Abstract
Glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) are regarded as ‘incretins’ working closely to regulate glucose homeostasis. Unimolecular dual and triple agonists of GLP-1R and GIPR have shown remarkable clinical benefits in treating type 2 diabetes. However, their pharmacological characterization is usually carried out in a single receptor-expressing system. In the present study we constructed a co-expression system of both GLP-1R and GIPR to study the signaling profiles elicited by mono, dual and triple agonists. We show that when the two receptors were co-expressed in HEK 293T cells with comparable receptor ratio to pancreatic cancer cells, GIP predominately induced cAMP accumulation while GLP-1 was biased towards β-arrestin 2 recruitment. The presence of GIPR negatively impacted GLP-1R-mediated cAMP and β-arrestin 2 responses. While sharing some common modulating features, dual agonists (peptide 19 and LY3298176) and a triple agonist displayed differentiated signaling profiles as well as negative impact on the heteromerization that may help interpret their superior clinical efficacies.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Baggio LL, Drucker DJ. Biology of incretins: GLP-1 and GIP. Gastroenterology. 2007;132:2131–57.
Holst JJ. The incretin system in healthy humans: the role of GIP and GLP-1. Metabolism. 2019;96:46–55.
Seino Y, Fukushima M, Yabe D. GIP and GLP-1, the two incretin hormones: Similarities and differences. J Diabetes Investig. 2010;1:8–23.
Yamada Y, Miyawaki K, Tsukiyama K, Harada N, Yamada C, Seino Y. Pancreatic and extrapancreatic effects of gastric inhibitory polypeptide. Diabetes. 2006;125:170201.
Buhren BA, Gasis M, Thorens B, Müller HW, Bosse F. Glucose-dependent insulinotropic polypeptide (GIP) and its receptor (GIPR): Cellular localization, lesion-affected expression, and impaired regenerative axonal growth. J Neurosci Res. 2010;87:1858–70.
Nagashima M, Watanabe T, Terasaki M, Tomoyasu M, Nohtomi K, Kim-Kaneyama J, et al. Native incretins prevent the development of atherosclerotic lesions in apolipoprotein e knockout mice. Diabetologia. 2011;54:2649–59.
Fujita Y, Wideman RD, Asadi A, Yang GK, Baker R, Webber T, et al. Glucose-dependent insulinotropic polypeptide is expressed in pancreatic islet α-cells and promotes insulin secretion. Gastroenterology. 2010;138:1966–75.
Scrocchi LA, Brown TJ, Maclusky N, Brubaker PL, Auerbachet AB. Glucose intolerance but normal satiety in mice with a null mutation in the glucagon-like peptide 1 receptor gene. Nat Med. 1996;2:1254–8.
Sánchez-Garrido MA, Brandt SJ, Clemmensen C, Müller TD, Tschöp MH. Glp-1/glucagon receptor co-agonism for treatment of obesity. Diabetologia. 2017;60:1851–61.
Jones IR, Owens DR, Luzio S, Williams S, Hayes TM. The glucose-dependent insulinotropic polypeptide response to oral glucose and mixed meals is increased in patients with type 2 (non-insulin-dependent) diabetes mellitus. Diabetologia. 1989;32:668–77.
Finan B, Tao M, Ottaway N, Müller TD, Tschöp MH. Unimolecular dual incretins maximize metabolic benefits in rodents, monkeys, and humans. Sci Transl Med. 2013;209:151.
Jall S, Sachs S, Clemmensen C, Finan B, Neff F, Dimarchi RD, et al. Monomeric GLP-1/GIP/glucagon triagonism corrects obesity, hepatosteatosis, and dyslipidemia in female mice. Mol Metab. 2017;6:440–6.
Frias JP, Bastyr EJ, Vignati L, Tschöp MH, Schmitt C, Owen K, et al. The sustained effects of a dual GIP/GLP-1 receptor agonist, NNC0090-2746, in patients with type 2 diabetes. Cell Metab. 2017;26:343–52.
Bastin M, Andreelli F. Dual GIP–GLP1-receptor agonists in the treatment of type 2 diabetes: a short review on emerging data and therapeutic potential. Diabetes Metab Syndr Obes Targets Ther. 2019;12:1973–85.
Frias JP, Nauck MA, Van J, Kutner ME, Cui X, Benson C. Efficacy and safety of LY3298176, a novel dual GIP and GLP-1 receptor agonist, in patients with type 2 diabetes: a randomized, placebo-controlled and active comparator-controlled phase 2 trial. Lancet. 2018;392:2180–93.
Yabe D, Seino Y. Two incretin hormones GLP-1 and GIP: Comparison of their actions in insulin secretion and β cell preservation. Prog Biophys Mol Biol. 2011;107:248–56.
de Graaf C, Donnelly D, Wootten D, Lau J, Sexton PM, Miller LJ, et al. Glucagon-like peptide-1 and its class B G protein-coupled receptors: a long march to therapeutic successes. Pharmacol Rev. 2016;68:954–1013.
Skow MA, Bergmann NC, Knop FK. Diabetes and obesity treatment based on dual incretin receptor activation: ‘twincretins’. Diabetes Obes Metab. 2016;18:847–54.
Harikumar KG, Morfis MM, Sexton PM, Miller LJ. Pattern of intra-family hetero-oligomerization involving the G-protein-coupled secretin receptor. J Mol Neurosci. 2008;36:279–85.
Song X, Yu Y, Shen C, Wang Y, Wang N. Dimerization/oligomerization of the extracellular domain of the GLP-1 receptor and the negative cooperativity in its ligand binding revealed by the improved NanoBiT. FASEB J. 2020;34:4348–68.
Schelshorn D, Joly F, Mutel S, Hampe C, Breton B, Mutel V, et al. Lateral allosterism in the glucagon receptor family: Glucagon-like peptide 1 induces G-protein-coupled receptor heteromer formation. Mol Pharmacol. 2012;81:309–18.
Roed SN, Nhr AC, Wismann P, Iversen H, Brner-Osborne H, Knudsen SM, et al. Functional consequences of glucagon-like peptide-1 receptor cross-talk and trafficking. J Biol Chem. 2015;290:1233–43.
Zhao P, Liang YL, Belousoff MJ, Deganutti G, Wootten D. Activation of the GLP-1 receptor by a non-peptidic agonist. Nature. 2020;577:1–5.
Dal Maso E, et al. Extracellular loops 2 and 3 of the calcitonin receptor selectively modify agonist binding and efficacy. Biochem Pharmacol. 2018;150:214–44.
Yuliantie E, Darbalaei S, Dai A, Zhao P, Yang D, Sexton PM, et al. Pharmacological characterization of mono-, dual- and tri-peptidic agonists at GIP and GLP-1 receptors. Biochem Pharmacol. 2020;177:114001.
Wootten D, Miller LJ. Structural basis for allosteric modulation of Class B G protein-coupled receptors. Annu Rev Pharmacol Toxicol. 2020;60:89–107.
Al-Sabah S. Molecular pharmacology of the incretin receptors. Med Princ Pract. 2016;25:15–21.
Pederson RA, Satkunarajah M, Mcintosh CH, Scrocchi LA, Flamez D, Schuit F, et al. Enhanced glucose-dependent insulinotropic polypeptide secretion and insulinotropic action in glucagon-like peptide 1 receptor −/− mice. Diabetes. 1998;47:1046–52.
Miyawaki K, Yamada Y, Yano H, Niwa H, Ban N, Yu I, et al. Glucose intolerance caused by a defect in the entero-insular axis: a study in gastric inhibitory polypeptide receptor knockout mice. Proc Natl Acad Sci USA. 1999;96:14843–7.
Nauck MA. Normalization of fasting hyperglycaemia by exogenous glucagon-like peptide 1 (7-36 amide) in type 2 (non-insulin-dependent) diabetic patients. Diabetologia. 1993;36:741–4.
Tschöp M, Finan B, Clemmensen C, Gelfanov V, Dimarchi R, et al. Unimolecular polypharmacy for treatment of diabetes and obesity. Cell Metab. 2016;24:51–62.
Kang DS, Tian X, Benovic JL. Role of β-arrestins and arrestin domain-containing proteins in G protein-coupled receptor trafficking. Curr Opin Cell Biol. 2014;27:63–71.
Lefkowitz RJ, Pierce KL, Luttrell LM. Dancing with different partners: protein kinase A phosphorylation of seven membrane-spanning receptors regulates their G protein-coupling specificity. Mol Pharmacol. 2002;62:971–4.
Ferré S. The GPCR heterotetramer: challenging classical pharmacology. Trends Pharmacol Sci. 2015;36:145–52.
Kleinau G, Müller A, Biebermann H. Oligomerization of GPCRs involved in endocrine regulation. J Mol Endocrinol. 2016;57:59–80.
Acknowledgements
We thank Lijun Shao for technical assistance. This work was partially supported by National Natural Science Foundation of China 81872915 (MWW), 82073904 (MWW), 81773792 (DHY) and 81973373 (DHY); National Science and Technology Major Project of China–Key New Drug Creation and Manufacturing Program 2018ZX09735–001 (MWW) and 2018ZX09711002–002–005 (DHY); the National Key Basic Research Program of China 2018YFA0507000 (MWW); Novo Nordisk-CAS Research Fund grant NNCAS-2017–1-CC (DHY); and SA-SIBS Scholarship Program (DHY).
Author information
Authors and Affiliations
Contributions
YZW designed expression constructs, conducted cell-based assays, analyzed the data, and drafted the manuscript. DHY supervised the experiments, analyzed the data, and edited the manuscript. MWW conceived the idea, designed the studies, and edited the manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Supplementary information
Rights and permissions
About this article
Cite this article
Wang, Yz., Yang, Dh. & Wang, Mw. Signaling profiles in HEK 293T cells co-expressing GLP-1 and GIP receptors. Acta Pharmacol Sin 43, 1453–1460 (2022). https://doi.org/10.1038/s41401-021-00758-6
Received:
Accepted:
Published:
Version of record:
Issue date:
DOI: https://doi.org/10.1038/s41401-021-00758-6
Keywords
This article is cited by
-
Endogenous cell membrane interactome mapping for the GLP-1 receptor in different cell types
Nature Chemical Biology (2025)
