Abstract

INTRODUCTION. Type 1 diabetes is an autoimmune disease resulting in the destruction of the insulin-producing beta cells of the islets of Langerhans. The transplantation of healthy insulin-producing islets into patients with type 1 diabetes has been pursued for decades as an important treatment modality for this disease. Reproducible success of this approach has been complicated by early loss of transplanted cells due to apoptosis. The clinical utility of allogeneic islet transplantation will be tremendously enhanced if transplanted islets can be genetically engineered to evade pro-apoptotic stimuli at the engraftment site. Non-viral-based vectors such as lipofectants are good candidates to assess the role played by ex-vivo gene transfer of anti-apoptotic genes into primary islets. The aim of this study was to lipofect the dominant negative mutant of FADD (FADD-DN) into murine pancreatic islets to assay the effect of this transgene on islet viability.

Highlights

  • Type 1 diabetes is an autoimmune disease resulting in the destruction of the insulin-producing beta cells of the islets of Langerhans

  • Murine pancreatic islets were isolated and lipofected with plasmids coding for FADD DN (NFD-4)(1)

  • Barber) or the reporter Green Fluorescent Protein (GFP). Both these genes were cloned into a pLXSN-based plasmid (2) directly downstream of the LTR

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Summary

INTRODUCTION

Type 1 diabetes is an autoimmune disease resulting in the destruction of the insulin-producing beta cells of the islets of Langerhans. The transplantation of healthy insulin-producing islets into patients with type 1 diabetes has been pursued for decades as an important treatment modality for this disease. Reproducible success of this approach has been complicated by early loss of transplanted cells due to apoptosis. The clinical utility of allogeneic islet transplantation will be tremendously enhanced if transplanted islets can be genetically engineered to evade pro-apoptotic stimuli at the engraftment site. Non-viral-based vectors such as lipofectants are good candidates to assess the role played by ex-vivo gene transfer of anti-apoptotic genes into primary islets. The aim of this study was to lipofect the dominant negative mutant of FADD (FADD-DN) into murine pancreatic islets to assay the effect of this transgene on islet viability

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