title: Transduction of human and mouse pancreatic islet cells using a bicistronic recombinant adeno-associated viral vector.
abstract: Recent reports indicate successful transduction of pancreatic islets using recombinant adeno-associated viral (rAAV) vectors. This advance offers new possibilities in rendering islets resistant to rejection and recurrence of autoimmune destruction in the setting of islet transplantation as treatment of type 1 diabetes. Most gene delivery approaches using islets have thus far involved transduction with a single gene. However, the concomitant delivery of more than one gene encoding cytoprotective and/or immunoregulatory molecules may offer superior clinical utility. Here, we have generated a bicistronic rAAV (serotype 2) vector incorporating a viral internal ribosome entry site (IRES), derived from polio virus type 1, to allow for translation of two coupled cDNAs from a single mRNA transcript. Our study demonstrates the ability of this vector to produce significant expression of two reporter proteins in human and mouse islets in vitro. This expression did not interfere with beta-cell function. Transduction was maintained in vivo following transplantation of mouse islets. These data are the first report of efficient islet cell transduction with two genes using a single bicistronic rAAV vector and have direct implications for strategies aimed at enhancing islet transplant survival.
authors.affiliation: Department of Medicine, University of Florida, Gainesville, Florida 32610, USA.
authors.forename: Matthias
authors.initials: M
authors.lastname: Kapturczak
authors.forename: Sergei
authors.initials: S
authors.lastname: Zolotukhin
authors.forename: Jeff
authors.initials: J
authors.lastname: Cross
authors.forename: Antonello
authors.initials: A
authors.lastname: Pileggi
authors.forename: R Damaris
authors.initials: RD
authors.lastname: Molano
authors.forename: Marda
authors.initials: M
authors.lastname: Jorgensen
authors.forename: Barry
authors.initials: B
authors.lastname: Byrne
authors.forename: Terence R
authors.initials: TR
authors.lastname: Flotte
authors.forename: Tamir
authors.initials: T
authors.lastname: Ellis
authors.forename: Luca
authors.initials: L
authors.lastname: Inverardi
authors.forename: Camillo
authors.initials: C
authors.lastname: Ricordi
authors.forename: Harry
authors.initials: H
authors.lastname: Nick
authors.forename: Mark
authors.initials: M
authors.lastname: Atkinson
authors.forename: Anupam
authors.initials: A
authors.lastname: Agarwal
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology; D018014:Gene Transfer Techniques; D005796:Genes; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007515:Islets of Langerhans / Q000378:metabolism*; D051379:Mice; D014161:Transduction, Genetic