Effects of Monoclonal Antagonist Antibodies on Calcitonin Gene-related Peptide Receptor Function and Trafficking

Overview

Journal J Headache Pain

Publisher Biomed Central

Date 2019 May 2

PMID 31039731

Citations 14

Authors

Raffi Manoukian

Hong Sun

Silke Miller

Di Shi

Brian Chan

Cen Xu

Affiliations

Soon will be listed here.

Abstract

Background: Monoclonal antibodies against calcitonin gene-related peptide (CGRP) or its receptor are efficacious for the prevention of migraine headaches. The downstream molecular mechanisms following ligand-receptor blockade by which these antibodies prevent CGRP signaling through CGRP receptors have not been demonstrated.

Methods: Here we produced tool monoclonal functional antagonist antibodies against CGRP and its canonical receptor and developed a novel cellular model using fluorogen-activated protein technology that allows detection of CGRP receptor internalization by flow cytometry and, for an extended time course, visualization by confocal microscopy.

Results: Using this cell model we showed that these antagonist antibodies block both CGRP-induced cAMP signaling and CGRP receptor internalization. At least 10-fold higher concentrations of either antibody are necessary to block CGRP receptor internalization compared with cAMP accumulation in our cell model.

Conclusion: These data reinforce our understanding of how monoclonal functional antagonist antibodies interfere with CGRP signaling.

Citing Articles

Mode and site of action of therapies targeting CGRP signaling.

Labastida-Ramirez A, Caronna E, Gollion C, Stanyer E, Dapkute A, Braniste D J Headache Pain. 2023; 24(1):125.

PMID: 37691118 PMC: 10494408. DOI: 10.1186/s10194-023-01644-8.

Pharmacological characterisation of erenumab, Aimovig, at two calcitonin gene-related peptide responsive receptors.

Garelja M, Alexander T, Bennie A, Nimick M, Petersen J, Walker C Br J Pharmacol. 2023; 181(1):142-161.

PMID: 37580864 PMC: 10840612. DOI: 10.1111/bph.16218.

CGRP physiology, pharmacology, and therapeutic targets: migraine and beyond.

Russo A, Hay D Physiol Rev. 2022; 103(2):1565-1644.

PMID: 36454715 PMC: 9988538. DOI: 10.1152/physrev.00059.2021.

Calcitonin gene-related peptide (CGRP) is a key molecule released in acute migraine attacks-Successful translation of basic science to clinical practice.

Edvinsson L J Intern Med. 2022; 292(4):575-586.

PMID: 35532284 PMC: 9546117. DOI: 10.1111/joim.13506.

The Arrival of Anti-CGRP Monoclonal Antibodies in Migraine.

Cohen F, Yuan H, DePoy E, Silberstein S Neurotherapeutics. 2022; 19(3):922-930.

PMID: 35426060 PMC: 9294119. DOI: 10.1007/s13311-022-01230-x.

References

Houle S, Larrivee J, Bachvarova M, BOUTHILLIER J, Bachvarov D, MARCEAU F . Antagonist-induced intracellular sequestration of rabbit bradykinin B(2) receptor. Hypertension. 2000; 35(6):1319-25. DOI: 10.1161/01.hyp.35.6.1319. View

Kuwasako K, Shimekake Y, Masuda M, Nakahara K, Yoshida T, Kitaura M . Visualization of the calcitonin receptor-like receptor and its receptor activity-modifying proteins during internalization and recycling. J Biol Chem. 2000; 275(38):29602-9. DOI: 10.1074/jbc.M004534200. View

Edvinsson L, Sams A, Jansen-Olesen I, Tajti J, Kane S, Rutledge R . Characterisation of the effects of a non-peptide CGRP receptor antagonist in SK-N-MC cells and isolated human cerebral arteries. Eur J Pharmacol. 2001; 415(1):39-44. DOI: 10.1016/s0014-2999(00)00934-1. View

Kearney J, Radbruch A, Liesegang B, Rajewsky K . A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 1979; 123(4):1548-50. View

Williamson D, Hill R, Shepheard S, Hargreaves R . The anti-migraine 5-HT(1B/1D) agonist rizatriptan inhibits neurogenic dural vasodilation in anaesthetized guinea-pigs. Br J Pharmacol. 2001; 133(7):1029-34. PMC: 1572868. DOI: 10.1038/sj.bjp.0704162. View

Gray J, Sheffler D, Bhatnagar A, Woods J, Hufeisen S, Benovic J . Cell-type specific effects of endocytosis inhibitors on 5-hydroxytryptamine(2A) receptor desensitization and resensitization reveal an arrestin-, GRK2-, and GRK5-independent mode of regulation in human embryonic kidney 293 cells. Mol Pharmacol. 2001; 60(5):1020-30. DOI: 10.1124/mol.60.5.1020. View

Kohler G, Milstein C . Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975; 256(5517):495-7. DOI: 10.1038/256495a0. View

Poyner D, Sexton P, Marshall I, Smith D, Quirion R, Born W . International Union of Pharmacology. XXXII. The mammalian calcitonin gene-related peptides, adrenomedullin, amylin, and calcitonin receptors. Pharmacol Rev. 2002; 54(2):233-46. DOI: 10.1124/pr.54.2.233. View

Silberstein S, Goadsby P . Migraine: preventive treatment. Cephalalgia. 2002; 22(7):491-512. DOI: 10.1046/j.1468-2982.2002.00386.x. View

10.

Pheng L, Dumont Y, Fournier A, Chabot J, Beaudet A, Quirion R . Agonist- and antagonist-induced sequestration/internalization of neuropeptide Y Y1 receptors in HEK293 cells. Br J Pharmacol. 2003; 139(4):695-704. PMC: 1573902. DOI: 10.1038/sj.bjp.0705306. View

11.

Cottrell G, Roosterman D, Marvizon J, Song B, Wick E, Pikios S . Localization of calcitonin receptor-like receptor and receptor activity modifying protein 1 in enteric neurons, dorsal root ganglia, and the spinal cord of the rat. J Comp Neurol. 2005; 490(3):239-55. DOI: 10.1002/cne.20669. View

12.

Cottrell G, Padilla B, Pikios S, Roosterman D, Steinhoff M, Grady E . Post-endocytic sorting of calcitonin receptor-like receptor and receptor activity-modifying protein 1. J Biol Chem. 2007; 282(16):12260-71. DOI: 10.1074/jbc.M606338200. View

13.

Padilla B, Cottrell G, Roosterman D, Pikios S, Muller L, Steinhoff M . Endothelin-converting enzyme-1 regulates endosomal sorting of calcitonin receptor-like receptor and beta-arrestins. J Cell Biol. 2007; 179(5):981-97. PMC: 2099187. DOI: 10.1083/jcb.200704053. View

14.

Chan K, Edvinsson L, Eftekhari S, Kimblad P, Kane S, Lynch J . Characterization of the calcitonin gene-related peptide receptor antagonist telcagepant (MK-0974) in human isolated coronary arteries. J Pharmacol Exp Ther. 2010; 334(3):746-52. DOI: 10.1124/jpet.110.165993. View

15.

Steiner T, Stovner L, Birbeck G . Migraine: the seventh disabler. J Headache Pain. 2013; 14:1. PMC: 3606966. DOI: 10.1186/1129-2377-14-1. View

16.

Vilardaga J, Jean-Alphonse F, Gardella T . Endosomal generation of cAMP in GPCR signaling. Nat Chem Biol. 2014; 10(9):700-6. PMC: 4417940. DOI: 10.1038/nchembio.1611. View

17.

Russell F, KING R, Smillie S, Kodji X, Brain S . Calcitonin gene-related peptide: physiology and pathophysiology. Physiol Rev. 2014; 94(4):1099-142. PMC: 4187032. DOI: 10.1152/physrev.00034.2013. View

18.

Bruchez M . Dark dyes-bright complexes: fluorogenic protein labeling. Curr Opin Chem Biol. 2015; 27:18-23. PMC: 4553107. DOI: 10.1016/j.cbpa.2015.05.014. View

19.

Pratt C, He J, Wang Y, Barth A, Bruchez M . Fluorogenic Green-Inside Red-Outside (GIRO) Labeling Approach Reveals Adenylyl Cyclase-Dependent Control of BKα Surface Expression. Bioconjug Chem. 2015; 26(9):1963-71. PMC: 4576318. DOI: 10.1021/acs.bioconjchem.5b00409. View

20.

Miller S, Liu H, Warfvinge K, Shi L, Dovlatyan M, Xu C . Immunohistochemical localization of the calcitonin gene-related peptide binding site in the primate trigeminovascular system using functional antagonist antibodies. Neuroscience. 2016; 328:165-83. DOI: 10.1016/j.neuroscience.2016.04.046. View