{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T05:18:31Z","timestamp":1787807911420,"version":"build-2784847793"},"reference-count":48,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T00:00:00Z","timestamp":1787788800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Osteoporosis remains a major global health challenge, and current single-pathway therapies often fail to achieve coordinated bone remodeling. Dickkopf-1 (DKK-1) is a potent inhibitor of the Wnt\/\u03b2-catenin signaling pathway and plays a critical role in osteoporotic bone loss. Neutralization of DKK-1 represents a promising therapeutic strategy to promote bone formation while simultaneously inhibiting bone resorption.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>A humanized anti-DKK-1 monoclonal antibody (IgG4) was generated via CDR grafting combined with structure-guided back-mutations. Its binding affinity and specificity were assessed by ELISA and surface plasmon resonance. The antibody's ability to restore Wnt\/\u03b2-catenin signaling was evaluated in human bone marrow mesenchymal stem cells (hMSCs) by Western blotting. Osteogenic differentiation and mineralization were assessed by ALP activity assay and Alizarin Red S staining, respectively. An osteoblast-osteoclast Transwell co-culture model was established to evaluate the antibody's effects on osteoclastogenesis via the OPG\/RANKL axis, with osteoclast differentiation assessed by TRAP staining and qPCR. Developability properties, including stability and immunogenicity, were also characterized.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The humanized antibody bound to DKK-1 with high affinity (EC\n                      <jats:sub>\u2085\u2080<\/jats:sub>\n                      = 32.8 ng\/mL, KD = 2.760 \u00d7 10\n                      <jats:sup>\u207b\u00b9\u2070<\/jats:sup>\n                      mol\/L) and showed no cross-reactivity to DKK-2, DKK-3, or DKK-4. In hMSCs, the antibody restored Wnt\/\u03b2-catenin signaling, as evidenced by nuclear \u03b2-catenin accumulation, GSK-3\u03b2 phosphorylation, and c-Myc upregulation, leading to increased ALP activity, OPG secretion, and mineralization. In the Transwell co-culture model, the antibody restored the OPG\/sRANKL molar ratio and significantly suppressed osteoclast marker gene expression (ACP5, CALCR, CTSK, ITGB3, MMP9, and NFATC1) as well as TRAP-positive multinucleated cell formation (from 29 \u00b1 5 per field in the undifferentiated control to 2 \u00b1 1 per field in the antibody-treated group). The antibody also exhibited favorable developability properties, including long-term stability at 2 -8\u00b0C for over two years and low cellular immunogenicity.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>These findings demonstrate that the humanized anti-DKK-1 antibody exerts dual anabolic and anti-resorptive effects through the Wnt\/\u03b2-catenin\/OPG\/RANKL axis. This dual mechanism of action positions the antibody as a promising therapeutic candidate for osteoporosis, warranting further preclinical and clinical evaluation.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2026.1906434","type":"journal-article","created":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T04:27:40Z","timestamp":1787804860000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["A dual-action humanized DKK-1 antibody that activates the Wnt\/\u03b2-catenin signaling pathway and inhibits the RANKL signaling pathway for osteoporosis therapy"],"prefix":"10.3389","volume":"17","author":[{"given":"Sheng","family":"Hou","sequence":"first","affiliation":[{"name":"Taizhou Mabtech Pharmaceuticals Co., Ltd","place":["Taizhou, China"]},{"name":"State key laboratory of macromolecular drugs and large-scale preparation, NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies, 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China"]},{"name":"State key laboratory of macromolecular drugs and large-scale preparation, School of Pharmaceutical Sciences, Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yimei","family":"Wu","sequence":"additional","affiliation":[{"name":"Taizhou Mabtech Pharmaceuticals Co., Ltd","place":["Taizhou, China"]},{"name":"State key laboratory of macromolecular drugs and large-scale preparation, NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies, Shanghai Zhangjiang Biotechnology Co., Ltd","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Wang","sequence":"additional","affiliation":[{"name":"Taizhou Mabtech Pharmaceuticals Co., Ltd","place":["Taizhou, China"]},{"name":"State key laboratory of macromolecular drugs and large-scale preparation, NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies, Shanghai 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Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Xu","sequence":"additional","affiliation":[{"name":"State key laboratory of macromolecular drugs and large-scale preparation, NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies, Shanghai Zhangjiang Biotechnology Co., Ltd","place":["Shanghai, China"]},{"name":"State key laboratory of macromolecular drugs and large-scale preparation, School of Pharmaceutical Sciences and Food Engineering, Liaocheng University","place":["Liaocheng, China"]},{"name":"State key laboratory of macromolecular drugs and large-scale preparation, School of Pharmaceutical Sciences, Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huaizu","family":"Guo","sequence":"additional","affiliation":[{"name":"State key laboratory of macromolecular drugs and large-scale preparation, NMPA Key Laboratory for 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