title: Protection of human pancreatic islets using a lentiviral vector expressing two genes: cFLIP and GFP.
abstract: Pancreatic islet transplantation can provide insulin independence to diabetic patients. However, apoptosis of islets often leads to early graft failure. Genetic engineering with protective gene(s) can improve the viability of these cells. Here we show successful transduction of human islets with a feline immunodeficiency virus (FIV) vector expressing both a cytoprotective (cFLIP) gene and the green fluorescent protein (GFP). Despite using low virus titers to maximize safety, transduced islets expressed both genes, resulting in improved beta-cell metabolic activity and viability. Although only approximately 10% of total islet cells were transduced, the significant viability advantages suggest a "barrier" effect in which protecting the periphery of the islet shields the core. These results provide the first demonstration that a lentiviral vector can express two genes in islets. Furthermore, the engineered islets are resistant to a variety of apoptotic stimuli, suggesting the potential of this approach in enhancing the viability of transplanted cells.
authors.affiliation: Diabetes Research Institute, University of Miami School of Medicine, Miami, FL, USA.
authors.forename: Elizabeth S
authors.initials: ES
authors.lastname: Fenjves
authors.forename: M Sofia
authors.initials: MS
authors.lastname: Ochoa
authors.forename: Sirlene
authors.initials: S
authors.lastname: Cechin
authors.forename: Carlota
authors.initials: C
authors.lastname: Gay-Rabinstein
authors.forename: Ingrid
authors.initials: I
authors.lastname: Pérez-Alvarez
authors.forename: Hirohito
authors.initials: H
authors.lastname: Ichii
authors.forename: Armando
authors.initials: A
authors.lastname: Mendez
authors.forename: Camillo
authors.initials: C
authors.lastname: Ricordi
authors.forename: Michael A
authors.initials: MA
authors.lastname: Curran
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000378:metabolism; D017209:Apoptosis / Q000502:physiology; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein / Q000235:genetics* / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D004317:Doxorubicin / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006084:Graft Rejection / Q000517:prevention & control*; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D016089:Immunodeficiency Virus, Feline / Q000235:genetics* / Q000378:metabolism; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000502:physiology*; D019014:fas Receptor / Q000378:metabolism
chemical_list: D000903:Antibiotics, Antineoplastic; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; D016207:Cytokines; D007328:Insulin; D019014:fas Receptor; D049452:Green Fluorescent Proteins; D004317:Doxorubicin; D005947:Glucose