title: Bone Pain Induced by Multiple Myeloma Is Reduced by Targeting V-ATPase and ASIC3.
abstract: Multiple myeloma patients experience severe bone pain (MMBP) that is undertreated and poorly understood. In this study, we studied MMBP in an intratibial mouse xenograft model that employs JJN3 human multiple myeloma cells. In this model, mice develop MMBP associated in bone with increased sprouting of calcitonin gene-related peptide-positive (CGRP+) sensory nerves and in dorsal root ganglia (DRG) with upregulation of phosphorylated ERK1/2 (pERK1/2) and pCREB, two molecular indicators of neuron excitation. We found that JJN3 cells expressed a vacuolar proton pump (V-ATPase) that induced an acidic bone microenvironment. Inhibition of JJN3-colonized bone acidification by a single injection of the selective V-ATPase inhibitor, bafilomycin A1, decreased MMBP, CGRP+sensory neuron sprouting, and pERK1/2 and pCREB expression in DRG. CGRP+sensory nerves also expressed increased levels of the acid-sensing nociceptor ASIC3. Notably, a single injection of the selective ASIC3 antagonist APETx2 dramatically reduced MMBP in the model. Mechanistic investigations in primary DRG neurons cocultured with JJN3 cells showed increased neurite outgrowth and excitation inhibited by bafilomycin A1 or APETx2. Furthermore, combining APETx2 with bafilomycin A1 reduced MMBP to a greater extent than either agent alone. Finally, combining bafilomycin A1 with the osteoclast inhibitor zoledronic acid was sufficient to ameliorate MMBP, which was refractory to zoledronic acid. Overall, our results show that osteoclasts and multiple myeloma cooperate to induce an acidic bone microenvironment that evokes MMBP as a result of the excitation of ASIC3-activated sensory neurons. Furthermore, they present a mechanistic rationale for targeting ASIC3 on neurons along with the multiple myeloma-induced acidic bone microenvironment as a strategy to relieve MMBP in patients.Cancer Res; 77(6); 1283-95. ©2017 AACR.
authors.affiliation: Division of Hematology and Oncology, Indiana University School of Medicine, Indianapolis, Indiana.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Hiasa
authors.affiliation: Division of Hematology and Oncology, Indiana University School of Medicine, Indianapolis, Indiana.
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Okui
authors.affiliation: Department of Anesthesia, Paul and Carole Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, Indiana.
authors.forename: Yohance M
authors.initials: YM
authors.lastname: Allette
authors.affiliation: Department of Anesthesia, Paul and Carole Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, Indiana.
authors.forename: Matthew S
authors.initials: MS
authors.lastname: Ripsch
authors.affiliation: Department of Biochemistry, Faculty of Pharmaceutical Sciences, Doshisha Women's College, Kyoto, Japan.
authors.forename: Ge-Hong
authors.initials: GH
authors.lastname: Sun-Wada
authors.affiliation: Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, Mie, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Wakabayashi
authors.affiliation: Division of Hematology and Oncology, Indiana University School of Medicine, Indianapolis, Indiana.|The Roudebusch VA, Indianapolis, Indiana.
authors.forename: G David
authors.initials: GD
authors.lastname: Roodman
authors.affiliation: Department of Anesthesia, Paul and Carole Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, Indiana.
authors.forename: Fletcher A
authors.initials: FA
authors.lastname: White
authors.affiliation: Division of Hematology and Oncology, Indiana University School of Medicine, Indianapolis, Indiana. toshyone@iu.edu.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Yoneda
mesh_terms: D062565:Acid Sensing Ion Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000818:Animals; D050071:Bone Density Conservation Agents / Q000494:pharmacology; D001847:Bone Diseases / Q000209:etiology / Q000378:metabolism / Q000517:prevention & control*; D001862:Bone Resorption / Q000209:etiology / Q000378:metabolism / Q000517:prevention & control*; D002478:Cells, Cultured; D004164:Diphosphonates / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005727:Ganglia, Spinal / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D018942:Macrolides / Q000494:pharmacology; D051379:Mice; D016513:Mice, SCID; D009101:Multiple Myeloma / Q000150:complications*; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D010146:Pain / Q000209:etiology / Q000378:metabolism / Q000517:prevention & control*; D025262:Vacuolar Proton-Translocating ATPases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000077211:Zoledronic Acid
chemical_list: C508378:ASIC3 protein, mouse; D062565:Acid Sensing Ion Channels; D050071:Bone Density Conservation Agents; D004164:Diphosphonates; D004791:Enzyme Inhibitors; D007093:Imidazoles; D018942:Macrolides; D000077211:Zoledronic Acid; C040929:bafilomycin A1; D025262:Vacuolar Proton-Translocating ATPases