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Engineering physiologically regulated insulin secretion in non-β cells by expressing glucagon-like peptide 1 receptor

Abstract

Glucagon-like peptide 1 (GLP-1) is released from neuroendocrine cells in the intestine in the postprandial state and augments glucose-stimulated insulin secretion from pancreatic β cells. To develop non-β cells that exhibit physiologically regulated insulin secretion, we coexpressed the GLP-1 receptor and human insulin in primary rat pituitary cells using adenovirus-mediated gene transfer. The transduced cells were analyzed in a perifusion system and after transplantation into mice. Normal pituitary cells do not express the GLP-1 receptor as shown by the absence of GLP-1 receptor mRNA and the inability of GLP-1 to stimulate pituitary hormone secretion. Following transduction with an adenovirus carrying the GLP-1 receptor cDNA, the pituitary cells expressed functional GLP-1 receptors as reflected by the ability of GLP-1 to stimulate secretion of pituitary hormones. When both the GLP-1 receptor and human insulin were introduced, GLP-1 stimulated cosecretion of human insulin and endogenous pituitary hormones. GLP-1 was similar in potency to the hypothalamic-releasing hormones and stimulated hormone secretion in a dose-dependent fashion. In contrast to pancreatic β cells, the hormone-releasing effect of GLP-1 on transduced pituitary cells was not dependent on the concentration of extracellular glucose. After transplantation of pituitary cells coexpressing human insulin and GLP-1 receptor into mice, enteral glucose stimulated insulin secretion. These results demonstrate a new approach to engineer physiologically regulated insulin secretion by non-β cells.

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References

  1. Perley MJ, Kipnis DM . Plasma insulin responses to oral and intravenous glucose: studies in normal and diabetic subjects. J Clin Invest 1967; 46: 1954–1962.

    Article  CAS  Google Scholar 

  2. Creutzfeldt M . Candidate hormones of the gut. XV. Insulin-releasing factors of the gastrointestinal mucosa (Incretin). Gastroenterology 1974; 67: 748–750.

    CAS  PubMed  Google Scholar 

  3. Holst JJ . Glucagon-like peptide 1 (GLP-1): an intestinal hormone, signalling nutritional abundance, with an unusual therapeutic Potential. Trends Endocrinol Metab 1999; 10: 229–235.

    Article  CAS  Google Scholar 

  4. Drucker DJ . Minireview: the glucagon-like peptides. Endocrinology 2001; 142: 521–527.

    Article  CAS  Google Scholar 

  5. Thorens B . Glucagon-like peptide-1 and control of insulin secretion. Diabetes Metab 1995; 21: 311–318.

    CAS  Google Scholar 

  6. Thorens B . Expression cloning of the pancreatic beta cell receptor for the gluco- incretin hormone glucagon-like peptide 1. Proc Natl Acad Sci USA 1992; 89: 8641–8645.

    Article  CAS  Google Scholar 

  7. Thorens B, Waeber G . Glucagon-like peptide-I and the control of insulin secretion in the normal state and in NIDDM. Diabetes 1993; 42: 1219–1225.

    Article  CAS  Google Scholar 

  8. Fehmann HC, Goke R, Goke B . Cell and molecular biology of the incretin hormones glucagon-like peptide-I and glucose-dependent insulin releasing polypeptide. Endocr Rev 1995; 16: 390–410.

    Article  CAS  Google Scholar 

  9. Sherwood NM, Krueckl SL, McRory JE . The origin and function of the pituitary adenylate cyclase-activating polypeptide (PACAP)/glucagon superfamily. Endocr Rev 2000; 21: 619–670.

    CAS  PubMed  Google Scholar 

  10. Holz GG et al. cAMP-dependent mobilization of intracellular Ca2+ stores by activation of ryanodine receptors in pancreatic beta-cells. A Ca2+ signaling system stimulated by the insulinotropic hormone glucagon-like peptide-1-(7-37). J Biol Chem 1999; 274: 14147–14156.

    Article  CAS  Google Scholar 

  11. Meglasson MD, Matschinsky FM . Pancreatic islet glucose metabolism and regulation of insulin secretion. Diabetes Metab Rev 1986; 2: 163–214.

    Article  CAS  Google Scholar 

  12. Fehmann HC et al. The effects of glucagon-like peptide-I (GLP-I) on hormone secretion from isolated human pancreatic islets. Pancreas 1995; 11: 196–200.

    Article  CAS  Google Scholar 

  13. Yamada S et al. Time-dependent stimulation of insulin exocytosis by 3′,5′-cyclic adenosine monophosphate in the rat islet beta-cell. Endocrinology 2002; 143: 4203–4209.

    Article  CAS  Google Scholar 

  14. Dachicourt N et al. Glucagon-like peptide-1(7-36)-amide confers glucose sensitivity to previously glucose-incompetent beta-cells in diabetic rats: in vivo and in vitro studies. J Endocrinol 1997; 155: 369–376.

    Article  CAS  Google Scholar 

  15. Gutniak M et al. Antidiabetogenic effect of glucagon-like peptide-1 (7-36)amide in normal subjects and patients with diabetes mellitus. N Engl J Med 1992; 326: 1316–1322.

    Article  CAS  Google Scholar 

  16. D'Alessio DA, Kahn SE, Leusner CR, Ensinck JW . Glucagon-like peptide 1 enhances glucose tolerance both by stimulation of insulin release and by increasing insulin-independent glucose disposal. J Clin Invest 1994; 93: 2263–2266.

    Article  CAS  Google Scholar 

  17. Ahren B, Larsson H, Holst JJ . Effects of glucagon-like peptide-1 on islet function and insulin sensitivity in noninsulin-dependent diabetes mellitus. J Clin Endocrinol Metab 1997; 82: 473–478.

    CAS  PubMed  Google Scholar 

  18. Nauck MA et al. Normalization of fasting hyperglycaemia by exogenous glucagon-like peptide 1 (7-36 amide) in type 2 (non-insulin-dependent) diabetic patients. Diabetologia 1993; 36: 741–744.

    Article  CAS  Google Scholar 

  19. Clark SA et al. Novel insulinoma cell lines produced by iterative engineering of GLUT2, glucokinase, and human insulin expression. Diabetes 1997; 46: 958–967.

    Article  CAS  Google Scholar 

  20. Hohmeier HE et al. Regulation of insulin secretion from novel engineered insulinoma cell lines. Diabetes 1997; 46: 968–977.

    Article  CAS  Google Scholar 

  21. Ferber S et al. Pancreatic and duodenal homeobox gene 1 induces expression of insulin genes in liver and ameliorates streptozotocin-induced hyperglycemia. Nat Med 2000; 6: 568–572.

    Article  CAS  Google Scholar 

  22. 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 

  23. Chen R, Meseck M, McEvoy RC, Woo SL . Glucose-stimulated and self-limiting insulin production by glucose 6-phosphatase promoter driven insulin expression in hepatoma cells. Gene Therapy 2000; 7: 1802–1809.

    Article  CAS  Google Scholar 

  24. Tooze SA . Biogenesis of secretory granules. Implications arising from the immature secretory granule in the regulated pathway of secretion. FEBS Lett 1991; 285: 220–224.

    Article  CAS  Google Scholar 

  25. Tooze SA, Martens GJ, Huttner WB . Secretory granule biogenesis: rafting to the SNARE. Trends Cell Biol 2001; 11: 116–122.

    Article  CAS  Google Scholar 

  26. Zhu X et al. Disruption of PC1/3 expression in mice causes dwarfism and multiple neuroendocrine peptide processing defects. Proc Natl Acad Sci USA 2002; 99: 10293–10298.

    Article  CAS  Google Scholar 

  27. Zhu X et al. Severe block in processing of proinsulin to insulin accompanied by elevation of des-64,65 proinsulin intermediates in islets of mice lacking prohormone convertase 1/3. Proc Natl Acad Sci USA 2002; 99: 10299–10304.

    Article  CAS  Google Scholar 

  28. Beak SA et al. Glucagon-like peptide-1 (GLP-1) releases thyrotropin (TSH): characterization of binding sites for GLP-1 on alpha-TSH cells. Endocrinology 1996; 137: 4130–4138.

    Article  CAS  Google Scholar 

  29. Taniguchi Y, Yasutaka S, Kominami R, Shinohara H . Proliferation and differentiation of rat anterior pituitary cells. Anat Embryol (Berl) 2002; 206: 1–11.

    Article  CAS  Google Scholar 

  30. Kendall SK et al. Targeted disruption of the pituitary glycoprotein hormone alpha-subunit produces hypogonadal and hypothyroid mice. Genes Dev 1995; 9: 2007–2019.

    Article  CAS  Google Scholar 

  31. Castro MG et al. Expression of transgenes in normal and neoplastic anterior pituitary cells using recombinant adenoviruses: long term expression, cell cycle dependency, and effects on hormone secretion. Endocrinology 1997; 138: 2184–2194.

    Article  CAS  Google Scholar 

  32. Holz GGt, Kuhtreiber WM, Habener JF . Pancreatic beta-cells are rendered glucose-competent by the insulinotropic hormone glucagon-like peptide-1(7-37). Nature 1993; 361: 362–365.

    Article  CAS  Google Scholar 

  33. Hui H, Yu R, Bousquet C, Perfetti R . Transfection of pancreatic-derived beta-cells with a minigene encoding for human glucagon-like peptide-1 regulates glucose-dependent insulin synthesis and secretion. Endocrinology 2002; 143: 3529–3539.

    Article  CAS  Google Scholar 

  34. Kolligs F, Fehmann HC, Goke R, Goke B . Reduction of the incretin effect in rats by the glucagon-like peptide 1 receptor antagonist exendin (9-39) amide. Diabetes 1995; 44: 16–19.

    Article  CAS  Google Scholar 

  35. Efrat S et al. Conditional transformation of a pancreatic beta-cell line derived from transgenic mice expressing a tetracycline-regulated oncogene. Proc Natl Acad Sci USA 1995; 92: 3576–3580.

    Article  CAS  Google Scholar 

  36. Lipes MA et al. Insulin-secreting non-islet cells are resistant to autoimmune destruction. Proc Natl Acad Sci USA 1996; 93: 8595–8600.

    Article  CAS  Google Scholar 

  37. Faradji RN et al. Glucose-induced toxicity in insulin-producing pituitary cells that coexpress GLUT2 and glucokinase. Implications for metabolic engineering. J Biol Chem 2001; 276: 36695–36702.

    Article  CAS  Google Scholar 

  38. Kolodka TM, Finegold M, Moss L, Woo SL . Gene therapy for diabetes mellitus in rats by hepatic expression of insulin. Proc Natl Acad Sci USA 1995; 92: 3293–3297.

    Article  CAS  Google Scholar 

  39. Gros L et al. Insulin production by engineered muscle cells. Hum Gene Ther 1999; 10: 1207–1217.

    Article  CAS  Google Scholar 

  40. 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: 1753–1762.

    Article  CAS  Google Scholar 

  41. Hughes SD, Johnson JH, Quaade C, Newgard CB . Engineering of glucose-stimulated insulin secretion and biosynthesis in non-islet cells. Proc Natl Acad Sci USA 1992; 89: 688–692.

    Article  CAS  Google Scholar 

  42. Hughes SD et al. Transfection of AtT-20ins cells with GLUT-2 but not GLUT-1 confers glucose-stimulated insulin secretion. Relationship to glucose metabolism. J Biol Chem 1993; 268: 15205–15012.

  43. Lu M, Wheeler MB, Leng XH, Boyd AEd . The role of the free cytosolic calcium level in beta-cell signal transduction by gastric inhibitory polypeptide and glucagon-like peptide I(7-37). Endocrinology 1993; 132: 94–100.

    Article  CAS  Google Scholar 

  44. Wei Y, Mojsov S . Tissue-specific expression of the human receptor for glucagon-like peptide-I: brain, heart and pancreatic forms have the same deduced amino acid sequences. FEBS Lett 1995; 358: 219–224.

    Article  CAS  Google Scholar 

  45. Wu L, Fritz JD, Powers AC . Different functional domains of GLUT2 glucose transporter are required for glucose affinity and substrate specificity. Endocrinology 1998; 139: 4205–4212.

    Article  CAS  Google Scholar 

  46. Watanabe T, Orth DN . Detailed kinetic analysis of adrenocorticotropin secretion by dispersed rat anterior pituitary cells in a microperifusion system: effects of ovine corticotropin-releasing factor and arginine vasopressin. Endocrinology 1987; 121: 1133–1145.

    Article  CAS  Google Scholar 

  47. Openshaw P et al. Heterogeneity of intracellular cytokine synthesis at the single-cell level in polarized T helper 1 and T helper 2 populations. J Exp Med 1995; 182: 1357–1367.

    Article  CAS  Google Scholar 

  48. Wang T et al. An encapsulation system for the immunoisolation of pancreatic islets. Nat Biotechnol 1997; 15: 358–362.

    Article  CAS  Google Scholar 

  49. Hartling SG et al. ELISA for human proinsulin. Clin Chim Acta 1986; 156: 289–297.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by a postdoctoral training grant from the Juvenile Diabetes Research Foundation International, a Merit Review Award from the VA Research Service, a research grant from the National Institutes of Health (DK55233), research grants from the American Diabetes Association and the Juvenile Diabetes Research Foundation International, and the Vanderbilt Diabetes Research and Training Center (NIH DK20593). The GLP-1 receptor cDNA was kindly provided by Dr Svetlana Mojsov at The Rockefeller University. The proinsulin assay was graciously performed at the Diabetes Research and Training Center at the University of Chicago (NIH DK20595) by Diane Ostrega and Dr Kenneth Polonsky.

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Correspondence to A C Powers.

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Wu, L., Nicholson, W., Wu, CY. et al. Engineering physiologically regulated insulin secretion in non-β cells by expressing glucagon-like peptide 1 receptor. Gene Ther 10, 1712–1720 (2003). https://doi.org/10.1038/sj.gt.3302055

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