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.

  • Technical Report
  • Published:

A ligand-receptor fusion of growth hormone forms a dimer and is a potent long-acting agonist

Nature Medicine volume 13, pages 1108–1113 (2007)Cite this article

Abstract

Cytokine hormones have a short plasma half-life and require frequent administration. For example, growth hormone replacement involves daily injections. In common with other cytokines, the extracellular domain of the growth hormone receptor circulates as a binding protein, which naturally prolongs the biological half-life of growth hormone. Here we have studied the biological actions of a ligand-receptor fusion of growth hormone and the extracellular domain of its receptor. The genetically engineered ligand-receptor fusion protein was purified from mammalian cell culture. In rats, the ligand-receptor fusion had a 300-times reduced clearance as compared to native growth hormone, and a single injection promoted growth for 10 d, far exceeding the growth seen after administration of native growth hormone. The ligand-receptor fusion forms a reciprocal, head-to-tail dimer that provides a reservoir of inactive hormone similar to the natural reservoir of growth hormone and its binding protein. In conclusion, a ligand-receptor fusion of cytokine to its extracellular receptor generates a potent, long-acting agonist with exceptionally slow absorption and elimination. This approach could be easily applied to other cytokines.

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: Relationship between growth hormone, GHBP, the ligand-receptor (LR) fusion and the GHR on the basis of published structures27 (pdb3HHR).
Figure 2: Characterization and bioactivity of the ligand-receptor (LR) fusion.
Figure 3: Profiles of growth hormone (GH) and ligand-receptor (LR) fusion measured after subcutaneous and intravenous administration.
Figure 4: Body weight change in rats after subcutaneous administration of growth hormone (GH) and the ligand-receptor (LR) fusion.

Similar content being viewed by others

References

  1. Woodhouse, L.J., Mukherjee, A., Shalet, S.M. & Ezzat, S. The influence of growth hormone status on physical impairments, functional limitations, and health-related quality of life in adults. Endocr. Rev. 27, 287–317 (2006).

    Article  CAS  Google Scholar 

  2. Clark, R. et al. Long-acting growth hormones produced by conjugation with polyethylene glycol. J. Biol. Chem. 271, 21969–21977 (1996).

    Article  CAS  Google Scholar 

  3. Cook, D.M. et al. The pharmacokinetic and pharmacodynamic characteristics of a long-acting growth hormone (GH) preparation (nutropin depot) in GH-deficient adults. J. Clin. Endocrinol. Metab. 87, 4508–4514 (2002).

    Article  CAS  Google Scholar 

  4. Reiter, E.O. et al. A multicenter study of the efficacy and safety of sustained release GH in the treatment of naive pediatric patients with GH deficiency. J. Clin. Endocrinol. Metab. 86, 4700–4706 (2001).

    Article  CAS  Google Scholar 

  5. Jostel, A., Mukherjee, A., Alenfall, J., Smethurst, L. & Shalet, S.M. A new sustained-release preparation of human growth hormone and its pharmacokinetic, pharmacodynamic and safety profile. Clin. Endocrinol. 62, 623–627 (2005).

    Article  CAS  Google Scholar 

  6. Laursen, T. et al. Long-term effects of continuous subcutaneous infusion versus daily subcutaneous injections of growth hormone (GH) on the insulin-like growth factor system, insulin sensitivity, body composition, and bone and lipoprotein metabolism in GH-deficient adults. J. Clin. Endocrinol. Metab. 86, 1222–1228 (2001).

    CAS  PubMed  Google Scholar 

  7. Laursen, T., Jorgensen, J.O., Jakobsen, G., Hansen, B.L. & Christiansen, J.S. Continuous infusion versus daily injections of growth hormone (GH) for 4 weeks in GH-deficient patients. J. Clin. Endocrinol. Metab. 80, 2410–2418 (1995).

    PubMed  Google Scholar 

  8. Muller-Newen, G., Kohne, C. & Heinrich, P.C. Soluble receptors for cytokines and growth factors. Int. Arch. Allergy Immunol. 111, 99–106 (1996).

    Article  CAS  Google Scholar 

  9. Baumann, G., Amburn, K.D. & Buchanan, T.A. The effect of circulating growth hormone-binding protein on metabolic clearance, distribution, and degradation of human growth hormone. J. Clin. Endocrinol. Metab. 64, 657–660 (1987).

    Article  CAS  Google Scholar 

  10. Baumann, G. Growth hormone heterogeneity: genes, isohormones, variants, and binding proteins. Endocr. Rev. 12, 424–449 (1991).

    Article  CAS  Google Scholar 

  11. Baumann, G., Shaw, M.A. & Buchanan, T.A. In vivo kinetics of a covalent growth hormone-binding protein complex. Metabolism 38, 330–333 (1989).

    Article  CAS  Google Scholar 

  12. Clark, R.G. et al. Recombinant human growth hormone (GH)-binding protein enhances the growth-promoting activity of human GH in the rat. Endocrinology 137, 4308–4315 (1996).

    Article  CAS  Google Scholar 

  13. Baumann, G. Growth hormone binding protein—errant receptor or active player? Endocrinology 136, 377–378 (1995).

    Article  CAS  Google Scholar 

  14. Ayling, R.M. et al. A dominant-negative mutation of the growth hormone receptor causes familial short stature. Nat. Genet. 16, 13–14 (1997).

    Article  CAS  Google Scholar 

  15. Ross, R.J. et al. A short isoform of the human growth hormone receptor functions as a dominant negative inhibitor of the full-length receptor and generates large amounts of binding protein. Mol. Endocrinol. 11, 265–273 (1997).

    Article  CAS  Google Scholar 

  16. Ross, R.J.M. et al. Binding and functional studies with the growth hormone receptor antagonist, B2036-PEG (pegvisomant), reveal effects of pegylation and evidence that it binds to a receptor dimer. J. Clin. Endocrinol. Metab. 86, 1716–1723 (2001).

    CAS  PubMed  Google Scholar 

  17. Cunningham, B.C. et al. Dimerization of the extracellular domain of the human growth hormone receptor by a single hormone molecule. Science 254, 821–825 (1991).

    Article  CAS  Google Scholar 

  18. Huston, J.S., Tai, M.S., McCartney, J., Keck, P. & Oppermann, H. Antigen recognition and targeted delivery by the single-chain Fv. Cell Biophys. 22, 189–224 (1993).

    Article  CAS  Google Scholar 

  19. Herington, A.C., Smith, A.I., Wallace, C. & Stevenson, J.L. Partial purification from human serum of a specific binding protein for human growth hormone. Mol. Cell. Endocrinol. 53, 203–209 (1987).

    Article  CAS  Google Scholar 

  20. Frick, G.P., Tai, L.R., Baumbach, W.R. & Goodman, H.M. Tissue distribution, turnover, and glycosylation of the long and short growth hormone receptor isoforms in rat tissues. Endocrinology 139, 2824–2830 (1998).

    Article  CAS  Google Scholar 

  21. Mannor, D.A., Winer, L.M., Shaw, M.A. & Baumann, G. Plasma growth hormone (GH)-binding proteins: effect on GH binding to receptors and GH action. J. Clin. Endocrinol. Metab. 73, 30–34 (1991).

    Article  CAS  Google Scholar 

  22. Lim, L., Spencer, S.A., McKay, P. & Waters, M.J. Regulation of growth hormone (GH) bioactivity by a recombinant human GH-binding protein. Endocrinology 127, 1287–1291 (1990).

    Article  CAS  Google Scholar 

  23. Haffner, D., Schaefer, F., Girard, J., Ritz, E. & Mehls, O. Metabolic clearance of recombinant human growth hormone in health and chronic renal failure. J. Clin. Invest. 93, 1163–1171 (1994).

    Article  CAS  Google Scholar 

  24. Johnson, V. & Maack, T. Renal extraction, filtration, absorption, and catabolism of growth hormone. Am. J. Physiol. 233, F185–F196 (1977).

    CAS  PubMed  Google Scholar 

  25. Veldhuis, J.D. et al. Impact of experimental blockade of peripheral growth hormone (GH) receptors on the kinetics of endogenous and exogenous GH removal in healthy women and men. J. Clin. Endocrinol. Metab. 87, 5737–5745 (2002).

    Article  CAS  Google Scholar 

  26. Osborn, B.L. et al. Albutropin: a growth hormone-albumin fusion with improved pharmacokinetics and pharmacodynamics in rats and monkeys. Eur. J. Pharmacol. 456, 149–158 (2002).

    Article  CAS  Google Scholar 

  27. de Vos, A.M., Ultsch, M. & Kossiakoff, A.A. Human growth hormone and extracellular domain of its receptor: crystal structure of the complex. Science 255, 306–312 (1992).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to ARC BioServ and particularly to S. Smith and I. Phillips for their support in protein production. This work was supported through an Asterion Ltd. contract with the University of Sheffield.

Author information

Author notes

  1. Ian R Wilkinson, Eric Ferrandis and Peter J Artymiuk: These authors contributed equally to this work.

Authors and Affiliations

  1. School of Medicine and Biomedical Sciences, Royal Hallamshire Hospital, University of Sheffield, Sheffield, S10 2JF, UK

    Ian R Wilkinson, Sarbendra L Pradhananga, Sue Justice & Richard J Ross

  2. Institut Henri Beaufour, Ipsen, 5 Avenue du Canada, Les Ulis Cedex, 91966, France

    Eric Ferrandis, Marc Teillot, Chantal Soulard & Caroline Touvay

  3. Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, S10 2JF, UK

    Peter J Artymiuk

  4. Department of Medicine, Campus Mitte Charite-Universitatsmedizim, Berlin, 10117, Germany

    Zida Wu & Christian J Strasburger

  5. Pituitary Research Unit, Garvan Institute of Medical Research, Sydney, 2010, New South Wales, Australia

    Kin C Leung

  6. Unit of Infection and Immunity, Royal Hallamshire Hospital, University of Sheffield, Sheffield, S10 2JF, UK

    Jon R Sayers

Authors
  1. Ian R Wilkinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Eric Ferrandis
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Peter J Artymiuk
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Marc Teillot
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chantal Soulard
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Caroline Touvay
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Sarbendra L Pradhananga
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Sue Justice
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Zida Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Kin C Leung
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Christian J Strasburger
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Jon R Sayers
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Richard J Ross
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

I.R.W., S.L.P. and S.J. cloned, purified, tested and analyzed the fusion proteins. Z.W., K.C.L. and C.J.S. contributed to the characterization of the fusion proteins. E.F., M.T., C.S. and C.T. performed and analyzed the in vivo studies. E.F., P.J.A., J.R.S. and R.J.R. were responsible for the concepts, data analysis and writing of the manuscript. R.J.R. supervised the project.

Corresponding author

Correspondence to Richard J Ross.

Ethics declarations

Competing interests

S.J., Z.W., K.C.L. and C.J.S. have no competing interests to declare. I.W., P.J.A., S.L.P., J.R.S. and R.J.R. have an equity interest in Asterion Ltd., and P.J.A., J.R.S. and R.J.R. are directors of Asterion Ltd. E.F., M.T., C.S. and C.T. are employed by Ipsen. This work was supported through an Asterion Ltd. contract with the University of Sheffield.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wilkinson, I., Ferrandis, E., Artymiuk, P. et al. A ligand-receptor fusion of growth hormone forms a dimer and is a potent long-acting agonist. Nat Med 13, 1108–1113 (2007). https://doi.org/10.1038/nm1610

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm1610

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing