Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Human insulin production and amelioration of diabetes in mice by electrotransfer-enhanced plasmid DNA gene transfer to the skeletal muscle

Gene Therapy volume 9, pages 1429–1437 (2002)Cite this article

Abstract

A first-line gene therapy for type 1 diabetes should be based on a safe procedure to engineer an accessible tissue for insulin release. We evaluated the ability of the skeletal muscle to release human insulin after electrotransfer (ET)-enhanced plasmid DNA injection in mice. A furin-cleavable proinsulin cDNA under the CMV or the MFG promoter was electrotransferred to immune-incompetent mice with STZ-induced severe diabetes. At 1 week, mature human insulin was detected in the serum of 17/20 mice. After an initial peak of 68.5 ± 34.9 μU/ml, insulin was consistently detected at significant levels up to 6 weeks after gene transfer. Importantly, untreated diabetic animals died within 3 weeks after STZ, whereas treated mice survived up to 10 weeks. Fed blood glucose (BG) was reduced in correspondence with the insulin peak. Fasting BG was near-normalized when insulin levels were 12.9 ± 5.3 (CMV group, 2 weeks) and 7.7 ± 2.6 μU/ml (MFG group, 4 weeks), without frank hypoglycemia. These data indicate that ET-enhanced DNA injection in muscle leads to the release of biologically active insulin, with restoration of basal insulin levels, and lowering of fasting BG with increased survival in severe diabetes. Therefore the skeletal muscle can be considered as a platform for basal insulin secretion.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Diabetes Control and Complication Trial Research Group (DCCT). The effect of intensive treatment on the development and progression of long-term complications in insulin-dependent diabetes mellitus N Engl J Med 1993 329: 977–986

  2. Gautier JF et al. Are the implications of the Diabetes Control and Complications Trial (DCCT) feasible in daily clinical practice? Diabetes Metab 1996 22: 415–419

    CAS  PubMed  Google Scholar 

  3. Diabetes Control and Complication Trial Research Group (DCCT). Hypoglycemia in the Diabetes Control and Complications Trial Diabetes 1997 46: 271–286

  4. Hanaire-Broutin H et al. Comparison of continuous subcutaneous insulin infusion and multiple daily injection regimens using insulin Lispro in type 1 diabetic patients on intensified treatment Diabetes Care 2000 23: 1232–1235

    Article  CAS  Google Scholar 

  5. Scavini M et al. Catheter survival during long-term insulin therapy with an implanted programmable pump. The Implantable Insulin Pump Trial Study group Diabetes Care 1997 20: 610–613

    Article  CAS  Google Scholar 

  6. Thompson JS, Duckworth WC, Saudek CD, Giobbie-Hurder A . Surgical experience with implatable insulin pumps. Department of Veterans Affairs Implantable Insulin Pump Study Group Am J Surg 1998 176: 622–626

    Article  CAS  Google Scholar 

  7. Selam JL . External and implantable insulin pumps: current place in the treatment of diabetes Exp Clin Endocrinol Diabetes 2001 109: S333–S340

    Article  CAS  Google Scholar 

  8. Selam JL . External and implantable insulin pumps: current place in the treatment of diabetes Gruessner AC., Sutherland DE. Analysis of pancreas transplant outcomes for United States cases reported to the United Network for Organ Sharing (UNOS) and non-US cases reported to the International Pancreas Transplant Registry (IPTR). Clin Transpl 1999; 51-69.

  9. Martinenghi S et al. Cancer arising after pancreas and/or kidney transplantation in a series of 99 diabetic patients Diabetes Care 1997 20: 272–275

    Article  CAS  Google Scholar 

  10. Lumbreras C et al. Infectious complications following pancreatic transplantation: incidence, microbiological and clinical characteristics, and outcome Clin Infect Dis 1995 20: 514–520

    Article  CAS  Google Scholar 

  11. Smets YF et al. Infectious disease complications of simultaneous pancreas kidney transplantation Nephrol Dial Transpl 1997 12: 764–771

    Article  CAS  Google Scholar 

  12. Secchi A et al. Islet transplantation in IDDM patients Diabetologia 1997 40: 225–231

    Article  CAS  Google Scholar 

  13. Shapiro AM et al. Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen N Engl J Med 2000 343: 230–238

    Article  CAS  Google Scholar 

  14. Onkamo P, Vaananen S, Karvonen M, Tuomilehto J . Worldwide increase in incidence of type I diabetes - the analysis of the data on published incidence trends Diabetologia 1999 42: 1395–1403

    Article  CAS  Google Scholar 

  15. Karvonen M et al. Incidence of childhood type 1 diabetes worldwide. Diabetes Mondiale (DiaMond) Project Group Diabetes Care 2000 23: 1516–1526

    Article  CAS  Google Scholar 

  16. Song S et al. Sustained secretion of human alpha-1-antitrypsin from murine muscle transduced with adeno-associated virus vectors Proc Natl Acad Sci USA 1998 95: 14394–14398

    Google Scholar 

  17. Haase G et al. Adenovirus-mediated transfer of the neurotrophin-3 gene into skeletal muscle of pmn mice: therapeutic effects and mechanisms of action J Neurol Sci 1998 160: S97–S105

    Article  CAS  Google Scholar 

  18. Kay MA et al. Evidence for gene transfer and expression of factor IX in haemophilia B patients treated with an AAV vector Nat Genetics 2000 24: 257–261

    Article  CAS  Google Scholar 

  19. Tripathy SK et al. Long-term expression of erythropoietin in the systemic circulation of mice after intramuscular injection of a plasmid DNA vector Proc Natl Acad Sci USA 1996 93: 10876–10880

    Article  CAS  Google Scholar 

  20. Kon OL et al. Naked plasmid-mediated gene transfer to skeletal muscle ameliorates diabetes mellitus J Gene Med 1999 1: 186–194

    Article  CAS  Google Scholar 

  21. Athanasopoulos T et al. Intramuscular injection of a plasmid vector expressing human apolipoprotein E limits progression of xanthoma and aortic atheroma in apoE-deficient mice Hum Mol Genet 2000 9: 2545–2551

    Article  CAS  Google Scholar 

  22. Wolff JA et al. Direct gene transfer into mouse muscle in vivo Science 1990 247: 1465–1468

    Article  CAS  Google Scholar 

  23. Wolff JA et al. Conditions affecting direct gene transfer into rodent muscle in vivo Biotechniques 1991 11: 474–485

    CAS  Google Scholar 

  24. Wolff JA et al. Expression of naked plasmids by cultured myotubes and entry of plasmids into T tubules and caveolae of mammalian skeletal muscle J Cell Sci 1992 103: 1249–1259

    CAS  Google Scholar 

  25. Aihara H, Miyazaki JI . Gene transfer into muscle by electroporation in vivo Nat Biotechnol 1998 16: 867–870

    Article  CAS  Google Scholar 

  26. Mir LM et al. High-efficiency gene transfer into skeletal muscle mediated by electric pulses Proc Natl Acad Sci USA 1999 96: 4262–4267

    Article  CAS  Google Scholar 

  27. Draghia-Akli R et al. Myogenic expression of an injectable protease-resistant growth hormone releasing hormone augments long-term growth in pigs Nat Biotechnol 1999 17: 1179–1183

    Article  CAS  Google Scholar 

  28. Maruyama H et al. Continuous erythropoietin delivery by muscle-targeted gene transfer using in vivo electroporation Hum Gene Ther 2000 11: 429–437

    Article  CAS  Google Scholar 

  29. Rizzuto G et al. Gene electrotransfer results in a high level transduction of rat skeletal muscle and correct anemia of renal failure Hum Gene Ther 2000 11: 1891–1900

    Article  CAS  Google Scholar 

  30. Groskreutz DJ, Sliwkowski MX, Gorman CM . Genetically engineered proinsulin constitutively processed and secreted as mature, active insulin J Biol Chem 1994 269: 6241–6245

    CAS  Google Scholar 

  31. Falqui L et al. Reversal of diabetes in mice by implantation of human fibroblasts genetically engineered to release mature human insulin Hum Gene Ther 1999 10: 1753–1760

    Article  CAS  Google Scholar 

  32. Dong H et al. Basal insulin gene expression significantly improves conventional insulin therapy in type 1 diabetic rats Diabetes 2002 51: 130–138

    Article  CAS  Google Scholar 

  33. Lee HC et al. Remission in models of type 1 diabetes by gene therapy using a single-chain insulin analogue Nature 2000 408: 483–488

    Article  CAS  Google Scholar 

  34. Yin D, Tang J-G . Gene therapy for streptozotocin-induced diabetic mice by electroporational transfer of naked human insulin precursor DNA into skeletal muscle in vivo FEBS Lett 2001 495: 16–20

    Article  CAS  Google Scholar 

  35. Shifrin AL et al. Adenoviral vector-mediated insulin gene transfer in the mouse pancreas corrects streptozotocin-induced hyperglycemia Gene Therapy 2001 8: 1480–1489

    Article  CAS  Google Scholar 

  36. Cheung AT et al. Glucose-dependent insulin release from genetically engineered K cells Science 2000 290: 1959–1962

    Article  CAS  Google Scholar 

  37. Simonson GD, Grokreutz DJ, Gorman CM, Macdonald MJ . Synthesis and processing of genetically modified human proinsulin by rat myoblast primary cultures Hum Gene Ther 1996 7: 71–78

    Article  CAS  Google Scholar 

  38. Arcelloni C et al. Processing and release of human proinsulin-cleavage products into culture media by different engineered non-endocrine cells: a specific assessment by capillary electrophoresis J Endocrinol 2000 166: 437–445

    Article  CAS  Google Scholar 

  39. McMahon JM et al. Optimisation of electrotransfer of plasmid into skeletal muscle by pretreatment with hyaluronidase - increased expression with reduced muscle damage Gene Ther 2001 8: 1264–1270

    Article  CAS  Google Scholar 

  40. Hartikka J et al. Electroporation-facilitated delivery of plasmid DNA in skeletal muscle: plasmid dependence of muscle damage and effect of poloxamer 188 Mol Ther 2001 4: 407–415

    Article  CAS  Google Scholar 

  41. Skarli M et al. Myosin regulatory elements as vectors for gene transfer by intramuscular injection Gene Therapy 1998 5: 514–520

    Article  CAS  Google Scholar 

  42. Li X, Eastman EM, Schwartz RJ, Draghia-Akli R . Synthetic muscle promoters: activities exceeding naturally occurring regulatory sequences Nat Biotechnol 1999 17: 241–245

    Article  CAS  Google Scholar 

  43. Abai AM, Hobart PM, Barnhart KM . Insulin delivery with plasmid DNA Hum Gene Ther 1999 10: 2637–2649

    Article  CAS  Google Scholar 

  44. Stewart T, Taylor NA, Docherty K, Bailey CJ . Insulin delivery by somatic cell gene therapy J Mol Endocrinol 1993 11: 335–341

    Article  CAS  Google Scholar 

  45. Diabetes Control and Complication Trial Research Group (DCCT). Effects of age, duration and treatment of insulin-dependent diabetes mellitus on residual beta-cell function: observations during eligibility testing for the diabetes control and complication trial (DCCT) J Clin Endocrinol Metab 1987 65: 30–36

  46. Dranoff G et al. Vaccination with irradiated tumor cells engineered to secrete murine granulocyte-macrophage colony-stimulating factor stimulates potent, specific, and long-lasting anti-tumor immunity Proc Natl Acad Sci USA 1993 90: 3539–3543

    Article  CAS  Google Scholar 

  47. Manthorpe M et al. Gene therapy by intramuscular injection of plasmid DNA: studies on firefly luciferase gene expression in mice Hum Gene Ther 1993 4: 419–431

    Article  CAS  Google Scholar 

  48. Ferrari G et al. Muscle regeneration by bone marrow-derived myogenic progenitors Science 1998 279: 1528–1530

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy

    S Martinenghi, S Biressi, F Bifari, M G Roncarolo, C Bordignon & L Falqui

  2. Department of Medicine, Scientific Institute H. San Raffaele, Milan, Italy

    S Martinenghi & L Falqui

  3. Department of Neurology, Scientific Institute H San Raffaele, Milan, Italy

    S Amadio

  4. Department of Experimental Medicine, University of Pavia, Pavia, Italy

    G Cusella De Angelis

Authors
  1. S Martinenghi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G Cusella De Angelis
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S Biressi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S Amadio
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. F Bifari
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M G Roncarolo
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. C Bordignon
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. L Falqui
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Martinenghi, S., Cusella De Angelis, G., Biressi, S. et al. Human insulin production and amelioration of diabetes in mice by electrotransfer-enhanced plasmid DNA gene transfer to the skeletal muscle. Gene Ther 9, 1429–1437 (2002). https://doi.org/10.1038/sj.gt.3301804

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301804

Keywords

This article is cited by

Search

Advanced search

Quick links