Evaluation of insulin expression and secretion in genetically engineered gut K and L-cells

Zalinah Ahmad,Mina Rasouli,Abdul Rahman Omar,Ahmad Zaid Fattah Azman

doi:10.1186/1472-6750-12-64

Abstract

BackgroundGene therapy could provide an effective treatment of diabetes. Previous studies have investigated the potential for several cell and tissue types to produce mature and active insulin. Gut K and L-cells could be potential candidate hosts for gene therapy because of their special features.ResultsIn this study, we isolated gut K and L-cells to compare the potential of both cell types to produce insulin when exposed to similar conditions. The isolated pure K and L-cells were transfected with recombinant plasmids encoding insulin and with specific promoters for K or L-cells. Insulin expression was studied in response to glucose or meat hydrolysate. We found that glucose and meat hydrolysate efficiently induced insulin secretion from K and L-cells. However, the effects of meat hydrolysate on insulin secretion were more potent in both cells compared with glucose. Results of enzyme-linked immunosorbent assays showed that L-cells secreted more insulin compared with K-cells regardless of the stimulator, although this difference was not statistically significant.ConclusionThe responses of K and L-cells to stimulation with glucose or meat hydrolysate were generally comparable. Therefore, both K and L-cells show similar potential to be used as surrogate cells for insulin gene expression in vitro. The potential use of these cells for diabetic gene therapy warrants further investigation.

Highlights

Gene therapy could provide an effective treatment of diabetes
We found that meat hydrolysate is a potent stimulator of insulin expression in both types
Isolation of stable intestinal K and L-cells The STC-1 cell line is a heterogeneous population of intestinal cells that was first isolated and characterized by Rindi et al 1990

Summary

Introduction

Gene therapy could provide an effective treatment of diabetes. Several approaches have been considered to develop effective and convenient treatments for diabetes, such as gene therapy [1,2,3]. Several gene therapy strategies have been developed to regulate physiologic blood glucose levels in patients with diabetes. One option that has been studied is to replace pancreatic β-cells with engineered nonpancreatic β-cells to generate potential target cells for gene therapy. These surrogate β-cells were able to synthesize and secrete active insulin; insulin secretion was not regulated by glucose, unlike endogenous

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