as well as in human and mouse islets. In contrast, the mammalian EF1-alpha promoter educed comparatively intermediate gene expression. The mouse major histocompatibility complex class I promoter H-2K(b) and the pancreatic-specific promoters insulin and human pdx-1 (area II) performed poorly in islets from all three species. This has important implications for the generation of modified neonatal pig islets for transplantation into humans.
authors: The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia.
authors: Sarah L
authors: SL
authors: Londrigan
authors: Jamie L
authors: JL
authors: Brady
authors: Robyn M
authors: RM
authors: Sutherland
authors: Wayne J
authors: WJ
authors: Hawthorne
authors: Helen E
authors: HE
authors: Thomas
authors: Guarang
authors: G
authors: Jhala
authors: Peter J
authors: PJ
authors: Cowan
authors: Thomas W H
authors: TW
authors: Kay
authors: Philip J
authors: PJ
authors: O'Connell
authors: Andrew M
authors: AM
authors: Lew
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D006183:H-2 Antigens / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D016381:Islets of Langerhans Transplantation / Q000379:methods; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D013552:Swine; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D019076:Transgenes / Q000235:genetics / Q000502:physiology*; D014183:Transplantation, Heterologous / Q000379:methods
chemical_list: D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D018398:Homeodomain Proteins; D007328:Insulin; D020648:Peptide Elongation Factor 1; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D008156:Luciferases