Abstract
Due to the limitations of current treatment regimes, gene therapy is a promising strategy being explored to correct blood glucose concentrations in diabetic patients. In the current study, we used a retroviral vector to deliver either the human insulin gene alone, the rat NeuroD1 gene alone, or the human insulin gene and rat NeuroD1 genes together, to the rat liver cell line, H4IIE, to determine if storage of insulin and pancreatic transdifferentiation occurred. Stable clones were selected and expanded into cell lines: H4IIEins (insulin gene alone), H4IIE/ND (NeuroD1 gene alone), and H4IIEins/ND (insulin and NeuroD1 genes). The H4IIEins cells did not store insulin; however, H4IIE/ND and H4IIEins/ND cells stored 65.5 ± 5.6 and 1475.4 ± 171.8 pmol/insulin/5 × 106 cells, respectively. Additionally, several β cell transcription factors and pancreatic hormones were expressed in both H4IIE/ND and H4IIEins/ND cells. Electron microscopy revealed insulin storage vesicles in the H4IIE/ND and H4IIEins/ND cell lines. Regulated secretion of insulin to glucose (0–20 mmol/L) was seen in the H4IIEins/ND cell line. The H4IIEins/ND cells were transplanted into diabetic immunoincompetent mice, resulting in normalization of blood glucose. This data shows that the expression of NeuroD1 and insulin in liver cells may be a useful strategy for inducing islet neogenesis and reversing diabetes.
Highlights
Diabetes mellitus is a chronic metabolic disorder of multiple etiologies [1,2]
Diabetes is classified into three categories: type 1 diabetes mellitus (T1D), type 2 diabetes mellitus (T2D), and gestational diabetes [1]
By comparison H4IIE/ND and H4IIEins/ND cells stored and secreted both human and rat insulin following transduction, H4IIEins/ND cells stored and secreted significantly higher amounts of both human and rat insulin compared to other cell lines
Summary
The condition is characterized by chronic hyperglycaemia caused by immune-mediated beta (β) cell elimination or defects in insulin secretion and/or insulin action [1]. According to the International Diabetes Federation (IDF) [3], 415 million people have diabetes, and estimate that one person will die from diabetes-related conditions every six seconds. By 2040, it is estimated that 642 million people will have diabetes. T1D is caused by an absolute insulin deficiency, secondary to the autoimmune destruction of the insulin-secreting pancreatic β-cells [1]. T2D is characterized by insulin deficiency or insulin secretory defects, either with or without insulin resistance, and may be treated with oral medications, lifestyle, and dietary modifications; a large proportion of patients become insulin-dependent due to β-cell dysfunction. Gestational diabetes is characterized by hyperglycaemia caused by the inability to compensate for increased insulin demands during pregnancy [1]
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