TY - JOUR
T1 - Bone pain induced by multiple myeloma is reduced by targeting V-ATPase and ASIC3
AU - Hiasa, Masahiro
AU - Okui, Tatsuo
AU - Allette, Yohance M.
AU - Ripsch, Matthew S.
AU - Sun-Wada, Ge Hong
AU - Wakabayashi, Hiroki
AU - Roodman, G. David
AU - White, Fletcher A.
AU - Yoneda, Toshiyuki
N1 - Funding Information:
This study was supported by the Project Development Team within the ICTSI NIH/NCRR (#TR000006), the Indiana University Health Strategic Research Initiative in Oncology, and start-up fund of Indiana University School of Medicine (T. Yoneda) and Merit Review Funds from the Veterans Administration (G.D. Roodman), the NIH (#DK100905 to F.A. White), MERIT Review Award (#BX002209) from the U.S. Department of Veterans Affairs, Biomedical Laboratory Research and Development Service (F.A. White), grants from St. Vincent Indianapolis Hospital and the St. Vincent Foundation to F.A. White. Support for Y.M. Allette as an Indiana CTSI Predoctoral trainee was provided by UL1 (#TR001108), NIH/NCATS (A. Shekhar, principal investigator), and Japan Society for the Promotion of Science Grants-in-aid for Research Activity Start-up, and Postdoctoral Fellowship for Research Abroad (M. Hiasa). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Publisher Copyright:
© 2017 AACR.
PY - 2017/3/15
Y1 - 2017/3/15
N2 - 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.
AB - 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.
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U2 - 10.1158/0008-5472.CAN-15-3545
DO - 10.1158/0008-5472.CAN-15-3545
M3 - Article
C2 - 28254863
AN - SCOPUS:85015822503
VL - 77
SP - 1283
EP - 1295
JO - Cancer Research
JF - Cancer Research
SN - 0008-5472
IS - 6
ER -