Cuproptosis promotes inflammatory osteolysis via GYS1-mediated glycogen metabolism

Abstract

Copper, predominantly present in bones, plays a crucial role in bone formation. However, when copper homeostasis is disrupted, excessive copper can trigger harmful inflammation and a novel form of cell death known as cuproptosis. The impact of cuproptosis on bone metabolism remains unclear. In this study, we demonstrated that excessive copper acts as an aggravator in osteoclastogenesis and bone resorption. We observed that the expression levels of the copper importer SLC31A1 and dihydrolipoamide S-acetyltransferase (DLAT) were positively correlated with bone loss in both human chronic apical periodontitis (CAP) tissues and mouse CAP models. Untargeted metabolomics analysis and screening of glucose metabolism enzymes revealed that glycogen synthesis was inhibited during cuproptosis. Mechanistically, excessive copper hindered glycogen synthesis via glycogen synthase 1 (GYS1), which limited the availability of glycogenolysis-derived glucose-6-phosphate (G6P) flux into pentose phosphate pathway (PPP), and was unable to yield abundant NADPH to ensure high demand of glutathione (GSH) for macrophage survival. The inhibition of glycogen synthesis intensified cuproptosis and bone-resorption activity. Moreover, excessive copper bound to H3K27me3, which further epigenetically inhibited the gene transcription of GYS1, thereby affecting glycogen synthesis and exacerbating cuproptosis and bone resorption. Furthermore, the disruption of glycogen metabolism intensified cuproptosis and promoted inflammatory bone loss in vivo. Our finding highlighted the complex interplay among copper homeostasis, glycogen metabolism, and the osteo-immune system, suggesting new therapeutic strategies for managing inflammatory bone diseases and other copper accumulation-related conditions through the metabolic reprogramming of cells.

References

[1] Takayanagi H. Inflammatory bone destruction and osteoimmunology. J. Periodontal Res, 2005, 40: 287-293

[2] Mbalaviele G, Novack DV, Schett G, Teitelbaum SL. Inflammatory osteolysis: a conspiracy against bone. J. Clin. Investig, 2017, 127: 2030-2039

[3] Zaidi M. Skeletal remodeling in health and disease. Nat. Med, 2007, 13: 791-801

[4] Tsukasaki M, Takayanagi H. Osteoimmunology: evolving concepts in bone-immune interactions in health and disease. Nat. Rev. Immunol., 2019, 19: 626-642

[5] Yahara Yet al.. Erythromyeloid progenitors give rise to a population of osteoclasts that contribute to bone homeostasis and repair. Nat. Cell Biol., 2020, 22: 49-59

[6] Okamoto Ket al.. Osteoimmunology: the conceptual framework unifying the immune and skeletal systems. Physiol. Rev., 2017, 97: 1295-1349

[7] Park JH, Lee NK, Lee SY. Current understanding of RANK signaling in osteoclast differentiation and maturation. Mol. Cells, 2017, 40: 706-713

[8] Kim JH, Kim N. Signaling pathways in osteoclast differentiation. Chonnam Med J., 2016, 52: 12-17

[9] Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature, 2003, 423: 337-342

[10] Han J, Luo J, Wang C, Kapilevich L, Zhang XA. Roles and mechanisms of copper homeostasis and cuproptosis in osteoarticular diseases. Biomed. Pharmacother., 2024, 174 116570

[11] Rondanelli, M. et al. Copper as dietary supplement for bone metabolism: a review. Nutrients, 13. (2021).

[12] Ciosek Z., Kot K., Rotter I. Iron, zinc, copper, cadmium, mercury, and bone tissue. Int. J. Environ. Res. Public Health, 20 (2023).

[13] Tsvetkov Pet al.. Copper induces cell death by targeting lipoylated TCA cycle proteins. Science, 2022, 375: 1254-1261

[14] Kahlson M, Dixon S. Copper-induced cell death. Science, 2022, 375: 1231-1232

[15] Sun Let al.. Lactylation of METTL16 promotes cuproptosis via mA-modification on FDX1 mRNA in gastric cancer. Nat. Commun., 2023, 14 6523

[16] Zhou L, Mao HQ, Wen YH, Chen Z, Zhang L. Cuproptosis Aggravates Pulpitis by Inhibiting the Pentose Phosphate Pathway. J. Dent. Res., 2025, 104: 541-550

[17] Jiang Zet al.. Hypoxia, cuproptosis, and osteoarthritis: unraveling the molecular crosstalk. Redox Biol., 2025, 85 103757

[18] Xiong C, Ling H, Hao Q, Zhou X. Cuproptosis: p53-regulated metabolic cell death?. Cell Death Differ., 2023, 30: 876-884

[19] Mao C, Wang M, Zhuang L, Gan B. Metabolic cell death in cancer: ferroptosis, cuproptosis, disulfidptosis, and beyond. Protein Cell, 2024, 15: 642-660

[20] Zhu SYet al.. COX17 restricts renal fibrosis development by maintaining mitochondrial copper homeostasis and restoring complex IV activity. Acta Pharm. Sin., 2023, 44: 2091-2102

[21] Ramchandani Det al.. Copper depletion modulates mitochondrial oxidative phosphorylation to impair triple negative breast cancer metastasis. Nat. Commun., 2021, 12 7311

[22] Caino MC, Altieri DC. Molecular pathways: mitochondrial reprogramming in tumor progression and therapy. Clin. Cancer Res., 2016, 22: 540-545

[23] Zhou L., Mao H. Q., Wen Y. H., Chen Z., Zhang L. Cuproptosis aggravates pulpitis by inhibiting the pentose phosphate pathway. J. Dent. Res. 220345251313797. (2025).

[24] Zhang H, Ma J, Tang K, Huang B. Beyond energy storage: roles of glycogen metabolism in health and disease. FEBS J., 2021, 288: 3772-3783

[25] Ma Jet al.. Glycogen metabolism regulates macrophage-mediated acute inflammatory responses. Nat. Commun., 2020, 11 1769

[26] Tang Ket al.. Hypoxia promotes breast cancer cell growth by activating a glycogen metabolic program. Cancer Res, 2021, 81: 4949-4963

[27] Chen Jet al.. Hepatic glycogenesis antagonizes lipogenesis by blocking S1P via UDPG. Science, 2024, 383 eadi3332

[28] Ma Ret al.. A Pck1-directed glycogen metabolic program regulates formation and maintenance of memory CD8(+) T cells. Nat. Cell Biol., 2018, 20: 21-27

[29] Mao HQet al.. STING inhibition alleviates bone resorption in apical periodontitis. Int. Endod. J., 2024, 57: 951-965

[30] Tsvetkov Pet al.. Mitochondrial metabolism promotes adaptation to proteotoxic stress. Nat. Chem. Biol., 2019, 15: 681-689

[31] Jiang P, Du W, Wu M. Regulation of the pentose phosphate pathway in cancer. Protein Cell, 2014, 5: 592-602

[32] Ren Jet al.. Glutathione ameliorates the meiotic defects of copper-exposed ovine oocytes via inhibiting the mitochondrial dysfunctions. Ecotoxicol. Environ. Saf., 2023, 251 114530

[33] Hodgkinson V, Petris MJ. Copper homeostasis at the host-pathogen interface. J. Biol. Chem., 2012, 287: 13549-13555

[34] Murdoch CC, Skaar EP. Nutritional immunity: the battle for nutrient metals at the host-pathogen interface. Nat. Rev. Microbiol, 2022, 20: 657-670

[35] Bandmann O, Weiss KH, Kaler SG. Wilson’s disease and other neurological copper disorders. Lancet Neurol., 2015, 14: 103-113

[36] Chen L, Min J, Wang F. Copper homeostasis and cuproptosis in health and disease. Signal Transduct. Target Ther., 2022, 7: 378

[37] Solier Set al.. A druggable copper-signalling pathway that drives inflammation. Nature, 2023, 617: 386-394

[38] Favaro Eet al.. Glucose utilization via glycogen phosphorylase sustains proliferation and prevents premature senescence in cancer cells. Cell Metab., 2012, 16: 751-764

[39] Zhang Het al.. TCR activation directly stimulates PYGB-dependent glycogenolysis to fuel the early recall response in CD8(+) memory T cells. Mol. Cell, 2022, 82: 3077-3088 e3076

[40] Liu Qet al.. Glycogen accumulation and phase separation drives liver tumor initiation. Cell, 2021, 184: 5559-5576 e5519

[41] Keinan Oet al.. Glycogen metabolism links glucose homeostasis to thermogenesis in adipocytes. Nature, 2021, 599: 296-301

[42] Sadiku Pet al.. Neutrophils fuel effective immune responses through gluconeogenesis and glycogenesis. Cell Metab., 2021, 33: 411-423.e414

[43] Xie Het al.. Glycogen metabolism is dispensable for tumour progression in clear cell renal cell carcinoma. Nat. Metab., 2021, 3: 327-336

[44] Roach PJ, Depaoli-Roach AA, Hurley TD, Tagliabracci VS. Glycogen and its metabolism: some new developments and old themes. Biochem. J., 2012, 441: 763-787

[45] Xie Xet al.. IKK/NF-kappaB and ROS signal axes are involved in Tenacissoside H mediated inhibitory effects on LPS-induced inflammatory osteolysis. Cell Prolif., 2024, 57 e13535

[46] Takahashi N, Udagawa N, Tanaka S, Suda T. Generating murine osteoclasts from bone marrow. Methods Mol. Med., 2003, 80: 129-144

[47] Lee NKet al.. A crucial role for reactive oxygen species in RANKL-induced osteoclast differentiation. Blood, 2005, 106: 852-859

[48] Lu Jet al.. Copper regulates the host innate immune response against bacterial infection via activation of ALPK1 kinase. Proc. Natl. Acad. Sci. USA, 2024, 121 e2311630121

[49] Chen SLet al.. GYS1 induces glycogen accumulation and promotes tumor progression via the NF-kappaB pathway in clear cell renal carcinoma. Theranostics, 2020, 10: 9186-9199

[50] Kim BE, Nevitt T, Thiele DJ. Mechanisms for copper acquisition, distribution and regulation. Nat. Chem. Biol., 2008, 4: 176-185

[51] Sun L, Zhang H, Gao P. Metabolic reprogramming and epigenetic modifications on the path to cancer. Protein Cell, 2022, 13: 877-919

[52] Su Het al.. Glutathione synthesis primes monocytes metabolic and epigenetic pathway for beta-glucan-trained immunity. Redox Biol., 2021, 48 102206

[53] Bannister AJ, Kouzarides T. Regulation of chromatin by histone modifications. Cell Res., 2011, 21: 381-395

[54] Millan-Zambrano G, Burton A, Bannister AJ, Schneider R. Histone post-translational modifications—cause and consequence of genome function. Nat. Rev. Genet, 2022, 23: 563-580

[55] Liu YTet al.. Dysregulated Wnt/beta-catenin signaling confers resistance to cuproptosis in cancer cells. Cell Death Differ., 2024, 31: 1452-1466

[56] Sun Met al.. Cuproptosis-related lncRNA JPX regulates malignant cell behavior and epithelial-immune interaction in head and neck squamous cell carcinoma via miR-193b-3p/PLAU axis. Int. J. Oral. Sci., 2024, 16: 63

Funding

National Natural Science Foundation of China (National Science Foundation of China)(82370948)

RIGHTS & PERMISSIONS

The Author(s)