A stable analogue of glucose-dependent insulinotropic polypeptide, GIP(LysPAL16), enhances functional differentiation of mouse embryonic stem cells into cells expressing islet-specific genes and hormones
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Lamin Marenah
, Jane T. McCluskey , Yasser H.A. Abdel-Wahab , Finbarr P.M. O'Harte , Neville H. McClenaghan and Peter R. Flatt
Abstract
Embryonic stem (ES) cells can be differentiated into insulin-producing cells by conditioning the culture media. However, the number of insulin-expressing cells and amount of insulin released is very low. Glucose-dependent insulinotropic polypeptide (GIP) enhances the growth and differentiation of pancreatic β-cells. This study examined the potential of the stable analogue GIP(LysPAL16) to enhance the differentiation of mouse ES cells into insulin-producing cells using a five-stage culturing strategy. Semi-quantitative PCR indicated mRNA expression of islet development markers (nestin, Pdx1, Nkx6.1, Oct4), mature pancreatic β-cell markers (insulin, glucagon, Glut2, Sur1, Kir6.1) and the GIP receptor gene GIP-R in undifferentiated (stage 1) cells, with increasing levels in differentiated stages 4 and 5. IAPP and somatostatin genes were only expressed in differentiated stages. Immunohistochemical studies confirmed the presence of insulin, glucagon, somatostatin and IAPP in differentiated ES cells. After supplementation with GIP(LysPAL16), ES cells at stage 4 released insulin in response to secretagogues and glucose in a concentration-dependent manner, with 35–100% increases in insulin release. Cellular C-peptide content also increased by 45% at stages 4 and 5. We conclude that the stable GIP analogue enhanced differentiation of mouse ES cells towards a phenotype expressing specific β-cell genes and releasing insulin.
References
Abraham, E.J., Leech, C.A., Lin, J.C., Zulewski, H., and Habener, J.F. (2002). Insulinotropic hormone glucagon-like peptide-1 differentiation of human pancreatic islet-derived progenitor cells into insulin-producing cells. Endocrinology143, 3152–3161.10.1210/endo.143.8.8973Search in Google Scholar
Ahlgren, U., Jonsson, J., and Edlund, H. (1996). The morphogenesis of the pancreatic mesenchyme is uncoupled from that of the pancreatic epithelium in IPF1/PDX1-deficient mice. Development122, 1409–1416.10.1242/dev.122.5.1409Search in Google Scholar
Assady, S., Maor, G., Amit, M., Itskovitz-Eldor, J., Skorecki, K.L., and Tzukerman, M. (2001). Insulin production by human embryonic stem cells. Diabetes50, 1691–1697.10.2337/diabetes.50.8.1691Search in Google Scholar
Bai, L., Meredith, G., and Tuch, B.E. (2005). Glucagon-like peptide-1 enhances production of insulin-producing cells derived from mouse embryonic stem cells. J. Endocrinol.186, 343–352.10.1677/joe.1.06078Search in Google Scholar
Bonner-Weir, S., Taneja, M., Weir, G.C., Tatarkiewicz, K., Song, K.H., Sharma, A., and O'Neil, J.J. (2000). In vitro cultivation of human islets from expanded ductal tissue. Proc. Natl. Acad. Sci. USA97, 7999–8004.10.1073/pnas.97.14.7999Search in Google Scholar
Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem.72, 248–254.10.1016/0003-2697(76)90527-3Search in Google Scholar
Doetschman, T.C., Eistetter, H., Katz, M., Schmidt, W., and Kemler, R. (1985). The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. Embryol. Exp. Morphol.87, 27–45.10.1242/dev.87.1.27Search in Google Scholar
Drucker, D.J. (2002). Biological actions and therapeutic potential of the glucagon-like peptides. Gastroenterology122, 531–544.10.1053/gast.2002.31068Search in Google Scholar PubMed
Efrat, S. (1997). Making sense of glucose sensing. Nat. Genet.17, 249–250.10.1038/ng1197-249Search in Google Scholar PubMed
Ehses, J.A., Casilla, V.R., Doty, T., Pospisilik, J.A., Winter, K.D., Demuth, H.U., Pederson, R.A., and McIntosh, C.H. (2003). Glucose-dependent insulinotropic polypeptide promotes beta-(INS-1) cell survival via cyclic adenosine monophosphate-mediated caspase-3 inhibition and regulation of p38 mitogen-activated protein kinase. Endocrinology144, 4433–4445.10.1210/en.2002-0068Search in Google Scholar PubMed
Farilla, L., Hui, H., Bertolotto, C., Kang, E., Bulotta, A., Di Mario, U., and Perfetti, R. (2005). Glucagon-like peptide-1 promotes islet cell growth and inhibits apoptosis in Zucker diabetic rats. Endocrinology143, 4397–4408Search in Google Scholar
Flatt, P.R. and Bailey, C.J. (1981). Abnormal plasma glucose and insulin responses in heterozygous lean (ob/+) mice. Diabetologia20, 573–577.10.1007/BF00252768Search in Google Scholar
Gault, V.A., Flatt, P.R., Bailey, C.J., Harriott, P., Greer, B., Mooney, M.H., and O'Harte, F.P.M. (2002). Enhanced cyclic AMP generation and insulin-releasing potency of two novel N-terminal Tyr1-modified enzyme resistant forms of GIP, is associated with significant antihyperglycaemic activity in spontaneous obesity-diabetes. Biochem. J.367, 913–920.10.1042/bj20020319Search in Google Scholar
Gault, V.A., Flatt, P.R., and O'Harte, F.P. (2003). Glucose-dependent insulinotropic polypeptide analogues and their therapeutic potential for the treatment of obesity-diabetes. Biochem. Biophys. Res. Commun.308, 207–213.10.1016/S0006-291X(03)01361-5Search in Google Scholar
Green, B.D., Gault, V.A., O'Harte, F.P., and Flatt, P.R. (2004). Structurally modified analogues of glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) as future antidiabetic agents. Curr. Pharm. Des.10, 3651–3662.10.2174/1381612043382774Search in Google Scholar PubMed
Hansson, M., Tonning, A., Frandsen, U., Petri, A., Rajagopal, J., Englund, M.C., Heller, R.S., Hakansson, J., Fleckner, J., Skold, H.N., et al. (2004). Artifactual insulin release from differentiated embryonic stem cells. Diabetes53, 2603–2609.10.2337/diabetes.53.10.2603Search in Google Scholar PubMed
Hori, Y., Rulifson, I.C., Tsai, B.C., Heit, J.J., Cahoy, J.D., and Kim, S.K. (2002). Growth inhibitors promote differentiation of insulin-producing tissue from embryonic stem cells. Proc. Natl. Acad. Sci. USA99, 16105–16110.10.1073/pnas.252618999Search in Google Scholar PubMed PubMed Central
Lester, L.B., Kuo, H.C., Andrews, L., Nauert, B., and Wolf, D.P. (2004). Directed differentiation of rhesus monkey ES cells into pancreatic cell phenotypes. Reprod. Biol. Endocrinol.2, 42.10.1186/1477-7827-2-42Search in Google Scholar PubMed PubMed Central
Lipsett, M. and Finegood, D.T. (2002). β-Cell neogenesis during prolonged hyperglycemia in rats. Diabetes51, 1834–1841.10.2337/diabetes.51.6.1834Search in Google Scholar PubMed
Lumelsky, N., Blondel, O., Laeng, P., Velasco, I., Ravin, R., and McKay, R. (2001). Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets. Science292, 1389–1394.10.1126/science.1058866Search in Google Scholar PubMed
Mashima, H., Shibata, H., Mine, T., and Kojima, I. (1996). Formation of insulin-producing cells from pancreatic acinar AR42J cells by hepatocyte growth factor. Endocrinology137, 3969–3976.10.1210/endo.137.9.8756573Search in Google Scholar PubMed
Milne, H.M., Burns, C.J., Kitsou-Mylona, I., Luther, M.J., Minger, S.L., Persaud, S.J., and Jones, P.M. (2005). Generation of insulin-expressing cells from mouse embryonic stem cells. Biochem. Biophys. Res. Commun.328, 399–403.10.1016/j.bbrc.2004.12.183Search in Google Scholar PubMed
Moritoh, Y., Yamato, E., Yasui, Y., Miyazaki, S., and Miyazaki, J. (2003). Analysis of insulin-producing cells during in vitro differentiation from feeder-free embryonic stem cells. Diabetes52, 1163–1168.10.2337/diabetes.52.5.1163Search in Google Scholar PubMed
Rajagopal, J., Anderson, W.J., Kume, S., Martinez, O.I., and Melton, D.A. (2003). Insulin staining of ES cell progeny from insulin uptake. Science299, 363.10.1126/science.1077838Search in Google Scholar PubMed
Sander, M., Sussel, L., Conners, J., Scheel, D., Kalamaras, J., Dela, Cruz, F., Schwitzgebel, V., Hayes-Jordan, A., and German, M. (2000). Homeobox gene Nkx6.1 lies downstream of Nkx2.2 in the major pathway of β-cell formation in the pancreas. Development127, 5533–5540.Search in Google Scholar
Schmied, B.M., Ulrich, A., Matsuzaki, H., Ding, X., Ricordi, C., Weide, L., Moyer, M.P., Batra, S.K., Adrian, T.E., and Pour, P.M. (2001). Transdifferentiation of human islet cells in a long-term culture. Pancreas23, 157–171.10.1097/00006676-200108000-00006Search in Google Scholar PubMed
Segev, H., Fishman, B., Ziskind, A., Shulman, M., and Itskovitz-Eldor, J. (2004). Differentiation of human embryonic stem cells into insulin-producing clusters. Stem Cells22, 265–274.10.1634/stemcells.22-3-265Search in Google Scholar PubMed
Shapiro, A.M., Lakey, J.R., Ryan, E.A., Korbutt, G.S., Toth, E., Warnock, G.L., Kneteman, N.M., and Rajotte, R,V. (2000). Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. N. Engl. J. Med.343, 230–238.10.1056/NEJM200007273430401Search in Google Scholar PubMed
Shepherd, P.R. and Kahn, B.B. (1999). Glucose transporters and insulin action – implications for insulin resistance and diabetes mellitus. N. Engl. J. Med.341, 248–257.10.1056/NEJM199907223410406Search in Google Scholar PubMed
Sipione, S., Eshpeter, A., Lyon, J.G., Korbutt, G.S., and Bleackley, R.C. (2004). Insulin expressing cells from differentiated embryonic stem cells are not β cells. Diabetologia47, 499–508.10.1007/s00125-004-1349-zSearch in Google Scholar PubMed
Soria, B., Roche, E., Berna, G., Leon-Quinto, T., Reig, J.A., and Martin, F. (2000). Insulin-secreting cells derived from embryonic stem cells normalize glycemia in streptozotocin-induced diabetic mice. Diabetes49, 157–162.10.2337/diabetes.49.2.157Search in Google Scholar PubMed
Stoffers, D.A., Heller, R.S., Miller, C.P., and Habener, J.F. (1999). Developmental expression of the homeodomain protein IDX-1 in mice transgenic for an IDX-1 promoter/lacZ transcriptional reporter. Endocrinology140, 5374–5381.10.1210/endo.140.11.7122Search in Google Scholar PubMed
Zhou, J., Wang, X., Pineyro, M.A., and Egan, J.M. (1999). Glucagon-like peptide 1 and exendin-4 convert pancreatic AR42J cells into glucagon- and insulin-producing cells. Diabetes48, 2358–2366.10.2337/diabetes.48.12.2358Search in Google Scholar PubMed
©2006 by Walter de Gruyter Berlin New York
Articles in the same Issue
- 4th General Meeting of the International Proteolysis Society/International Conference on Protease Inhibitors
- Extracellular granzymes: current perspectives
- Impact of the N-terminal amino acid on targeted protein degradation
- Structural aspects of recently discovered viral deubiquitinating activities
- Cysteine cathepsins and caspases in silicosis
- The proprotein convertases and their implication in sterol and/or lipid metabolism
- PREPL: a putative novel oligopeptidase propelled into the limelight
- Human cathepsin L rescues the neurodegeneration and lethality in cathepsin B/L double-deficient mice
- Helicobacter pylori-induced downregulation of the secretory leukocyte protease inhibitor (SLPI) in gastric epithelial cell lines and its functional relevance for H. pylori-mediated diseases
- Increased local levels of granulocyte colony-stimulating factor are associated with the beneficial effect of pre-elafin (SKALP/trappin-2/WAP3) in experimental emphysema
- Interaction of a novel form of Pseudomonas aeruginosa alkaline protease (aeruginolysin) with interleukin-6 and interleukin-8
- Analysis of aldosterone-induced differential receptor-independent protein patterns using 2D-electrophoresis and mass spectrometry
- Modeling the 3D structure of wheat subtilisin/chymotrypsin inhibitor (WSCI). Probing the reactive site with two susceptible proteinases by time-course analysis and molecular dynamics simulations
- A stable analogue of glucose-dependent insulinotropic polypeptide, GIP(LysPAL16), enhances functional differentiation of mouse embryonic stem cells into cells expressing islet-specific genes and hormones
- Transcription factor FOXM1c is repressed by RB and activated by cyclin D1/Cdk4
- Despite its strong transactivation domain, transcription factor FOXM1c is kept almost inactive by two different inhibitory domains
- Inhibition of calcineurin by infusion of CsA causes hyperphosphorylation of tau and is accompanied by abnormal behavior in mice
- Isolation and properties of extracellular proteinases of Penicillium marneffei
- Isolation and comparative characterization of Ki-67 equivalent antibodies from the HuCAL® phage display library
Articles in the same Issue
- 4th General Meeting of the International Proteolysis Society/International Conference on Protease Inhibitors
- Extracellular granzymes: current perspectives
- Impact of the N-terminal amino acid on targeted protein degradation
- Structural aspects of recently discovered viral deubiquitinating activities
- Cysteine cathepsins and caspases in silicosis
- The proprotein convertases and their implication in sterol and/or lipid metabolism
- PREPL: a putative novel oligopeptidase propelled into the limelight
- Human cathepsin L rescues the neurodegeneration and lethality in cathepsin B/L double-deficient mice
- Helicobacter pylori-induced downregulation of the secretory leukocyte protease inhibitor (SLPI) in gastric epithelial cell lines and its functional relevance for H. pylori-mediated diseases
- Increased local levels of granulocyte colony-stimulating factor are associated with the beneficial effect of pre-elafin (SKALP/trappin-2/WAP3) in experimental emphysema
- Interaction of a novel form of Pseudomonas aeruginosa alkaline protease (aeruginolysin) with interleukin-6 and interleukin-8
- Analysis of aldosterone-induced differential receptor-independent protein patterns using 2D-electrophoresis and mass spectrometry
- Modeling the 3D structure of wheat subtilisin/chymotrypsin inhibitor (WSCI). Probing the reactive site with two susceptible proteinases by time-course analysis and molecular dynamics simulations
- A stable analogue of glucose-dependent insulinotropic polypeptide, GIP(LysPAL16), enhances functional differentiation of mouse embryonic stem cells into cells expressing islet-specific genes and hormones
- Transcription factor FOXM1c is repressed by RB and activated by cyclin D1/Cdk4
- Despite its strong transactivation domain, transcription factor FOXM1c is kept almost inactive by two different inhibitory domains
- Inhibition of calcineurin by infusion of CsA causes hyperphosphorylation of tau and is accompanied by abnormal behavior in mice
- Isolation and properties of extracellular proteinases of Penicillium marneffei
- Isolation and comparative characterization of Ki-67 equivalent antibodies from the HuCAL® phage display library