dsAAV8-mediated gene transfer and β-cell expression of IL-4 and β-cell growth factors are capable of reversing early-onset diabetes in NOD mice

Abstract

Type-I diabetes is a chronic disease mediated by autoimmune destruction of insulin-producing b-cells. Although progress has been made towards improving diabetes-associated pathologies and the quality of life for those living with diabetes, no therapy has been effective at eliminating disease manifestations or reversing disease progression. Here, we examined whether doublestranded adeno-associated virus serotype 8 (dsAAV8)-mediated gene delivery to endogenous b-cells of interleukin (IL)-4 in combination with b-cell growth factors can reverse early-onset diabetes in NOD mice. Our results demonstrate that a single treatment with dsAAV8 vectors expressing IL-4 in combination with glucagon-like peptide-1 or hepatocyte growth factor/NK1 under the regulation of the insulin promoter enhanced b-cell proliferation and survival in vivo, significantly delaying diabetes progression in NOD mice, and reversing disease in B10% of treated NOD mice. These results demonstrate the ability to reverse hyperglycemia in NOD mice with established diabetes by in vivo gene transfer to b-cells of immunomodulatory factors and b-cell growth factors.

Preparation of Preproinsulin Gene Construct Containing the Metallothionein2A (pBINDMTChIns) and Its Expression in NIH3T3 Cell Line and Muscle Tissue of Alloxan Diabetic Rabbits

“…Although, some developments have been made in improving pathologies of diabetes and the quality of patients' life, no treatment has been effective in reducing clinical complications or reversing progression of the disease (23). Until now, many studies have been carried out about type 1 diabetes gene therapy, but none of them have been done on larger animals like rabbits.…”

“…This is not perfect; however, it looks to be efficient (28). Many efforts have been made to prevent graft rejection such as constructing retroviral vector and double-stranded adeno-associated virus serotype 8 (dsAAV8) containing IL-4 gene in combination with β cell growth factors by Gaddy and Kapturczak (23,29). The Kapturczak`s designed vector had a desirable rate of protein expression, but there were some restrictions in carrying out and biosafety due to vector expression rates (29).…”

Transgene and islet cell delivery systems using nano-sized carriers for the treatment of diabetes mellitus

“…Protected against the development of diabetes mellitusRiedel et al (2010)[19] Double-stranded AAV-8GLP-1 and IL-4-DNAIntestinesIntraperitoneal injectionIncreased β cell mass and prevented β cell apoptosis. Decreased blood glucose levelGaddy et al (2012)[20] AAV-9GLP-1 and REG3 proteinIntestinesIntraperitoneal injectionPrevented hyperglycemia. Increased insulin-positive cell massTonne et al (2013)[21] AAV-8IL-2PancreasIntraperitoneal injectionPrevented onset of diabetesFlores et al (2014)[22] AAV-2KlothoPancreasIntraperitoneal injectionImproved glucose tolerance and attenuated β cell apoptosis.Lin and Sun (2015)[23]Enhanced insulin storage in β cells and increased plasma insulin levelsNon-viral vectorsPEIAFT3-siRNAPancreasTail vein injectionAttenuated ER stress-mediated pancreatic β cell dysfunctionKim et al (2013)[24] PEIFas-siRNAPancreasTail vein injectionDelayed the development of diabetes mellitusJeong et al (2010)[25] PEIGLP-1(7–37)-plasmid DNAIntestinesTail vein injectionIncreased insulin secretion.…”

“…Delivery of GLP-1 transgene with cationic nanomicelles comprising chitosan or arginine-grafted bioreducible polymer to the intestines was reported to have insulinotropic effects in animal models of diabetes mellitus [24,28,54]. It was reported that co-delivery of GLP-1 transgene to the intestines with other molecules could enhance the treatment effects for diabetes mellitus [18,20,21,26]. Co-delivery of GLP-1 transgene with N and K1 domains of hepatocyte growth factor (NK1/HGF) with double-stranded AAV vectors to the intestines was shown to delay the onset of diabetes mellitus, increase the mass of pancreatic β cells, and increase insulin secretion in an animal model of diabetes mellitus [18].…”

“…Co-delivery of GLP-1 transgene with N and K1 domains of hepatocyte growth factor (NK1/HGF) with double-stranded AAV vectors to the intestines was shown to delay the onset of diabetes mellitus, increase the mass of pancreatic β cells, and increase insulin secretion in an animal model of diabetes mellitus [18]. Other studies have demonstrated that co-delivery of GLP-1 transgene with interleukin (IL)-4 or NK1/HGF with double-stranded AAV-8 vectors to the intestines promoted pancreatic β cell proliferation and prevented pancreatic β cell apoptosis in an animal model of diabetes mellitus [20]. Co-delivery of GLP-1 transgene with regenerating islet-derived protein 3, which contributes to pancreatic islet neogenesis, with AAV vectors to the intestines was shown to decrease fasting blood glucose levels and increase the number of insulin-positive cells in the pancreas of a mouse model of diabetes mellitus [21].…”