Evaluation of promoters for driving efficient transgene expression in neonatal porcine islets.
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.
Londrigan
Sarah L
SL
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia.
Brady
Jamie L
JL
Sutherland
Robyn M
RM
Hawthorne
Wayne J
WJ
Thomas
Helen E
HE
Jhala
Guarang
G
Cowan
Peter J
PJ
Kay
Thomas W H
TW
O'Connell
Philip J
PJ
Lew
Andrew M
AM
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
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