Retrovirally transferred genes inhibit apoptosis in an insulin-secreting cell line: implications for islet transplantation.
The transplantation of pancreatic islets for the treatment of type I diabetes is hindered by the enormous loss of cells due to early apoptotic events. Genetic engineering of islets with cytoprotective genes is an important strategy aimed to enhance the survival of these cells in the transplant setting. The present study was designed to evaluate and compare the effects of five genes on a cell line derived from insulin-producing beta-cells, NIT-1. Cells were transduced using a Maloney murine leukemia virus (MLV) vector coding for yellow fluorescent protein (YFP) and for one of the following antiapoptotic genes: cFLIP, FADD-DN, BcL-2, PI-9, and ICAM-2. These genes were able to protect NIT-1 cells from cytokine-induced apoptosis to varying degrees ranging from no protection to significant protection equivalent to an optimal dose of a chemical caspase inhibitor. The data demonstrate that cFLIP, FADD-DN, and PI-9 are significantly more effective in protecting NIT-1 cells than BcL-2 and ICAM-2. Additionally, the data show that despite its weak in vitro inhibition of caspase-3, PI-9 affords significant protection against TNF-alpha-induced apoptosis in these cells. These genes may be ideal candidates to augment islet survival following transplantation.
Fenjves
Elizabeth S
ES
Diabetes Research Institute, University of Miami, School of Medicine, Miami, FL 33136, USA. Efenjves@miami.edu
Ochoa
M Sofia
MS
Gay-Rabinstein
Carlota
C
Ricordi
Camillo
C
Curran
Michael A
MA
D000818:Animals; D017209:Apoptosis; D001426:Bacterial Proteins / Q000235:genetics; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival; D017690:Cell Transplantation; D016207:Cytokines / Q000378:metabolism; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D005822:Genetic Vectors; D020287:In Situ Nick-End Labeling; D007328:Insulin / Q000378:metabolism*; D007340:Insulinoma / Q000378:metabolism; D016381:Islets of Langerhans Transplantation; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008954:Models, Biological; D008979:Moloney murine leukemia virus / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D013997:Time Factors; D019076:Transgenes; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D001426:Bacterial Proteins; D016207:Cytokines; D007328:Insulin; D008164:Luminescent Proteins; D014409:Tumor Necrosis Factor-alpha; C054562:yellow fluorescent protein, Bacteria; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases