Adenovirus Transduction is Required for the Correction of Diabetes Using Pdx-1 or Neurogenin-3 in the Liver

Department of Molecular Pharmacology, School of Medicine, Stanford University, Stanford, California 94305, USA.
Molecular Therapy (Impact Factor: 6.23). 03/2007; 15(2):255-63. DOI: 10.1038/
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The regeneration of insulin-producing cells in vivo has emerged as a promising method for treating type I diabetes. Pancreatic duodenal homeobox-1 (Pdx-1), NeuroD, and Neurogenin-3 (Ngn3) are pancreatic transcription factors important for the development of insulin-producing cells in the liver. Other groups have demonstrated that adenoviral-mediated transgene expression of these transcription factors in the liver can reverse hyperglycemia in diabetic mice. We delivered Pdx-1 and Ngn3 to the livers of diabetic mice using adeno-associated virus (AAV) serotype 8, a vector that has been shown to result in non-toxic, persistent, high level expression of the transgene. We were unable to correct hyperglycemia in mice with streptozotocin-induced diabetes using AAV vectors expressing Pdx-1 and Ngn3. However, when we co-delivered these transcription factor expression cassettes in non-viral vectors with an irrelevant adenoviral vector, we were able to correct hyperglycemia in diabetic animals. Further studies demonstrated that an antigen-dependent immune response elicited by the adenoviral capsid together with the expression of a pancreatic transcription factor was required for restoration of serum insulin levels by the liver. Our results suggest that a host response to adenovirus in combination with expression of a pro-endocrine pancreas transcription factor is sufficient to induce insulin production in the livers of diabetic mice.

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Available from: Mark A Kay, Oct 06, 2015
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    • "Pdx1, MafA, and NeuroD/Beta2 induced expression of endogeneous insulin gene in liver [26]. Co-expression of Pdx1 and Ngn3 activated the transfected hepatocytes acquired the ability to synthesize and secrete insulin [24]. The miR-302$367 cluster encodes several miR-302 family members, including miR-302a/b/c/d and miR-367. "
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    ABSTRACT: The direct conversion of one cell type to another without an intermediate pluripotent stage is required for regenerative therapies. The ventral pancreas and liver share a common developmental origin. Recent studies have shown that hepatocytes could be induced to transdifferentiate into insulin-producing cells. In this paper, we showed a new strategy to achieve the direct conversion of human hepatocytes into surrogate β cells. Hepatocytes were transfected with microRNA-302 (miR-302) mimic and Pdx1, Ngn3 and MafA expressed plasmids, followed by a chemical-defined culture system for maturation of insulin-secreting cells. Co-transfection of miR-302 mimic increased the transcription of pancreatic development-related genes (Sox17, Foxa2, and endogenous Pdx1). Furthermore, at the end of this treatment, hepatocytes became insulin expressed cells that released the hormone in response to a physiological glucose change in vitro. This work shows that miR-302 participation may facilitates the conversion of adult hepatocytes into pancreatic islets-like cells.
    Biochemical and Biophysical Research Communications 09/2014; 453(3). DOI:10.1016/j.bbrc.2014.09.095 · 2.30 Impact Factor
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    • "Ferber et al. reported that ectopic expression of Pdx1 could induce the conversion from hepatocytes to IPCs and improve the hyperglycemia in streptozotocin- (STZ-) treated diabetic mouse [4]. Thereafter, several studies have confirmed that many other factors involved in pancreas development, including neurogenin 3 (Ngn3) [5], betacellulin [6], neurogenic differentiation (NeuroD) [7], and v-musculoaponeurotic fibrosarcoma oncogene homolog A (MafA) [8], could also turn on endocrine program in hepatocytes but not to generate functional beta-cells. Recently, an exciting and interesting breakthrough in islet regeneration was found by Melton and his colleagues. "
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    ABSTRACT: The neogenesis of insulin-producing cells (IPCs) from non-beta-cells has emerged as a potential method for treating diabetes mellitus (DM). Many groups have documented that activation of pancreatic transcription factor(s) in hepatocytes can improve the hyperglycemia in diabetic mice. In the present study, we explored a novel protocol that reprogrammed primary hepatocytes into functional IPCs by using multicistronic vectors carrying pancreatic and duodenal homeobox-1 (Pdx1), neurogenin 3 (Ngn3), and v-musculoaponeurotic fibrosarcoma oncogene homolog A (MafA). These triple-transfected cells activated multiple beta-cell genes, synthesized and stored considerable amounts of insulin, and released the hormone in a glucose-regulated manner in vitro. Furthermore, when transplanted into streptozotocin-induced diabetic mice, the cells markedly ameliorated glucose tolerance. Our results indicated that ectopic expression of Pdx1, Ngn3, and MafA facilitated hepatocytes-to-IPCs reprogramming. This approach may offer opportunities for treatment of DM.
    Journal of Diabetes Research 05/2014; 2014:716163. DOI:10.1155/2014/716163 · 2.16 Impact Factor
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    • "There are several potential explanations for the discrepancy between our results and previous studies reporting ''acinar reprogramming'' (Akinci et al., 2012, 2013; Zhou et al., 2008). First, the adenoviral delivery method employed by these studies may have established an inflammatory microenvironment that was more conducive to cellular transdifferentiation, as has been suggested (Lee et al., 2012; Wang et al., 2007). Alternatively, the in vivo conversion of acinar cells to b-like cells (Zhou et al., 2008) may have required a specific stoichiometry of PMN expression that was not achieved in the Tetß system. "
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    ABSTRACT: The ability to interconvert terminally differentiated cells could serve as a powerful tool for cell-based treatment of degenerative diseases, including diabetes mellitus. To determine which, if any, adult tissues are competent to activate an islet β cell program, we performed an in vivo screen by expressing three β cell "reprogramming factors" in a wide spectrum of tissues. We report that transient intestinal expression of these factors-Pdx1, MafA, and Ngn3 (PMN)-promotes rapid conversion of intestinal crypt cells into endocrine cells, which coalesce into "neoislets" below the crypt base. Neoislet cells express insulin and show ultrastructural features of β cells. Importantly, intestinal neoislets are glucose-responsive and able to ameliorate hyperglycemia in diabetic mice. Moreover, PMN expression in human intestinal "organoids" stimulates the conversion of intestinal epithelial cells into β-like cells. Our results thus demonstrate that the intestine is an accessible and abundant source of functional insulin-producing cells.
    Cell Reports 03/2014; 6(6). DOI:10.1016/j.celrep.2014.02.013 · 8.36 Impact Factor
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