Widespread and stable pancreatic gene transfer by adeno-associated virus vectors via different routes.
Diabetes is a disease of epidemic proportions and is on the rise worldwide. Gene therapy has been actively pursued but limited by technical hurdles and profound inefficiency of direct gene transfer to the pancreas in vivo. Here, we show that, for the first time, appropriate serotypes of adeno-associated virus (AAV), coupled with a double-stranded vector DNA cassette, enable extensive and long-term in vivo gene transfer in the adult mouse pancreas by three different delivery methods. Intraperitoneal and intravenous delivery of AAV8 effectively transduced exocrine acinar cells as well as endocrine beta-cells, while local pancreatic intraductal delivery of AAV6 showed the best efficiency in the beta-cells among all AAV serotypes tested in this study. Nearly the entire islet population showed gene transfer but with distinct gene transfer efficiency and patterns when different delivery methods and vectors were used. Importantly, localized gene delivery coupled with an insulin promoter allowed extensive yet specific gene expression in the beta-cells. These effective new methods should provide useful tools to study diabetes pathogenesis and gene therapy.
Department of Orthopedic Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Zhong
Z
Wang
Tong
T
Zhu
Khaja K
KK
Rehman
Suzanne
S
Bertera
Jian
J
Zhang
Chunlian
C
Chen
Glenn
G
Papworth
Simon
S
Watkins
Massimo
M
Trucco
Paul D
PD
Robbins
Juan
J
Li
Xiao
X
Xiao
D000199:Actins / Q000235:genetics; D000818:Animals; D002645:Chickens; D017931:DNA Primers; D000229:Dependovirus; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D007515:Islets of Langerhans / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D010179:Pancreas / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000032:analysis
D000199:Actins; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins