Abstract
The precise regulation of bone homeostasis and the balance between bone resorption and formation in periodontitis remain unclear. This study explores the role of long intergenic noncoding RNA-erythroid prosurvival (lincRNA-EPS) in inflammatory osteoclastogenesis and bone resorption. LincRNA-EPS knockout (KO) worsened LPS-induced alveolar bone resorption in vivo and osteoclast differentiation in vitro. Transcriptomics and protein sequencing showed dysregulated osteoclastogenesis and iron homeostasis without lincRNA-EPS, marked by increased expression of Lcn2. Knockdown of Lcn2 in osteoclast precursors (OCPs) resulted in a reduction in the level of iron metabolism and osteoclastogenesis; however, the regulatory response was delayed in KO cells. Correspondingly, overexpression of lincRNA-EPS accelerated the regulation of iron metabolism. Further, reducing Lcn2 levels in wildtype mice alleviated periodontitis-related bone loss, but not in KO mice. Taken together, we identified the critical role of lincRNA-EPS in regulating osteoclastogenesis under inflammatory environment, by preventing excessive iron metabolism caused by Lcn2.
Data availability
The mRNA sequencing datasets generated during the current study have been deposited in the NCBI Gene Expression Omnibus (GEO) database (GSE305332). The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (https://proteomecentral.proteomexchange.org) via the iProX partner repository with the dataset identifier PXD071554. Other data in this study are available from the corresponding author on reasonable request.
References
Tonetti MS, Greenwell H, Kornman KS. Staging and grading of periodontitis: framework and proposal of a new classification and case definition. J Periodontol. 2018;89:S159–72.
Sirisereephap K, Maekawa T, Tamura H, Hiyoshi T, Domon H, Isono T, et al. Osteoimmunology in periodontitis: local proteins and compounds to alleviate periodontitis. Int J Mol Sci. 2022;23:5540.
Usui M, Onizuka S, Sato T, Kokabu S, Ariyoshi W, Nakashima K. Mechanism of alveolar bone destruction in periodontitis—periodontal bacteria and inflammation. Jpn Dent Sci Rev. 2021;57:201–8.
Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature. 2003;423:337–42.
El-Masri BM, Andreasen CM, Laursen KS, Kofod VB, Dahl XG, Nielsen MH, et al. Mapping RANKL- and OPG-expressing cells in bone tissue: the bone surface cells as activators of osteoclastogenesis and promoters of the denosumab rebound effect. Bone Res. 2024;12:62.
Feng W, Guo J, Li M. RANKL-independent modulation of osteoclastogenesis. J Oral Biosci. 2019;61:16–21.
AlQranei MS, Chellaiah MA. Osteoclastogenesis in periodontal diseases: possible mediators and mechanisms. J Oral Biosci. 2020;62:123–30.
Xia Y, Inoue K, Du Y, Baker SJ, Reddy EP, Greenblatt MB, et al. TGFβ reprograms TNF stimulation of macrophages towards a non-canonical pathway driving inflammatory osteoclastogenesis. Nat Commun. 2022;13:3920.
Atianand MK, Hu W, Satpathy AT, Shen Y, Ricci EP, Alvarez-Dominguez JR, et al. A Long Noncoding RNA lincRNA-EPS acts as a transcriptional brake to restrain inflammation. Cell. 2016;165:1672–85.
Wu W, Hu A, Xu H, Su J. LincRNA-EPS alleviates inflammation in TMJ osteoarthritis by binding to SRSF3. J Dent Res. 2023;102:1141–51.
Hu A, Xiao F, Wu W, Xu H, Su J. LincRNA-EPS inhibits caspase-11 and NLRP3 inflammasomes in gingival fibroblasts to alleviate periodontal inflammation. Cell Prolif. 2024;57:e13539.
Guo H, Guo X, Jiang S. Long non-coding RNA lincRNA-erythroid prosurvival (EPS) alleviates cerebral ischemia/reperfusion injury by maintaining high-temperature requirement protein A1 (Htra1) stability through recruiting heterogeneous nuclear ribonucleoprotein L (HNRNPL). Bioengineered. 2022;13:12248–60.
Chen S, Zhu J, Sun L-Q, Liu S, Zhang T, Jin Y, et al. LincRNA-EPS alleviates severe acute pancreatitis by suppressing HMGB1-triggered inflammation in pancreatic macrophages. Immunology. 2021;163:201–19.
Zhang X, Wang Y, Su J. Effect of lincRNA-EPS on osteoclastogenesis of RAW264.7 cells induced by RANKL and LPS: an experimental study. J Oral Maxillofac Surg. 2024;34:163–9.
Hascoët E, Blanchard F, Blin-Wakkach C, Guicheux J, Lesclous P, Cloitre A. New insights into inflammatory osteoclast precursors as therapeutic targets for rheumatoid arthritis and periodontitis. Bone Res. 2023;11:26.
Hansen MS, Madsen K, Price M, Søe K, Omata Y, Zaiss MM, et al. Transcriptional reprogramming during human osteoclast differentiation identifies regulators of osteoclast activity. Bone Res. 2024;12:5.
Rohatgi N, Zou W, Li Y, Cho K, Collins PL, Tycksen E, et al. BAP1 promotes osteoclast function by metabolic reprogramming. Nat Commun. 2023;14:5923.
Li Y, Radu A-M, Rezaei A, Turner J, Shakib K, Obata A, et al. Si inhibited osteoclastogenesis: the role of Fe and the Fenton reaction. Adv Health Mater. 2025;14:e2501086.
Flo TH, Smith KD, Sato S, Rodriguez DJ, Holmes MA, Strong RK, et al. Lipocalin 2 mediates an innate immune response to bacterial infection by sequestrating iron. Nature. 2004;432:917–21.
Yang J, Goetz D, Li J-Y, Wang W, Mori K, Setlik D, et al. An iron delivery pathway mediated by a lipocalin. Mol Cell. 2002;10:1045–56.
Zhou J, Liu Y, Wei X, Yuan M, Zhang X, Qin L, et al. Glycnsisitin A: a promising bicyclic peptide against heart failure that facilitates TFRC-mediated uptake of iron in cardiomyocytes. Acta Pharm Sin B. 2024;14:3125–39.
Dowdle WE, Nyfeler B, Nagel J, Elling RA, Liu S, Triantafellow E, et al. Selective VPS34 inhibitor blocks autophagy and uncovers a role for NCOA4 in ferritin degradation and iron homeostasis in vivo. Nat Cell Biol. 2014;16:1069–79.
Knutson MD, Oukka M, Koss LM, Aydemir F, Wessling-Resnick M. Iron release from macrophages after erythrophagocytosis is up-regulated by ferroportin 1 overexpression and down-regulated by hepcidin. Proc Natl Acad Sci USA. 2005;102:1324–8.
Hempstead PD, Yewdall SJ, Fernie AR, Lawson DM, Artymiuk PJ, Rice DW, et al. Comparison of the three-dimensional structures of recombinant human H and horse L ferritins at high resolution. J Mol Biol. 1997;268:424–48.
Hentze MW, Muckenthaler MU, Andrews NC. Balancing acts: molecular control of mammalian iron metabolism. Cell. 2004;117:285–97.
Alam MI, Mae M, Farhana F, Oohira M, Yamashita Y, Ozaki Y, et al. NLRP3 inflammasome negatively regulates RANKL-induced osteoclastogenesis of mouse bone marrow macrophages but positively regulates it in the presence of lipopolysaccharides. Int J Mol Sci. 2022;23:6096.
Mbalaviele G, Novack DV, Schett G, Teitelbaum SL. Inflammatory osteolysis: a conspiracy against bone. J Clin Invest. 2017;127:2030–9.
Torres HM, Arnold KM, Oviedo M, Westendorf JJ, Weaver SR. Inflammatory processes affecting bone health and repair. Curr Osteoporos Rep. 2023;21:842–53.
Guo H-H, Xiong L, Pan J-X, Lee D, Liu K, Ren X, et al. Hepcidin contributes to Swedish mutant APP-induced osteoclastogenesis and trabecular bone loss. Bone Res. 2021;9:31.
Zhang J, Zhao H, Yao G, Qiao P, Li L, Wu S. Therapeutic potential of iron chelators on osteoporosis and their cellular mechanisms. Biomed Pharmacother. 2021;137:111380.
Zhang G, Zhen C, Yang J, Zhang Z, Wu Y, Che J, et al. 1-2 T static magnetic field combined with Ferumoxytol prevent unloading-induced bone loss by regulating iron metabolism in osteoclastogenesis. J Orthop Transl. 2023;38:126–40.
Devireddy LR, Gazin C, Zhu X, Green MR. A cell-surface receptor for lipocalin 24p3 selectively mediates apoptosis and iron uptake. Cell. 2005;123:1293–305.
Liu J, Song X, Kuang F, Zhang Q, Xie Y, Kang R, et al. NUPR1 is a critical repressor of ferroptosis. Nat Commun. 2021;12:647.
Srinivasan G, Aitken JD, Zhang B, Carvalho FA, Chassaing B, Shashidharamurthy R, et al. Lipocalin 2 deficiency dysregulates iron homeostasis and exacerbates endotoxin-induced sepsis. J Immunol. 2012;189:1911–9.
Lan Y, Yang T, Yue Q, Wang Z, Zhong X, Luo X, et al. IRP1 mediated ferroptosis reverses temozolomide resistance in glioblastoma via affecting LCN2/FPN1 signaling axis depended on NFKB2. iScience. 2023;26:107377.
Xia Y, Ge G, Xiao H, Wu M, Wang T, Gu C, et al. REPIN1 regulates iron metabolism and osteoblast apoptosis in osteoporosis. Cell Death Dis. 2023;14:631.
Wang D, Li X, Jiao D, Cai Y, Qian L, Shen Y, et al. LCN2 secreted by tissue-infiltrating neutrophils induces the ferroptosis and wasting of adipose and muscle tissues in lung cancer cachexia. J Hematol Oncol. 2023;16:30.
Capulli M, Ponzetti M, Maurizi A, Gemini-Piperni S, Berger T, Mak TW, et al. A complex role for lipocalin 2 in bone metabolism: global ablation in mice induces osteopenia caused by an altered energy metabolism. J Bone Min Res. 2018;33:1141–53.
Costa D, Lazzarini E, Canciani B, Giuliani A, Spanò R, Marozzi K, et al. Altered bone development and turnover in transgenic mice over-expressing lipocalin-2 in bone. J Cell Physiol. 2013;228:2210–21.
Kim H-J, Yoon H-J, Yoon K-A, Gwon M-R, Jin Seong S, Suk K, et al. Lipocalin-2 inhibits osteoclast formation by suppressing the proliferation and differentiation of osteoclast lineage cells. Exp Cell Res Exp Cell Res. 2015;334:301–9.
Yeom J, Ma S, Yim DJ, Lim Y-H. Surface proteins of Propionibacterium freudenreichii MJ2 inhibit RANKL-induced osteoclast differentiation by lipocalin-2 upregulation and lipocalin-2-mediated NFATc1 inhibition. Sci Rep. 2023;13:15644.
Galy B, Conrad M, Muckenthaler M. Mechanisms controlling cellular and systemic iron homeostasis. Nat Rev Mol Cell Biol. 2024;25:133–55.
Zhang DD. Thirty years of NRF2: advances and therapeutic challenges. Nat Rev Drug Discov. 2025;24:421–44.
Jeney V. Clinical impact and cellular mechanisms of iron overload-associated bone loss. Front Pharm. 2017;8:77.
Yang J, Li Q, Feng Y, Zeng Y. Iron deficiency and iron deficiency anemia: potential risk factors in bone loss. Int J Mol Sci. 2023;24:6891.
Agidigbi TS, Kim C. Reactive oxygen species in osteoclast differentiation and possible pharmaceutical targets of ROS-mediated osteoclast diseases. Int J Mol Sci. 2019;20:3576.
Lin L, Wang S, Deng H, Yang W, Rao L, Tian R, et al. Endogenous labile iron pool-mediated free radical generation for cancer chemodynamic therapy. J Am Chem Soc. 2020;142:15320–30.
Fibach E, Prus E. The labile iron pool in normal and pathological erythroid cells - analysis by flow cytometry. Blood. 2005;106:3597.
Pan W, Wang Q, Chen Q. The cytokine network involved in the host immune response to periodontitis. Int J Oral Sci. 2019;11:30.
Zhang X, Hartmann P. How to calculate sample size in animal and human studies. Front Med (Lausanne). 2023;10:1215927.
Acknowledgements
This work was supported by the National Natural Science Foundation of China (82201077).
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This work was supported by the National Natural Science Foundation of China (82201077).
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JW, YW and JS performed study concept and design; JW and YW performed acquisition, analysis and interpretation of data, statistical analysis, writing, review and revision of the paper; ZZ and XW provided technical and material support. All authors read and approved the final paper.
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Ethical approval of animal experiments was obtained from the Ethics Review Board of the Affiliated Stomatology Hospital of Tongji University (NO. [2022]-DW-29) and were performed in compliance with ARRIVE guidelines.
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Wang, J., Wang, Y., Zhang, Z. et al. LincRNA-EPS alleviates osteoclastogenesis under inflammatory microenvironment through preventing excessive iron metabolism. Cell Death Dis (2026). https://doi.org/10.1038/s41419-026-08716-y
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DOI: https://doi.org/10.1038/s41419-026-08716-y
