A3 - Role of Y-Receptors in the Regulation of Food Intake
At present, the modulation of satiety and hunger hormone activity is a promising approach in the development of new anti-obesity pharmaceuticals. Members of the Neuropeptide Y hormone family (Neuropeptide Y, Pancreatic Polypeptide and Peptide YY) are key player in the central and peripheral regulation of food intake, mediated by Gi protein coupled receptors Y1, Y2, Y4, und Y5. Since agonists are needed for all satiety hormones, receptor internalization has to be taken into account, as it may contribute to cell surface receptor desensitization and activation of alternative signaling pathways. Consequently, to understand the mechanisms and therapeutic potential of agonistic ligands, receptor internalization has to be studied in detail.
Aim of the project is the in vitro characterization of cellular effects influenced by Y-receptor subtype specific internalization, we could previously demonstrate, as well as the analysis of its contribution to physiological functions in vivo. We hypothesize that tissue and subtype specific Y-receptor signaling and internalization contribute to the central and peripheral regulation of food intake, thereby influencing the therapeutic potential of agonistic peptides. We will compare agonistic peptide ligands, which provoke altered receptor internalization and recycling as compared to the endogenous ligand NPY, with respect to cell specific internalization and signaling. Following the characterization of internalization patterns in hypothalamic cell lines, we will compare ligand induced expression profiles. Furthermore, we will evaluate the physiological effects induced in vivo after intracerebroventricular injection of these peptides, to understand the significance of receptor internalization in vivo zu verstehen.
Abbildung 1: Characterization of the ligands NPY, [F7,P34] NPY and [Gly34] NPY . Left: All peptides show comparable induction of G-protein signalling. Right: HEK293 cells stably expressing Y1-YFP fusionproteins (Yellow). Incubation with the respective TAM labelled peptides (red) for 60 min induces different receptor internalization. Nuclei are stained in blue.
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Kaiser A, Wanka L, Ziffert I, Beck-Sickinger AG. Biased agonists at the human Y1 receptor lead to prolonged membrane residency and extended receptor G protein interaction. Cell Mol Life Sci. 2020 Jan 9. doi: 10.1007/s00018-019-03432-7.
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Kögler LM, Stichel J, Kaiser A, Beck-Sickinger AG. Cell-Free Expression and Photo-Crosslinking of the Human Neuropeptide Y2 Receptor. Front Pharmacol. 2019 Mar 1;10:176.
Wanka L, Babilon S, Kaiser A, Mörl K, Beck-Sickinger AG.Different mode of arrestin-3 binding at the human Y1 and Y2 receptor.Cell Signal. 2018 Jun 23. pii: S0898-6568(18)30130-X.
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Pedragosa-Badia X, Sliwoski GR, Dong Nguyen E, Lindner D, Stichel J, Kaufmann KW, Meiler J, Beck-Sickinger AG. Pancreatic polypeptide is recognized by two hydrophobic domains onf the human Y4 receptor binding pockelt. J Biol Chem. 2014;289:5846-59.
Diaz Gimenez LE, Babilon S, Wanka L, Beck-Sickinger AG, Gurevich VV. Mutations in arrestin-3 differentially affect binding to neuropeptide Y receptor subtypes. Cell Signal. 2014;26:1523-31.
Mäde V, Bellmann-Sickert K, Kaiser A, Meiler J, Beck-Sickinger AG. Position and length of fatty acids strongly affect receptor selectivity pattern of human pancreatic polypeptide analogues. ChemMedChem. 2014;9:2463-74.
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Klöting N, Kovacs P, Kern M, Heiker JT, Fasshauer M, Schön MR, Stumvoll M, Beck-Sickinger AG, Blüher M. Central vaspin administration acutely reduces food intake and has sustained blood glucose-lowering effects. Diabetologia. 2011;54:1819-23.
Böhme I, Stichel J, Walther C, Mörl K, Beck-Sickinger AG. Agonist induced receptor internalization of neuropeptide Y receptor subtypes depends on third intracellular loop and C-terminus. Cell Signal. 2008;20:1740-9.
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Khan IU, Zwanziger D, Böhme I, Javed M, Naseer H, Hyder SW, Beck-Sickinger AG. Breast-cancer diagnosis by neuropeptide Y analogues: from synthesis to clinical application. Angew Chem Int Ed Engl. 2010;49:1155-8.
Keller M, Pop N, Hutzler C, Beck-Sickinger AG, Bernhardt G, Buschauer A. Guanidine-acylguanidinebioisosteric approach in the design of radioligands: synthesis of a tritium-labeled N(G)-propionylargininamide ([3H]-UR-MK114) as a highly potent and selective neuropeptide Y Y1 receptor antagonist. J Med Chem. 2008;51:8168-72.
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Merten N, Lindner D, Rabe N, Römpler H, Mörl K, Schöneberg T, Beck-Sickinger AG. Receptor subtype-specific docking of Asp6.59 with C-terminal arginine residues in Y receptor ligands. J Biol Chem. 2007;282:7543-51.
Dinger MC, Bader JE, Kobor AD, Kretzschmar AK, Beck-Sickinger AG. Homodimerization of neuropeptide y receptors investigated by fluorescence resonance energy transfer in living cells. J Biol Chem. 2003;278:10562-71.