Targeting ferroptosis suppresses osteocyte glucolipotoxicity and alleviates diabetic osteoporosis

Abstract

Diabetic osteoporosis (DOP) is the leading complication continuously threatening the bone health of patients with diabetes. A key pathogenic factor in DOP is loss of osteocyte viability. However, the mechanism of osteocyte death remains unclear. Here, we identified ferroptosis, which is iron-dependent programmed cell death, as a critical mechanism of osteocyte death in murine models of DOP. The diabetic microenvironment significantly enhanced osteocyte ferroptosis in vitro, as shown by the substantial lipid peroxidation, iron overload, and aberrant activation of the ferroptosis pathway. RNA sequencing showed that heme oxygenase-1 (HO-1) expression was notably upregulated in ferroptotic osteocytes. Further findings revealed that HO-1 was essential for osteocyte ferroptosis in DOP and that its promoter activity was controlled by the interaction between the upstream NRF2 and c-JUN transcription factors. Targeting ferroptosis or HO-1 efficiently rescued osteocyte death in DOP by disrupting the vicious cycle between lipid peroxidation and HO-1 activation, eventually ameliorating trabecular deterioration. Our study provides insight into DOP pathogenesis, and our results provide a mechanism-based strategy for clinical DOP treatment.

References

[1] Napoli N et al. Mechanisms of diabetes mellitus-induced bone fragility. Nat. Rev. Endocrinol., 2017, 13: 208-219

[2] Johnell O, Kanis JA. An estimate of the worldwide prevalence and disability associated with osteoporotic fractures. Osteoporos. Int., 2006, 17: 1726-1733

[3] Shanbhogue VV, Hansen S, Frost M, Brixen K, Hermann AP. Bone disease in diabetes: another manifestation of microvascular disease? Lancet Diabetes Endocrinol., 2017, 5: 827-838

[4] Kim JH et al. Tumor necrosis factor-α antagonist diminishes osteocytic RANKL and sclerostin expression in diabetes rats with periodontitis. PLoS One, 2017, 12: e0189702

[5] Lai X et al. The dependences of osteocyte network on bone compartment, age, and disease. Bone Res., 2015, 3: 15009

[6] Dallas SL, Prideaux M, Bonewald LF. The osteocyte: an endocrine cell… and more. Endocr. Rev., 2013, 34: 658-690

[7] He Z et al. Irisin inhibits osteocyte apoptosis by activating the Erk signaling pathway in vitro and attenuates ALCT-induced osteoarthritis in mice. Bone, 2020, 141: 115573

[8] Eckhardt BA et al. Accelerated osteocyte senescence and skeletal fragility in mice with type 2 diabetes. JCI Insight, 2020, 5: 9

[9] Yang L et al. High glucose downregulates connexin 43 expression and its gap junction and hemichannel function in osteocyte-like MLO-Y4 cells through activation of the p38MAPK/ERK signal pathway. Diabetes Metab. Syndr. Obes., 2020, 13: 545-557

[10] Yao X et al. Chondrocyte ferroptosis contribute to the progression of osteoarthritis. J. Orthop. Translat., 2021, 27: 33-43

[11] Hadian K, Stockwell BR. SnapShot: ferroptosis. Cell, 2020, 181: 1188-1188.e1

[12] Lee H et al. Energy-stress-mediated AMPK activation inhibits ferroptosis. Nat. Cell Biol., 2020, 22: 225-234

[13] Miotto G et al. Insight into the mechanism of ferroptosis inhibition by ferrostatin-1. Redox Biol., 2020, 28: 101328

[14] Fang X et al. Ferroptosis as a target for protection against cardiomyopathy. Proc. Natl. Acad. Sci. USA, 2019, 116: 2672-2680

[15] Maher P, Currais A, Schubert D. Using the oxytosis/ferroptosis pathway to understand and treat age-associated neurodegenerative diseases. Cell Chem. Biol., 2020, 27: 1456-1471

[16] Liang C, Zhang X, Yang M, Dong X. Recent progress in ferroptosis inducers for cancer therapy. Adv. Mater., 2019, 31: e1904197

[17] Cui W, Liu D, Gu W, Chu B. Peroxisome-driven ether-linked phospholipids biosynthesis is essential for ferroptosis. Cell Death Differ., 2021, 28: 2536-2551

[18] Zhou Y et al. Verification of ferroptosis and pyroptosis and identification of PTGS2 as the hub gene in human coronary artery atherosclerosis. Free Radic. Biol. Med., 2021, 171: 55-68

[19] Yang WS et al. Regulation of ferroptotic cancer cell death by GPX4. Cell, 2014, 156: 317-331

[20] Al Saedi A, Myers DE, Stupka N, Duque G. 1,25(OH)D ameliorates palmitate-induced lipotoxicity in human primary osteoblasts leading to improved viability and function. Bone, 2020, 141: 115672

[21] Ma H et al. Melatonin suppresses ferroptosis induced by high glucose via activation of the Nrf2/HO-1 signaling pathway in type 2 diabetic osteoporosis. Oxid. Med. Cell Longev., 2020, 2020: 9067610

[22] Chiang SK, Chen SE, Chang LC. A dual role of heme oxygenase-1 in cancer cells. Int. J. Mol. Sci., 2018, 20: 39

[23] Loboda A, Damulewicz M, Pyza E, Jozkowicz A, Dulak J. Role of Nrf2/HO-1 system in development, oxidative stress response and diseases: an evolutionarily conserved mechanism. Cell Mol. Life Sci., 2016, 73: 3221-3247

[24] Yang CC et al. Induction of HO-1 by 5, 8-dihydroxy-4’,7-dimethoxyflavone via activation of ROS/p38 MAPK/Nrf2 attenuates thrombin-induced connective tissue growth factor expression in human cardiac fibroblasts. Oxid. Med. Cell Longev., 2020, 2020: 1080168

[25] Bellezza I, Giambanco I, Minelli A, Donato R. Nrf2-Keap1 signaling in oxidative and reductive stress. Biochim. Biophys. Acta Mol. Cell Res., 2018, 1865: 721-733

[26] Yoo EJ et al. TonEBP suppresses the HO-1 gene by blocking recruitment of Nrf2 to its promoter. Front. Immunol., 2019, 10: 850

[27] Zhang Q et al. DUB3 deubiquitinates and stabilizes NRF2 in chemotherapy resistance of colorectal cancer. Cell Death Differ., 2019, 26: 2300-2313

[28] Takamoto I, Kadowaki T. Diabetes and osteoporosis. Clin. Calcium, 2004, 14: 255-261

[29] Zhang WL, Meng HZ, Yang MW. Regulation of DMT1 on bone microstructure in type 2 diabetes. Int. J. Med. Sci., 2015, 12: 441-449

[30] Botolin S, McCabe LR. Bone loss and increased bone adiposity in spontaneous and pharmacologically induced diabetic mice. Endocrinology, 2007, 148: 198-205

[31] Motyl K, McCabe LR. Streptozotocin, type I diabetes severity and bone. Biol. Proced. Online, 2009, 11: 296-315

[32] Takanche JS, Kim JE, Han SH, Yi HK. Effect of gomisin A on osteoblast differentiation in high glucose-mediated oxidative stress. Phytomedicine, 2020, 66: 153107

[33] Kim S et al. Osteoblastic glucocorticoid signaling exacerbates high-fat-diet-induced bone loss and obesity. Bone Res., 2021, 9: 40

[34] An Y et al. Activation of ROS/MAPKs/NF-κB/NLRP3 and inhibition of efferocytosis in osteoclast-mediated diabetic osteoporosis. FASEB J., 2019, 33: 12515-12527

[35] Andreev D et al. Osteocyte necrosis triggers osteoclast-mediated bone loss through macrophage-inducible C-type lectin. J. Clin. Investig., 2020, 130: 4811-4830

[36] McHugh J. Osteocyte death promotes bone loss. Nat. Rev. Rheumatol., 2020, 16: 539

[37] Han Y, You X, Xing W, Zhang Z, Zou W. Paracrine and endocrine actions of bone-the functions of secretory proteins from osteoblasts, osteocytes, and osteoclasts. Bone Res., 2018, 6: 16

[38] Maurel DB et al. Characterization of a novel murine Sost ER Cre model targeting osteocytes. Bone Res., 2019, 7: 6

[39] Li MCM, Chow SKH, Wong RMY, Qin L, Cheung WH. The role of osteocytes-specific molecular mechanism in regulation of mechanotransduction – a systematic review. J. Orthop. Translat., 2021, 29: 1-9

[40] Emerton KB et al. Osteocyte apoptosis and control of bone resorption following ovariectomy in mice. Bone, 2010, 46: 577-583

[41] Jiang T et al. Gastrodin protects against glutamate-induced ferroptosis in HT-22 cells through Nrf2/HO-1 signaling pathway. Toxicol Vitro, 2020, 62: 104715

[42] Adedoyin O et al. Heme oxygenase-1 mitigates ferroptosis in renal proximal tubule cells. Am. J. Physiol. Ren. Physiol., 2018, 314: F702-F714

[43] Tang Z et al. HO-1-mediated ferroptosis as a target for protection against retinal pigment epithelium degeneration - ScienceDirect. Redox Biol., 2021, 43: 101971

[44] Zhang C et al. Nuciferine ameliorates hepatic steatosis in high-fat diet/streptozocin-induced diabetic mice through a PPARα/PPARγ coactivator-1α pathway. Br. J. Pharmacol., 2018, 175: 4218-4228

[45] Wu X et al. Kindlin-2 deletion in osteoprogenitors causes severe chondrodysplasia and low-turnover osteopenia in mice. J. Orthop. Translat., 2022, 32: 41-48

[46] Xu G, Hu X, Han L, Zhao Y, Li Z. The construction of a novel xenograft bovine bone scaffold, (DSS)6-liposome/CKIP-1 siRNA/calcine bone and its osteogenesis evaluation on skull defect in rats. J. Orthop. Transl., 2021, 28: 74-82

[47] Hong D et al. Deletion of TMEM268 inhibits growth of gastric cancer cells by downregulating the ITGB4 signaling pathway. Cell Death Differ., 2019, 26: 1453-1466

[48] Cheung Tung Shing KS et al. EPO does not promote interaction between the erythropoietin and beta-common receptors. Sci. Rep., 2018, 8

[49] Li H et al. FOXP1 drives osteosarcoma development by repressing P21 and RB transcription downstream of P53. Oncogene, 2021, 40: 2785-2802

Funding

National Natural Science Foundation of China (National Science Foundation of China)(81802679)

China Postdoctoral Science Foundation(2018M632136)