A humanized eNAMPT -neutralizing monoclonal antibody reduces cancer-induced bone pain and osteoclastogenesis in murine breast cancer model
- 1Department of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ, United States
- 2The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology, University of Florida, Jupiter, FL, United States
Abstract
Introduction: Cancer-induced bone pain (CIBP) is a common symptom among patients with metastasized cancer, however, health professionals often face challenges in managing patients’ pain. Extracellular nicotinamide phosphoribosyltransferase (eNAMPT) is a DAMP overexpressed in metastatic breast carcinoma that binds to Toll-Like Receptor 4 (TLR4) to potentially trigger a signaling cascade through Nuclear Factor kappa β (NFk-β) inducing inflammation, osteoclastogenesis, and contributing to CIBP. To assess the impact of eNAMPT on CIBP and to identify alternative therapeutic options, ALT-100, an eNAMPT-neutralizing monoclonal antibody, was tested in murine and cell-based models.
Methods: First, eNAMPT synthesis and secretion by EO771 breast adenocarcinoma cells and its modulation by ALT-100 were assessed using eNAMPT ELISA. Next, recombinant eNAMPT-induced osteoclastogenesis and its inhibition by ALT-100 were evaluated in RAW 264.7 osteoclast precursor cells using TRAP staining. In vivo, female C57BL/6J mice underwent intramedullary femoral injection of EO771 cells to establish CIBP or SHAM surgery. One week later, mice received ALT-100 (4 mg/kg, ip) or saline control, followed by longitudinal assessment of pain-related behaviors and femoral radiographs.
Results: The eNAMPT ELISA assay revealed that EO771 cells synthesize and secrete eNAMPT in a time-dependent manner maximal at 48 h, which were reduced by ALT-100 mAb. In RAW 264.7 cells, recombinant eNAMPT significantly increased TRAP activity, while co-administration of ALT-100 significantly reduced this effect. In the murine CIBP model, ALT-100 significantly reduced pain-related behaviors compared to saline control mice in the cancer group. Moreover, radiographic analysis demonstrated that ALT-100 mitigates cancer-induced bone loss.
Discussion: These data show that ALT-100 exhibits antinociceptive effects in a CIBP murine model and inhibits eNAMPT-induced osteoclast differentiation. Our results indicate that blockade of eNAMPT can potentially serve as an alternative clinical strategy for treating CIBP.
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BibTeX
@article{mathur2026humanized,
title = {{A humanized eNAMPT -neutralizing monoclonal antibody reduces cancer-induced bone pain and osteoclastogenesis in murine breast cancer model}},
author = {Nikhil C. Mathur and Ethan Villaroel and Isabella Villaroel and Erika Liktor-Busa and Kelly L. Karlage and Manvir Kaur and Carrie Kempf and Beatrice Gudaite and Zuo Ding and Joe G. N. Garcia and Tally M. Largent-Milnes and Todd W. Vanderah},
journal = {Frontiers in Pain Research},
year = {2026},
month = oct,
doi = {10.3389/fpain.2026.1955262},
url = {https://doi.org/10.3389/fpain.2026.1955262},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - A humanized eNAMPT -neutralizing monoclonal antibody reduces cancer-induced bone pain and osteoclastogenesis in murine breast cancer model
AU - Nikhil C. Mathur
AU - Ethan Villaroel
AU - Isabella Villaroel
AU - Erika Liktor-Busa
AU - Kelly L. Karlage
AU - Manvir Kaur
AU - Carrie Kempf
AU - Beatrice Gudaite
AU - Zuo Ding
AU - Joe G. N. Garcia
AU - Tally M. Largent-Milnes
AU - Todd W. Vanderah
PY - 2026
DA - 2026/10/21
JO - Frontiers in Pain Research
DO - 10.3389/fpain.2026.1955262
UR - https://doi.org/10.3389/fpain.2026.1955262
AB - Introduction: Cancer-induced bone pain (CIBP) is a common symptom among patients with metastasized cancer, however, health professionals often face challenges in managing patients’ pain. Extracellular nicotinamide phosphoribosyltransferase (eNAMPT) is a DAMP overexpressed in metastatic breast carcinoma that binds to Toll-Like Receptor 4 (TLR4) to potentially trigger a signaling cascade through Nuclear Factor kappa β (NFk-β) inducing inflammation, osteoclastogenesis, and contributing to CIBP. To assess the impact of eNAMPT on CIBP and to identify alternative therapeutic options, ALT-100, an eNAMPT-neutralizing monoclonal antibody, was tested in murine and cell-based models. Methods: First, eNAMPT synthesis and secretion by EO771 breast adenocarcinoma cells and its modulation by ALT-100 were assessed using eNAMPT ELISA. Next, recombinant eNAMPT-induced osteoclastogenesis and its inhibition by ALT-100 were evaluated in RAW 264.7 osteoclast precursor cells using TRAP staining. In vivo, female C57BL/6J mice underwent intramedullary femoral injection of EO771 cells to establish CIBP or SHAM surgery. One week later, mice received ALT-100 (4 mg/kg, ip) or saline control, followed by longitudinal assessment of pain-related behaviors and femoral radiographs. Results: The eNAMPT ELISA assay revealed that EO771 cells synthesize and secrete eNAMPT in a time-dependent manner maximal at 48 h, which were reduced by ALT-100 mAb. In RAW 264.7 cells, recombinant eNAMPT significantly increased TRAP activity, while co-administration of ALT-100 significantly reduced this effect. In the murine CIBP model, ALT-100 significantly reduced pain-related behaviors compared to saline control mice in the cancer group. Moreover, radiographic analysis demonstrated that ALT-100 mitigates cancer-induced bone loss. Discussion: These data show that ALT-100 exhibits antinociceptive effects in a CIBP murine model and inhibits eNAMPT-induced osteoclast differentiation. Our results indicate that blockade of eNAMPT can potentially serve as an alternative clinical strategy for treating CIBP.
SN - 2673-561X
ER -