A cure for diabetes has long been sought using several different approaches, including islet transplantation, regeneration of β cells and insulin gene therapy1. However, permanent remission of type 1 diabetes has not yet been satisfactorily achieved. The development of type 1 diabetes results from the almost total destruction of insulin-producing pancreatic β cells by autoimmune responses specific to β cells2,3,4,5,6. Standard insulin therapy may not maintain blood glucose concentrations within the relatively narrow range that occurs in the presence of normal pancreatic β cells7. We used a recombinant adeno-associated virus (rAAV) that expresses a single-chain insulin analogue (SIA), which possesses biologically active insulin activity without enzymatic conversion, under the control of hepatocyte-specific L-type pyruvate kinase (LPK) promoter, which regulates SIA expression in response to blood glucose levels. Here we show that SIA produced from the gene construct rAAV-LPK-SIA caused remission of diabetes in streptozotocin-induced diabetic rats and autoimmune diabetic mice for a prolonged time without any apparent side effects. This new SIA gene therapy may have potential therapeutic value for the cure of autoimmune diabetes in humans.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Levine, F. & Leibowitz, G. Towards gene therapy of diabetes mellitus. Mol. Med. Today 5, 165–171 (1999).
Yoon, J. W. & Jun, H. S. in Encyclopedia of Immunology (eds Roitt, I. M. & Delves, P. J.) 2nd edn, 1390–1398 (Academic, London, 1998).
Schranz, D. B. & Lernmark, A. Immunology in diabetes: an update. Diab. Metab. Rev. 14, 3–29 (1998).
Tisch, R. & McDevitt, H. Insulin-dependent diabetes mellitus. Cell 85, 291–297 (1996).
Bach, J. F. Insulin-dependent diabetes mellitus as a β cell targeted disease of immunoregulation. J. Autoimmun. 8, 439–463 (1995).
Rossini, A. A., Greiner, D. L., Friedman, H. P. & Mordes, J. P. Immunopathogenesis of diabetes mellitus. Diabetes Rev. 1, 43–75 (1993).
The Diabetes Control and Complications Trial Research Group. The effect of intensive treatment of diabetes on the development and progression of long-term complications in insulin-dependent diabetes mellitus. New Engl. J. Med. 329, 977–986 (1993).
Cuif, M. H., Doiron, B. & Kahn, A. Insulin and cyclic AMP act at different levels on transcription of the L-type pyruvate kinase gene. FEBS Lett. 417, 81–84 (1997).
Chen, R., Doiron, B. & Kahn, A. Glucose responsiveness of a reporter gene transduced into hepatocytic cells using a retroviral vector. FEBS Lett. 365, 223–226 (1995).
Decaux, J. F., Antoine, B. & Kahn, A. Regulation of the expression of the L-type pyruvate kinase gene in adult rat hepatocytes in primary culture. J. Biol. Chem. 264, 11584–11590 (1989).
Cuif, M. H., Porteu, A., Kahn, A. & Vaulont, S. Exploration of a liver-specific, glucose/insulin-responsive promoter in transgenic mice. J. Biol. Chem. 268, 13769–13772 (1993).
Bergot, M. O., Diaz-Guerra, M. J., Puzenat, N., Raymondjean, M. & Kahn, A. Cis-regulation of the L-type pyruvate kinase gene promoter by glucose, insulin and cyclic AMP. Nucleic Acids Res. 20, 1871–1877 (1992).
Muzyczka, N. Use of adeno-associated virus as a general transduction vector for mammalian cells. Curr. Top. Microbiol. Immunol. 158, 97–129 (1992).
Clark, K. R., Liu, X., McGrath, J. P. & Johnson, P. R. Highly purified recombinant adeno-associated virus vectors are biologically active and free of detectable helper and wild-type viruses. Hum. Gene Ther. 10, 1031–1039 (1999).
Samulski, R. J. Adeno-associated virus: integration at a specific chromosomal locus. Curr. Opin. Genet. Dev. 3, 74–80 (1993).
Giraud, C., Winocour, E. & Berns, K. I. Site-specific integration by adeno-associated virus is directed by a cellular DNA sequence. Proc. Natl Acad. Sci. USA 91, 10039–10043 (1994).
Kotin, R. M., Linden, R. M. & Berns, K. I. Characterization of a preferred site on human chromosome 19q for integration of adeno-associated virus DNA by non-homologous recombination. EMBO J. 11, 5071–5078 (1992).
Cameron, N. E., Cotter, M. A. & Low, P. A. Nerve blood flow in early experimental diabetes in rats: relation to conduction deficits. Am. J. Physiol. 261, E1–E8 (1991).
Studier, F. W., Rosenberg, A. H., Dunn, J. J. & Dubendorff, J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 185, 60–89 (1990).
Pollet, R. J., Standaert, M. L. & Haase, B. A. Insulin binding to the human lymphocyte receptor. Evaluation of the negative cooperativity model. J. Biol. Chem. 252, 5828–5834 (1977).
Roth, J. Assay of peptide hormones using cell receptors: application to insulin and to human growth hormone. Methods Enzymol. 37, 66–82 (1975).
Frost, S. C. & Lane, M. D. Evidence for the involvement of vicinal sulfhydryl groups in insulin-activated hexose transport by 3T3-L1 adipocytes. J. Biol. Chem. 260, 2646–2652 (1985).
Heath, W. F. et al. (A-C-B) human proinsulin, a novel insulin agonist and intermediate in the synthesis of biosynthetic human insulin. J. Biol. Chem. 267, 419–425 (1992).
Yoon, J. W., Lesniak, M. A., Fussganger, R. & Notkins, A. L. Genetic differences in susceptibility of pancreatic β-cells to virus-induced diabetes mellitus. Nature 264, 178–180 (1976).
Ma, Z. et al. Effect of hemoglobin- and Perflubron-based oxygen carriers on common clinical laboratory tests. Clin. Chem. 43, 1732–1737 (1997).
Yoon, J. W., Rodrigues, M. M., Currier, C. & Notkins, A. Long-term complications of virus-induced diabetes mellitus in mice. Nature 296, 566–569 (1982).
Yoon, J. W. et al. Control of autoimmune diabetes in NOD mice by GAD expression or suppression in cells. Science 284, 1183–1187 (1999).
Hirasawa, K. Possible role of macrophage-derived soluble mediators in the pathogenesis of EMC virus-induced diabetes in mice. J. Virol. 71, 4024–4031 (1997).
We thank R. J. Samulski for providing psub201 and pXX2, A. L. Kyle and K. Clarke for editorial assistance and B. Pinder for artwork. We also thank H. S. Jun for a critical review of the manuscript. J. W. Y. is a Heritage Medical Scientist awardee of the Alberta Heritage Foundation for Medical Research. This work was supported in part by Brain Korea 21 Project.
About this article
Toxicology Mechanisms and Methods (2019)
Advanced Healthcare Materials (2019)
Misfolded proinsulin impairs processing of precursor of insulin receptor and insulin signaling in β cells
The FASEB Journal (2019)
The Korean Journal of Urological Oncology (2017)
mRNA destabilization improves glycemic responsiveness of transcriptionally regulated hepatic insulin gene therapyin vitroandin vivo
The Journal of Gene Medicine (2017)