FGF-21 as a novel metabolic regulator

Alexei Kharitonenkov

Tatiyana Shiyanova

Anja Koester

Amy Ford

Julie Moyers+9 authors

^{Lilly Research Laboratories, Division of Eli Lilly and Co., Indianapolis, Indiana, USA.}^{Lilly Research Laboratories, Hamburg, Germany.}

Address correspondence to: Alexei Kharitonenkov, Lilly Research Laboratories, Division of Eli Lilly and Company, Indianapolis, Indiana 46285, USA. Phone: (317) 276-0091; Fax: (317) 277-2934; E-mail: moc.yllil@hcrahk.a.

Received 2004 Oct 12; Accepted 2005 Mar 23.

Abstract

Diabetes mellitus is a major health concern, affecting more than 5% of the population. Here we describe a potential novel therapeutic agent for this disease, FGF-21, which was discovered to be a potent regulator of glucose uptake in mouse 3T3-L1 and primary human adipocytes. FGF-21–transgenic mice were viable and resistant to diet-induced obesity. Therapeutic administration of FGF-21 reduced plasma glucose and triglycerides to near normal levels in both ob/ob and db/db mice. These effects persisted for at least 24 hours following the cessation of FGF-21 administration. Importantly, FGF-21 did not induce mitogenicity, hypoglycemia, or weight gain at any dose tested in diabetic or healthy animals or when overexpressed in transgenic mice. Thus, we conclude that FGF-21, which we have identified as a novel metabolic factor, exhibits the therapeutic characteristics necessary for an effective treatment of diabetes.

Abstract

Acknowledgments

We wish to thank P. Atkinson, G. Kelly, T. Black, B. Pies, and B. Strifler for supporting FGF-21 protein production; D. Bruce Baldwin for the generation of FGFR-FC-fusion constructs; S. Bright and J. Dunbar for FGF-21/FGFR binding experiments; S. Sissons and W. Roell for assistance with glucose uptake and proliferation assays; N. Fox and K. Brune for generation of FGF-21–transgenic animals; K. Coble for help with pharmacokinetic studies; M. Brenner and A. Efanov for assistance with glucagon secretion experiments; D. Ballard and K. Mintze for supporting histochemistry work; C. Shrake for in vivo assistance; J. Manetta and L. Slieker for generation of polyclonal anti–FGF-21, anti-GLUT1, and anti-GLUT4 antibodies; A. Klip (Hospital for Sick Children, Toronto, Ontario, Canada) for L6-GLUT-4myc cells; J. Caro, S. Jacobs, and G. Etgen for critically reading the manuscript; and T. Bumol, B. Grinnell, A. Glasebrook, R. Smith, and S. Taylor for helpful discussions.

Acknowledgments

Footnotes

Nonstandard abbreviations used: BAT, brown adipose tissue; BMP-9, bone morphogenic protein–9; EC₅₀, 50% effective concentration; FGFR, FGF receptor; FRS-2, FGFR substrate–2; GLP-1, glucagon-like peptide–1; GLUT, glucose transporter; HFHC, high-fat/high-carbohydrate diet; HMEC, primary human mammary epithelial cell; HUVEC, human umbilical vein endothelial cell; OGTT, oral glucose tolerance test; PCNA, proliferative cell nuclear antigen; ZDF, Zucker diabetic fatty.

Conflict of interest: The authors have declared that no conflict of interest exists.

Footnotes

References

1. McKeehan WL, Wang F, Kan MThe heparan sulfate-fibroblast growth factor family: diversity of structure and function. Prog. Nucleic Acid Res. Mol. Biol.1998;59:135–176.[PubMed]
2. Galzie Z, Kinsella AR, Smith JAFibroblast growth factors and their receptors. Biochem. Cell Biol.1997;75:669–685.[PubMed]
3. Powers CJ, McLeskey SW, Wellstein AFibroblast growth factors, their receptors and signaling. Endocr. Relat. Cancer.2000;7:165–197.[PubMed]
4. Hogan BLMorphogenesis. Cell.1999;96:225–233.[PubMed]
5. Ornitz DM, Itoh NFibroblast growth factors. Genome Biol.2001;2:reviews3005. doi:10.1186/gb–2001-2-3-reviews3005.
6. Sakaue H, et al Requirement of fibroblast growth factor 10 in development of white adipose tissue. Genes Dev.2002;16:908–912.
7. Bhushan A, et al FGF10 is essential for maintaining the proliferative capacity of epithelial progenitor cells during early pancreatic organogenesis. Development.2001;128:5109–5117.[PubMed]
8. Ohuchi H, et al FGF10 acts as a major ligand for FGF receptor 2 IIIb in mouse multi-organ development. Biochem. Biophys. Res. Commun.2000;277:643–649.[PubMed]
9. Konishi M, Micami T, Yamasaki M, Miyake A, Itoh NFibroblast growth factor-16 is a growth factor for embryonic brown adipocytes. J. Biol. Chem.1999;255:12119–12122.[PubMed]
10. Nishimura T, Utsonomiya Y, Hoshikawa M, Ohuchi H, Itoh NStructure and expression of a novel human FGF, FGF-19, expressed in the fetal brain. . Biochim. Biophys. Acta.1999; 1444:148–151.[PubMed]
11. Xie MH, et al FGF-19, a novel fibroblast growth factor with unique specificity for FGFR4. Cytokine.1999;11:729–735.[PubMed]
12. Tomlinson E, et al Transgenic mice expressing human fibroblast growth factor-19 display increased metabolic rate and decreased adiposity. Endocrinology.2002;143:1741–1747.[PubMed]
13. Fu L, et al Fibroblast growth factor 19 increases metabolic rate and reverses dietary and leptin deficient diabetes. Endocrinology.2004;145:2594–2603.[PubMed]
14. Kim I, Moon S, Yu K, Kim U, Koh GYA novel fibroblast growth factor receptor-5 preferentially expressed in the pancreas. . Biochim. Biophys. Acta.2001; 1518:152–156.[PubMed]
15. Sleeman M, et al Identification of a novel fibroblast growth factor receptor, FGFR5. Gene.2001;271:171–182.[PubMed]
16. Hart AW, Baeza N, Apelqvist Å, Edlund HAttenuation of FGF-signalling in mouse β-cells leads to diabetes. Nature.2000;408:864–868.[PubMed]
17. Revest JM, et al Fibroblast growth factor receptor 2 IIIb acts upstream of Shh and FGF4 and is required for limb bud maintenance but not for the induction of FGF8, FGF10, Msx1, or BMP4. Dev. Biol.2001;231:47–62.[PubMed]
18. Celli G, LaRochelle WJ, Mackem S, Sharp R, Merlino GSoluble dominant-negative receptor uncovers essential roles for fibroblast growth factors in multi-organ induction and patterning. EMBO J.1998;17:1642–1655.
19. Elghazi L, Cras-Meneur C, Czernichow P, Scharfmann RRole for FGFR2IIIb-mediated signals in controlling pancreatic endocrine progenitor cell proliferation. Proc. Natl. Acad. Sci. U. S. A.2002;99:3884–3889.
20. Yu C, et al Elevated cholesterol metabolism and bile acid synthesis in mice lacking membrane tyrosine kinase receptor FGFR4. J. Biol. Chem.2000;275:15482–15489.[PubMed]
21. Nishimura T, Nakatake Y, Konishi M, Itoh NIdentification of a novel FGF, FGF-21, preferentially expressed in the liver. . Biochim. Biophys. Acta.2000; 1492:203–206.[PubMed]
22. Ueyama A, Yaworsky KL, Wang Q, Ebina Y, Klip AGLUT-4myc ectopic expression in L6 myoblasts generates a GLUT-4-specific pool conferring insulin sensitivity. Am. J. Physiol.1999;277:572–578.[PubMed]
23. Lin W-W, Hsu Y-WCycloheximide-induced cPLA₂ activation is via the MPK-1 down-regulation and ERK activation. Cell. Signal.2000;12:457–461.[PubMed]
24. Kouhara H, et al A lipid-anchored GRB2-binding protein that links FGF-receptor activation to the Ras/MAPK signalling pathway. Cell.1997;89:693–702.[PubMed]
25. Schlessinger JCell signalling by receptor tyrosine kinases. Cell.2000;103:211–225.[PubMed]
26. Nicholes K, et al. A mouse model of hepatocellular carcinoma. Ectopic expression of fibroblast growth factor 19 in skeletal muscle of transgenic mice. Am. J. Pathol.2002;160:2295–2307.
27. Moller DENew drug targets for type 2 diabetes and the metabolic syndrome. Nature.2001;414:821–827.[PubMed]
28. Chen C, et al An integrated functional genomics screening program reveals a role for BMP-9 in glucose homeostasis. Nat. Biotechnol.2003;21:294–301.[PubMed]
29. Shepherd PR, Khan BBGlucose transporters and insulin action. N. Eng. J. Med.1999;341:248–257.[PubMed]
30. Coulier F, et al Of worms and men: an evolutionary perspective on the fibroblast growth factor (FGF) and FGF receptor families. J. Mol. Evol.1997;44:43–56.[PubMed]
31. Frederich RC, et al Leptin levels reflect body lipid content in mice: evidence for diet-induced resistance to leptin action. Nat. Med.1995;1:1311–1314.[PubMed]
32. Linden MD, Torres FX, Kubus MS, Zarbo RJClinical application of morphologic and immunocytochemical assessments of cell proliferation. Am. J. Clin. Pathol.1992;97:S4–S13.[PubMed]
33. Reaven GM, Chen YD, Golay A, Swislocki AL, Jaspan JBDocumentation of hyperglucagononemia throughout the day in nonobese and obese patients with noninsulin-dependent diabetes mellitus. J. Clin. Endocrinol. Metab.1987;64:106–110.[PubMed]
34. Shah PA, et al Lack of suppression of glucagon contributes to postprandial hyperglycemia in subjects with type 2 diabetes mellitus. J. Clin. Endocrinol. Metab.2000;85:4053–4059.[PubMed]
35. Sloop KW, et al Hepatic and glucagon-like peptide-1-mediated reversal of diabetes by glucagon receptor antisense oligonucleotide inhibitors. J. Clin. Invest.2004;113:1571–1581. doi:10.1172/JCI200420911.
36. Zimmet P, Alberti KG, Shaw JGlobal and societal implications of the diabetes epidemic. Nature.2001;414:782–787.[PubMed]
37. Taylor SIDeconstructing type 2 diabetes. Cell.1999;97:9–12.[PubMed]
38. Saltiel ARNew perspectives into the molecular pathogenesis and treatment of type 2 diabetes. Cell.2001;104:517–529.[PubMed]
39. Cheatham B, et al Phosphatidylinositol 3-kinase activation is required for insulin stimulation of pp70 S6 kinase, DNA synthesis, and glucose transporter translocation. Mol. Cell. Biol.1994;14:4902–4911.
40. Kharitonenkov A, et al A family of proteins that inhibit signalling through tyrosine kinase receptors. Nature.1997;386:181–186.[PubMed]
41. Slieker LJ, et al Glucose transporter levels in tissues of spontaneously diabetic Zucker fa/fa rat (ZDF/drt) and viable yellow mouse (Avy/a) Diabetes.1992;41:187–193.[PubMed]
42. Allan CM, Walker D, Taylor JMEvolutionary duplication of a hepatic control region in the human apolipoprotein E gene locus. J. Biol. Chem.1995;270:26278–26281.[PubMed]
43. Hogan, B., Beddington, R., Constantini, F., and Laci, E1994. Manipulating the mouse embryo. Cold Spring Harbor Press. Cold Spring Harbor, New York, USA. 487 pp.