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Retrovirally transferred genes inhibit apoptosis in an insulin-secreting cell line: implications for islet
Elizabeth S Fenjves1, M Sofia Ochoa, Carlota Gay-Rabinstein
1Diabetes Research Institute, University of Miami, School of Medicine, Miami, FL 33136, USA. Efenjves@miami.edu
Abstract:
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.
Insights
Genetic engineering enhances pancreatic islet cell survival for type I diabetes treatment. Genes like cFLIP, FADD-DN, and PI-9 show significant protection against apoptosis, improving transplant outcomes.
Area of Science:
- Immunology
- Cell Biology
- Genetics
Background:
- Type I diabetes treatment faces challenges with pancreatic islet cell loss due to apoptosis.
- Genetic engineering offers a strategy to improve islet cell survival post-transplantation.
- Identifying effective cytoprotective genes is crucial for successful islet transplantation.
Purpose of the Study:
- To evaluate and compare the efficacy of five anti-apoptotic genes (cFLIP, FADD-DN, BcL-2, PI-9, ICAM-2) in protecting insulin-producing beta-cells.
- To assess the protective effects against cytokine-induced apoptosis in a NIT-1 cell line.
- To determine the potential of these genes for augmenting islet survival in transplantation.
Main Methods:
- Utilized a Maloney murine leukemia virus (MLV) vector for gene transduction.
- Transduced NIT-1 cells (insulin-producing beta-cell line) with yellow fluorescent protein (YFP) and one of five anti-apoptotic genes.
- Assessed protection against cytokine-induced apoptosis and in vitro caspase-3 inhibition.
Main Results:
- All five tested genes demonstrated varying degrees of protection against cytokine-induced apoptosis.
- cFLIP, FADD-DN, and PI-9 provided significantly greater protection compared to BcL-2 and ICAM-2.
- PI-9 offered substantial protection against TNF-alpha-induced apoptosis despite weak in vitro caspase-3 inhibition.
Conclusions:
- cFLIP, FADD-DN, and PI-9 are effective candidates for enhancing pancreatic islet cell survival.
- These genes show promise for improving the success of islet transplantation in type I diabetes.
- Further investigation into PI-9's mechanism of action is warranted.
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