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Efficient gene delivery to human and rodent islets with double-stranded (ds) AAV-based vectors
K K Rehman1, Z Wang, R Bottino
1Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Gene Therapy
|April 29, 2005
Summary
Double-stranded adeno-associated viral (dsAAV) vectors efficiently gene-modify pancreatic islets for type 1 diabetes treatment. This approach enhances islet function and survival post-transplantation, bypassing the need for immunosuppression drugs.
Area of Science:
- Regenerative Medicine
- Gene Therapy
- Immunology
Background:
- Pancreatic islet transplantation is a viable treatment for type 1 diabetes.
- Systemic immunosuppression is required after transplantation, posing significant risks.
- Current gene transfer methods using adeno-associated viral (AAV) vectors have low transduction efficiency in islets.
Purpose of the Study:
- To evaluate the efficiency and functionality of double-stranded AAV (dsAAV) vectors for gene transfer into pancreatic islets.
- To assess the therapeutic potential of dsAAV-transduced islets in a type 1 diabetes model.
- To determine the immunogenicity and long-term expression of dsAAV-mediated transgenes in islet grafts.
Main Methods:
- Transduction of human and murine islets using single-stranded AAV2 (ssAAV2) and double-stranded AAV2 (dsAAV2) vectors expressing enhanced green fluorescent protein (eGFP).
- Assessment of islet function, viability, and insulin content post-transduction.
- Transplantation of dsAAV2-eGFP transduced islets into diabetic mice and monitoring of glycemic control and immune response.
- Evaluation of dsAAV serotype tropism for human islets.
Main Results:
- dsAAV2 vectors demonstrated significantly higher transduction efficiency in islets compared to ssAAV2 vectors.
- Murine islets transduced with dsAAV2-eGFP maintained normal glucose responsiveness, viability, and insulin content.
- Transplantation of dsAAV2-eGFP transduced islets restored normoglycemia in diabetic mice without immune rejection.
- Sustained eGFP expression was observed in grafts for at least 6 months post-transplantation.
- dsAAV serotypes 2, 6, and 8 efficiently transduced human islets.
Conclusions:
- Double-stranded AAV vectors offer a highly efficient platform for gene transfer into pancreatic islets.
- dsAAV-mediated gene modification of islets holds significant promise for improving islet transplantation outcomes in type 1 diabetes.
- This strategy may circumvent the need for systemic immunosuppression, enhancing patient safety and graft survival.