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.

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.


Yang Z, Han S, Keller M, Kaiser A, Bender BJ, Bosse M, Burkert K, Kögler LM, Wifling D, Bernhardt G, Plank N, Littmann T, Schmidt P, Yi C, Li B, Ye S, Zhang R, Xu B, Larhammar D, Stevens RC, Huster D, Meiler J, Zhao Q, Beck-Sickinger AG, Buschauer A, Wu B. Structural basis of ligand binding modes at the neuropeptide Y Y1 receptor. Nature. 2018 Apr;556(7702):520-524.


Burkert K, Zellmann T, Meier R, Kaiser A, Stichel J, Meiler J, Mittapalli GK, Roberts E, Beck-Sickinger AG. A Deep Hydrophobic Binding Cavity is the Main Interaction for Different Y(2) R Antagonists. ChemMedChem. 2016; Epub ahead of print.


Wanka L, Babilon S, Burkert K, Mörl K, Gurevich VV, Beck-Sickinger AG. C-terminal motif of human neuropeptide Y4 receptor determines internalization and arrestin recruitment. Cell Signal. 2017;29:233-9.


Thieme V, Jolly N, Madsen AN, Bellmann-Sickert K, Schwartz TW, Holst B, Cox HM,Beck-Sickinger AG. High molecular weight PEGylation of human pancreatic polypeptide at position 22 improves stability and reduces food intake in mice. Br J Pharmacol. 2016; Epub ahead of print.


Park M, Sivertsen BB, Els-Heindl S, Huber T, Holst B, Beck-Sickinger AG, Schwartz TW, Sakmar TP. Bioorthogonal labeling of ghrelin receptor to facilitate studies of ligand-dependent conformational dynamics. Chem Biol. 2015;22:1431-6.


Kaiser A, Müller P, Zellmann T, Scheidt HA, Thomas L, Bosse M, Meier R, Meiler J, Huster D, Beck-Sickinger AG, Schmit P. Unwinding of the C-terminal residues of neuropeptide Y is critical for Y2 receptor binding and activation. Angew Chem Int Ed Engl. 2015;54:7446-9.


Kilian TM, Klöting N, Bergmann R, Els-Heindl S, Babilon S, Clement-Ziza M, Zhang Y, Beck-Sickinger AG, Chollet C. Rational design of dual peptides targeting ghrelin and Y2 receptors to regulate food intake and body weight. J Med Chem. 2015;58:4180-9.


Kostelnik KB, Els-Heindl S, Klöting N, Baumann S, von Bergen M, Beck-Sickinger AG. High metabolic in vivo stability and bioavailability of a palmitoylated ghrelin receptor ligand assessed by mass spectrometry. Bioorg Med Chem. 2015;23:3925-32.


Rojas JM, Bruinstroop E, Printz RL, Alijagic-Boers A, Foppen E, Turney MK, George L, Beck-Sickinger AG, Kalsbeek A, Niswender KD. entral nervous system neuropeptide Y regulates mediators of hepatic phospholipid remodeling and very low-density lipoprotein triglyceride secretion via sympathetic innervation. Mol Metab. 2015;4:210-21.


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.


Mäde V, Babilon S, Jolly N, Wanka L, Bellmann-Sickert K, Diaz Gimenez LE, Mörl K, Cox HM, Gurevich VV, Beck-Sickinger AG. Peptide modifications differentially alter G protein-coupled receptor internalization and signaling bias. Angew Chem Int Ed. 2014;53:10067-71.


Mäde V, Els-Heindl S, Beck-Sickinger AG. Automated solid-phase peptide synthesis to obtain therapeutic peptides. Beilstein J Org Chem. 2014;110:1197-212.


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.


Walther C, Nagel S, Gimenez LE, Mörl K, Gurevich VV, Beck-Sickinger AG. Ligand-induced internalization and recycling of the human neuropeptide Y2 receptor is regulated by its carboxyl-terminal tail. J Biol Chem. 2010;285:41578-90.


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.


Lindner D, van Dieck J, Merten N, Mörl K, Günther R, Hofmann HJ, Beck-Sickinger AG. GPC receptors and not ligands decide the binding mode in neuropeptide Y multireceptor/multiligand system. Biochemistry. 2008;47:5905-14.


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.

 

PROJECT TEAM

Miron Gershkovich, Phd student

Sarina Rudolf, Phd student

Isabelle Ziffert, Phd student

Mareike Schenk, Phd student

Corinna Schüß, Phd student

Victoria Behr, Phd student