title: Immunohistochemical localization of the calcitonin gene-related peptide binding site in the primate trigeminovascular system using functional antagonist antibodies.
abstract: Calcitonin gene-related peptide (CGRP) is a potent vasodilator and a neuromodulator implicated in the pathophysiology of migraine. It binds to the extracellular domains of calcitonin receptor-like receptor (CLR) and receptor activity-modifying protein (RAMP) 1 that together form the CGRP receptor. Antagonist antibodies against CGRP and its binding site at the receptor are clinically effective in preventing migraine attacks. The blood-brain barrier penetration of these antagonist antibodies is limited, suggesting that a potential peripheral site of action is sufficient to prevent migraine attacks. To further understand the sites of CGRP-mediated signaling in migraine, we used immunohistochemical staining with recently developed antagonist antibodies specifically recognizing a fusion protein of the extracellular domains of RAMP1 and CLR that comprise the CGRP binding pocket at the CGRP receptor in monkey and man. We confirmed binding of the antagonist antibodies to human vascular smooth muscle cells (VSMCs) of dural meningeal arteries and neurons in the trigeminal ganglion, both of which are likely sites of action for therapeutic antibodies in migraine patients. We further used one of these antibodies for detailed mapping on cynomolgus monkey tissue and found antagonist antibody binding sites at multiple levels in the trigeminovascular system: in the dura mater VSMCs, in neurons and satellite glial cells in the trigeminal ganglion, and in neurons in the spinal trigeminal nucleus caudalis. These data reinforce and clarify our understanding of CGRP receptor localization in a pattern consistent with a role for CGRP receptors in trigeminal sensitization and migraine pathology.
authors.affiliation: Department of Neuroscience, Amgen Inc., One Amgen Center Drive, Thousand Oaks, California 91320 and 360 Binney Street, Cambridge, MA 02142, USA. Electronic address: silkem@amgen.com.
authors.forename: Silke
authors.initials: S
authors.lastname: Miller
authors.affiliation: Department of Neuroscience, Amgen Inc., One Amgen Center Drive, Thousand Oaks, California 91320 and 360 Binney Street, Cambridge, MA 02142, USA.
authors.forename: Hantao
authors.initials: H
authors.lastname: Liu
authors.affiliation: University of Lund, Institute of Clinical Sciences at Lund University Hospital, House A13, Sölvegatan, Lund 22184, Sweden.
authors.forename: Karin
authors.initials: K
authors.lastname: Warfvinge
authors.affiliation: Department of Neuroscience, Amgen Inc., One Amgen Center Drive, Thousand Oaks, California 91320 and 360 Binney Street, Cambridge, MA 02142, USA.
authors.forename: Licheng
authors.initials: L
authors.lastname: Shi
authors.affiliation: Department of Neuroscience, Amgen Inc., One Amgen Center Drive, Thousand Oaks, California 91320 and 360 Binney Street, Cambridge, MA 02142, USA.
authors.forename: Mary
authors.initials: M
authors.lastname: Dovlatyan
authors.affiliation: Department of Neuroscience, Amgen Inc., One Amgen Center Drive, Thousand Oaks, California 91320 and 360 Binney Street, Cambridge, MA 02142, USA.
authors.forename: Cen
authors.initials: C
authors.lastname: Xu
authors.affiliation: University of Lund, Institute of Clinical Sciences at Lund University Hospital, House A13, Sölvegatan, Lund 22184, Sweden.
authors.forename: Lars
authors.initials: L
authors.lastname: Edvinsson
mesh_terms: D000368:Aged; D000818:Animals; D000906:Antibodies; D001665:Binding Sites; D015153:Blotting, Western; D058286:Calcitonin Receptor-Like Protein / Q000276:immunology / Q000378:metabolism; D045744:Cell Line, Tumor; D004388:Dura Mater / Q000098:blood supply / Q000166:cytology / Q000378:metabolism; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008252:Macaca fascicularis; D008297:Male; D008576:Meningeal Arteries / Q000166:cytology / Q000378:metabolism*; D008875:Middle Aged; D008881:Migraine Disorders / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D058262:Receptor Activity-Modifying Protein 1 / Q000276:immunology / Q000378:metabolism; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000276:immunology / Q000378:metabolism*; D012668:Trigeminal Ganglion / Q000166:cytology / Q000378:metabolism*
chemical_list: D000906:Antibodies; D058286:Calcitonin Receptor-Like Protein; C545502:RAMP1 protein, human; D058262:Receptor Activity-Modifying Protein 1; D018015:Receptors, Calcitonin Gene-Related Peptide
keywords: antibody; cynomolgus monkey; human; immunohistochemistry; migraine; trigeminal system