Insulin expressing hepatocytes not destroyed in transgenic NOD mice

Tabiin, Muhammad T; White, Curt M.; Morahan, Grant; Tuch, Bernard E.

doi:10.1186/1740-2557-1-3

Cited by 21 publications

(9 citation statements)

References 24 publications

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“…Hemotoxylin and eosin stained sections of livers from diabetic rats, harvested after 4 days and up to a month after TA1m treatment did not show any indication of T-cell infiltration or the presence of anti-insulin antibody in the serum of TA1m treated rats. Our results are consistent with previously published studies reporting that insulin expression in liver does not cause the hepatocyte to become a target of immune attack [46].…”

Section: Discussionsupporting

confidence: 94%

Correction of Diabetic Hyperglycemia and Amelioration of Metabolic Anomalies by Minicircle DNA Mediated Glucose-Dependent Hepatic Insulin Production

et al. 2013

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Type 1 diabetes mellitus (T1DM) is caused by immune destruction of insulin-producing pancreatic β-cells. Commonly used insulin injection therapy does not provide a dynamic blood glucose control to prevent long-term systemic T1DM-associated damages. Donor shortage and the limited long-term success of islet transplants have stimulated the development of novel therapies for T1DM. Gene therapy-based glucose-regulated hepatic insulin production is a promising strategy to treat T1DM. We have developed gene constructs which cause glucose-concentration–dependent human insulin production in liver cells. A novel set of human insulin expression constructs containing a combination of elements to improve gene transcription, mRNA processing, and translation efficiency were generated as minicircle DNA preparations that lack bacterial and viral DNA. Hepatocytes transduced with the new constructs, ex vivo, produced large amounts of glucose-inducible human insulin. In vivo, insulin minicircle DNA (TA1m) treated streptozotocin (STZ)-diabetic rats demonstrated euglycemia when fasted or fed, ad libitum. Weight loss due to uncontrolled hyperglycemia was reversed in insulin gene treated diabetic rats to normal rate of weight gain, lasting ∼1 month. Intraperitoneal glucose tolerance test (IPGT) demonstrated in vivo glucose-responsive changes in insulin levels to correct hyperglycemia within 45 minutes. A single TA1m treatment raised serum albumin levels in diabetic rats to normal and significantly reduced hypertriglyceridemia and hypercholesterolemia. Elevated serum levels of aspartate transaminase, alanine aminotransferase, and alkaline phosphatase were restored to normal or greatly reduced in treated rats, indicating normalization of liver function. Non-viral insulin minicircle DNA-based TA1m mediated glucose-dependent insulin production in liver may represent a safe and promising approach to treat T1DM.

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Section: Discussionsupporting

confidence: 94%

Correction of Diabetic Hyperglycemia and Amelioration of Metabolic Anomalies by Minicircle DNA Mediated Glucose-Dependent Hepatic Insulin Production

et al. 2013

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“…Second, hepatocytes are of non-beta cell types that do not succumb to autoimmune attack. Hepatic insulin gene transfer has the potential of restoring endogenous insulin production for long-term glycemic control without eliciting recurrent autoimmunity against insulin-producing hepatocytes [32] . However, achieving adequately regulated hepatic insulin production in coupling with blood glucose remains a major hurdle, although progress has been made to regulate insulin production in the liver, using the L-PK promoter [50–52] .…”

Section: Discussionmentioning

confidence: 99%

“…As an alternative approach, the liver is being sought as a surrogate organ for restoring endogenous insulin production. Unlike transplanted islets that succumb to autoimmune destruction, insulin-producing hepatocytes are of non-β-cell type and are refractory to autoimmunity [32] . Similar to β-cells, hepatocytes possess the glucose-sensing mechanism due to the expression of glucokinase (GK) and glucose transporter 2 (Glut2) [33] .…”

Section: Introductionmentioning

confidence: 99%

Glucose-regulated insulin production in the liver improves glycemic control in type 1 diabetic mice

Zhang

Dong

2015

Molecular Metabolism

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ObjectiveType 1 diabetes results from autoimmune destruction of beta-cells in the pancreas. Our objective is to reconstitute a glucose-responsive system in the liver to regulate hepatic insulin production for improving glycemic control in type 1 diabetes.MethodsWe have cloned the glucose-responsive element (GRE) from the promoter of acetyl-CoA carboxylase (ACC), an enzyme that catalyzes the rate-limiting step in fatty acid synthesis in the liver in response to glucose. To increase the amplitude of glucose induction, we quadruplicated the GRE DNA by gene duplication. The resulting GRE multimer (4×GRE) was tested for its ability to drive rat proinsulin cDNA expression in hepatocytes and insulin-deficient diabetic mice.ResultsWe showed that this GRE multimer-directed glucose-responsive system produced insulin in hepatocytes in a glucose-dependent manner. When delivered into the liver by adenovirus-mediated gene transfer, this glucose-responsive insulin production system was able to reverse hyperglycemia to a normal range without causing hypoglycemia after glucose challenge or overnight fasting. Insulin vector-treated diabetic mice exhibited significantly improved blood glucose profiles in response to glucose tolerance, correlating with insulin production in the liver. We recapitulated these findings in streptozotocin-induced diabetic CD1 mice and autoimmune non-obese diabetic mice.ConclusionOur data characterized the GRE motif from the ACC promoter as a potent glucose-responsive element, and provided proof-of-concept that the 4×GRE-mediated hepatic insulin production is capable of correcting insulin deficiency and improving glycemic control in type 1 diabetes.

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“…This implies that the ‘neo-islets’ induced by Ngn3-Btc appear to behave, immunologically, in a very similar fashion as mature native or transplanted islets in that they trigger an identical autoimmune response that targets pancreatic islets. This, of in itself, is very different from earlier gene therapy studies that had attempted in a reversal of diabetes in NOD mice by either producing insulin in the hepatocytes (NOD-PEPCK-Ins Tg) 22 or converting the hepatocytes into insulin producing cells using PDX1 gene therapy 23 , as in both of these studies the insulin producing cells behave differently as compared to native mature β-cells and appear not to be targeted indicating that these were not recognized as β-cells by the immune system.…”

Section: Discussionmentioning

confidence: 66%

Gene therapy with neurogenin3, betacellulin and SOCS1 reverses diabetes in NOD mice

Buras

Lee

et al. 2015

Gene Ther

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Islet transplantation for Type 1 diabetes is limited by a shortage of donor islets and requirement for immunosuppression. We approached this problem by inducing in vivo islet neogenesis in NOD diabetic mice, a model of autoimmune diabetes. We demonstrate that gene therapy with helper-dependent adenovirus (HDAd) carrying neurogenin3, an islet lineage-defining transcription factor and betacellulin, an islet growth factor, leads to the induction of periportal insulin-positive cell clusters in the liver, which are rapidly destroyed. To specifically accord protection to these ‘neo-islets’ from cytokine-mediated destruction, we overexpressed suppressor of cytokine signaling 1 (SOCS1) gene, using a rat insulin promoter in combination with neurogenin3 and betacellulin. With this approach, about half of diabetic mice attained euglycemia sustained for over 4 months, regain glucose tolerance and appropriate glucose-stimulated insulin secretion. Histological analysis revealed periportal islet hormone-expressing ‘neo-islets’ in treated mouse livers. Despite evidence of persistent ‘insulitis’ with activated T-cells, these ‘neo-islets’ persist to maintain euglycemia. This therapy does not affect diabetogenicity of splenocytes, as they retain the ability to transfer diabetes. This study thus provides a proof-of-concept for engineering in vivo islet neogenesis with targeted resistance to cytokine-mediated destruction to provide a long-term reversal of diabetes in NOD mice.

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Insulin expressing hepatocytes not destroyed in transgenic NOD mice

Cited by 21 publications

References 24 publications

Correction of Diabetic Hyperglycemia and Amelioration of Metabolic Anomalies by Minicircle DNA Mediated Glucose-Dependent Hepatic Insulin Production

Correction of Diabetic Hyperglycemia and Amelioration of Metabolic Anomalies by Minicircle DNA Mediated Glucose-Dependent Hepatic Insulin Production

Glucose-regulated insulin production in the liver improves glycemic control in type 1 diabetic mice

Gene therapy with neurogenin3, betacellulin and SOCS1 reverses diabetes in NOD mice

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