m⁶A-modified DRAM1 recognized by YTHDF1 regulates autophagy during dexamethasone-induced osteogenic inhibition

Abstract

Bone health is critically dependent on the precise regulation of bone-forming cells, which are significantly impacted by glucocorticoids. Although DNA damage-regulated autophagy modulator 1 (DRAM1) plays a key role in autophagy, its involvement in glucocorticoid-induced osteoporosis has not been fully explored. Here, we showed that alterations in N⁶-methyladenosine (m⁶A) levels contributed to the regulation of DRAM1 under dexamethasone treatment. In vitro, our research revealed that high doses of dexamethasone impaired the m⁶A-dependent regulation of DRAM1 by YTH m⁶A RNA binding protein F1 (YTHDF1), leading to decreased DRAM1 levels and subsequent disruption of autophagy-associated osteogenic differentiation in human bone marrow mesenchymal stem cells (hBMSCs) and MC3T3-E1 cells. Additionally, the classic Wnt/β-catenin pathway, which plays a critical role in bone formation, was shown to be modulated by DRAM1 during osteogenic differentiation. In vivo experiments showed that DRAM1 protein expression in the femurs of Ythdf1 knockout (KO) mice was significantly lower than that in the wild-type (WT) group, and that resveratrol not only mitigated dexamethasone-induced bone damage but was also associated with increased DRAM1 expression. These findings advance our understanding of how glucocorticoids hinder bone formation and suggest that targeting the m⁶A-dependent YTHDF1/DRAM1 regulatory axis may offer novel strategies for osteoporosis treatment.

Keywords

autophagy / dexamethasone / N⁶-methyladenosine / osteogenic differentiation / Wnt/β-catenin signalling

References

[1] Liu J, You Y, Sun Z, et al. WTAP-mediated m6A RNA methylation regulates the differentiation of bone marrow mesenchymal stem cells via the miR-29b-3p/HDAC4 axis. Stem Cells Transl Med. 2023; 12: 307-321.

[2] Hu K, Adachi JD. Glucocorticoid induced osteoporosis. Expert Rev Endocrinol Metabolism. 2019; 14: 259-266.

[3] Compston J. Glucocorticoid-induced osteoporosis: an update. Endocrine. 2018; 61: 7-16.

[4] Van Staa TP, Leufkens HG, Abenhaim L, Zhang B, Cooper C. Use of oral corticosteroids and risk of fractures. J Bone Miner Res. 2000; 15: 993-1000.

[5] Yao W, Dai W, Jiang L, et al. Sclerostin-antibody treatment of glucocorticoid-induced osteoporosis maintained bone mass and strength. Osteop Int. 2016; 27: 283-294.

[6] Tang YH, Yue ZS, Li GS, et al. Effect of β-ecdysterone on glucocorticoid-induced apoptosis and autophagy in osteoblasts. Mol Med Rep. 2018; 17: 158-164.

[7] Chen L, Shi X, Weng S, et al. Vitamin K2 can rescue the dexamethasone-induced downregulation of osteoblast autophagy and mitophagy thereby restoring osteoblast function in vitro and in vivo. Front Pharmacol. 2020; 11: 1209.

[8] Yang X, Jiang T, Wang Y, Guo L. The role and mechanism of SIRT1 in resveratrol-regulated osteoblast autophagy in osteoporosis rats. Sci Rep. 2019; 9:18424.

[9] Huang J, Ye Y, Xiao Y, et al. Geniposide ameliorates glucocorticoid-induced osteoblast apoptosis by activating autophagy. Biomed Pharmacother. 2022; 155:113829.

[10] Zhang C, Wang S, Lu X, et al. POP1 facilitates proliferation in triple-negative breast cancer via m6A-dependent degradation of CDKN1A mRNA. Research (Washington, DC). 2024; 7: 0472.

[11] Ou X, Tan Y, Xie J, et al. Methylation of GPRC5A promotes liver metastasis and docetaxel resistance through activating mTOR signaling pathway in triple negative breast cancer. Drug Resist Updat. 2024; 73:101063.

[12] Shi D, Liu X, Li X, et al. Yth m(6)A RNA-binding protein 1 regulates osteogenesis of MC3T3-E1 cells under hypoxia via translational control of thrombospondin-1. Int J Mol Sci. 2023; 24(2): 1741.

[13] Yang S, Wei J, Cui Y, et al. m(6)A mRNA demethylase FTO regulates melanoma tumorigenicity and response to anti-PD-1 blockade. Nat Commun. 2019; 10: 2782.

[14] Yang Y, Hsu PJ, Chen YS, Yang YG. Dynamic transcriptomic m(6)A decoration: writers, erasers, readers and functions in RNA metabolism. Cell Res. 2018; 28: 616-624.

[15] Wang X, Zhao B, Roundtree I, et al. N(6)-methyladenosine modulates messenger RNA translation efficiency. Cell. 2015; 161: 1388-1399.

[16] Jiang X, Liu B, Nie Z, et al. The role of m6A modification in the biological functions and diseases. Signal Transduct Target Ther. 2021; 6: 74.

[17] Wang L, Zhu L, Liang C, et al. Targeting N6-methyladenosine reader YTHDF1 with siRNA boosts antitumor immunity in NASH-HCC by inhibiting EZH2-IL-6 axis. J Hepatol. 2023; 79: 1185-1200.

[18] Liu T, Zheng X, Wang C, et al. The m(6)A “reader” YTHDF1 promotes osteogenesis of bone marrow mesenchymal stem cells through translational control of ZNF839. Cell Death Dis. 2021; 12: 1078.

[19] van der Vaart M, Korbee CJ, Lamers GE, et al. The DNA damage-regulated autophagy modulator DRAM1 links mycobacterial recognition via TLR-MYD88 to autophagic defense. Cell Host Microbe. 2014; 15(6): 753-767.

[20] Nagata M, Arakawa S, Yamaguchi H, et al. Dram1 regulates DNA damage-induced alternative autophagy. Cell Stress. 2018; 2: 55-65.

[21] Lu T, Zhu Z, Wu J, et al. DRAM1 regulates autophagy and cell proliferation via inhibition of the phosphoinositide 3-kinase-Akt-mTOR-ribosomal protein S6 pathway. Cell Commun Signal. 2019; 17: 28.

[22] Yu M, Jiang Y, Feng Q, Ouyang Y, Gan J. DRAM1 protects neuroblastoma cells from oxygen-glucose deprivation/reperfusion-induced injury via autophagy. Int J Mol Sci. 2014; 15: 19253-19264.

[23] Zhang XD, Qi L, Wu JC, Qin ZH. DRAM1 regulates autophagy flux through lysosomes. PLoS One. 2013; 8:e63245.

[24] Tang N, Zhao H, Zhang H, Dong Y. Effect of autophagy gene DRAM on proliferation, cell cycle, apoptosis, and autophagy of osteoblast in osteoporosis rats. J Cell Physiol. 2019; 234: 5023-5032.

[25] Liu T, Li B, Zheng X, et al. Chordin-like 1 improves osteogenesis of bone marrow mesenchymal stem cells through enhancing BMP4-SMAD pathway. Front Endocrinol. 2019; 10: 360.

[26] Chung CY, Iida-Klein A, Wyatt LE, et al. Serial passage of MC3T3-E1 cells alters osteoblastic function and responsiveness to transforming growth factor-beta1 and bone morphogenetic protein-2. Biochem Biophys Res Commun. 1999; 265: 246-251.

[27] Zhu C, Zheng X, Yang Y, et al. LGR4 acts as a key receptor for R-spondin 2 to promote osteogenesis through Wnt signaling pathway. Cell Signal. 2016; 28: 989-1000.

[28] Gu Q, Chen M, Zhang Y, Huang Y, Yang H, Shi Q. Haem oxygenase-1 induction prevents glucocorticoid-induced osteoblast apoptosis through activation of extracellular signal-regulated kinase1/2 signalling pathway. J Orthop Translat. 2019; 19: 29-37.

[29] Wang J, Liu J, Guo Y. Cell growth stimulation, cell cycle alternation, and anti-apoptosis effects of bovine bone collagen hydrolysates derived peptides on MC3T3-E1 cells ex vivo. Molecules. 2020; 25(10): 2305.

[30] Sule R, Rivera G, Gomes AV. Western blotting (immunoblotting): history, theory, uses, protocol and problems. BioTechniques. 2023; 75: 99-114.

[31] Zhang S, Zhang N, Wan T, et al. Oncometabolite D-2HG drives tumor metastasis and protumoral macrophage polarization by targeting FTO/m(6)A/ANGPTL4/integrin axis in triple-negative breast cancer. J Exp Clin Cancer Res. 2025; 44: 41.

[32] Shi H, Zhang X, Weng Y, et al. m(6)A facilitates hippocampus-dependent learning and memory through YTHDF1. Nature. 2018; 563: 249-253.

[33] Ran Z, Wang Y, Li J, et al. 3D-printed biodegradable magnesium alloy scaffolds with zoledronic acid-loaded ceramic composite coating promote osteoporotic bone defect repair. Int J Bioprint. 2023; 9: 769.

[34] Jiang Y, Luo W, Wang B, Wang X, Gong P, Xiong Y. Resveratrol promotes osteogenesis via activating SIRT1/FoxO1 pathway in osteoporosis mice. Life Sci. 2020; 246:117422.

[35] Zhang X, Su T, Wu Y, et al. N6-methyladenosine reader YTHDF1 promotes stemness and therapeutic resistance in hepatocellular carcinoma by enhancing NOTCH1 expression. Cancer Res. 2024; 84: 827-840.

[36] Nazarian A, Snyder BD, Zurakowski D, Müller R. Quantitative micro-computed tomography: a non-invasive method to assess equivalent bone mineral density. Bone. 2008; 43: 302-311.

[37] Zheng H, Xu W, Zhou W, et al. Beraprost ameliorates postmenopausal osteoporosis by regulating Nedd4-induced Runx2 ubiquitination. Cell Death Dis. 2021; 12: 497.

[38] Crighton D, Wilkinson S, O'Prey J, et al. DRAM, a p53-induced modulator of autophagy, is critical for apoptosis. Cell. 2006; 126: 121-134.

[39] Blommaart EF, Luiken JJ, Blommaart PJ, van Woerkom GM, Meijer AJ. Phosphorylation of ribosomal protein S6 is inhibitory for autophagy in isolated rat hepatocytes. J Biol Chem. 1995; 270: 2320-2326.

[40] Yang Y, Sun Y, Mao W, Zhang H, Ni B, Jiang L. Oxidative stress induces downregulation of TP53INP2 and suppresses osteogenic differentiation of BMSCs during osteoporosis through the autophagy degradation pathway. Free Radical Biol Med. 2021; 166: 226-237.

[41] Wang L, Li Q, Yan H, et al. Resveratrol protects osteoblasts against dexamethasone-induced cytotoxicity through activation of AMP-activated protein kinase. Drug Des Dev Ther. 2020; 14: 4451-4463.

[42] Luo Q, Liu S, Xie L, et al. Resveratrol ameliorates glucocorticoid-induced bone damage in a zebrafish model. Front Pharmacol. 2019; 10: 195.

[43] Kao C, Tai L, Chiou S, et al. Resveratrol promotes osteogenic differentiation and protects against dexamethasone damage in murine induced pluripotent stem cells. Stem Cells Dev. 2010; 19: 247-258.

[44] Teitelbaum SL. Glucocorticoids and the osteoclast. Clin Exp Rheumatol. 2015; 33: S37-39.

[45] Suvorova II, Knyazeva AR, Pospelov VA. Resveratrol-induced p53 activation is associated with autophagy in mouse embryonic stem cells. Biochem Biophys Res Commun. 2018; 503: 2180-2185.

[46] Wan W, Ao X, Chen Q, et al. METTL3/IGF2BP3 axis inhibits tumor immune surveillance by upregulating N(6)-methyladenosine modification of PD-L1 mRNA in breast cancer. Mol Cancer. 2022; 21: 60.

[47] Shen G, Ren H, Shang Q, et al. Autophagy as a target for glucocorticoid-induced osteoporosis therapy. Cell Mol Life Sci. 2018; 75: 2683-2693.

[48] Zhang Y, Li M, Liu Z, Fu Q. Arbutin ameliorates glucocorticoid-induced osteoporosis through activating autophagy in osteoblasts. Exp Med (Maywood, NJ). 2021; 246: 1650-1659.

[49] Zeng C, Wang S, Gu H, et al. Galangin mitigates glucocorticoid-induced osteoporosis by activating autophagy of BMSCs via triggering the PKA/CREB signaling pathway. Acta Biochim Biophys Sin. 2023; 55: 1275-1287.

[50] Ma Q, Yu J, Zhang X, Wu X, Deng G. Wnt/β-catenin signaling pathway-a versatile player in apoptosis and autophagy. Biochimie. 2023; 211: 57-67.

[51] Rocha MA, de Campos Vidal B, Mello MLS. Sodium valproate modulates the methylation status of lysine residues 4, 9 and 27 in histone H3 of HeLa cells. Current Mol Pharmacol. 2023; 16: 197-210.

[52] Zinovkina LA, Makievskaya CI, Galkin II, Zinovkin RA. Mitochondria-targeted uncouplers decrease inflammatory reactions in endothelial cells by enhancing methylation of the ICAM1 gene promoter. Curr Mol Pharmacol. 2024; 17:e150823219723.

[53] Ullah A, Razzaq A, Alfaifi MY, et al. Sanguinarine attenuates lung cancer progression via oxidative stress-induced cell apoptosis. Curr Mol Pharmacol. 2024; 17:e18761429269383.

[54] Ziegler R, Kasperk C. Glucocorticoid-induced osteoporosis: prevention and treatment. Steroids. 1998; 63: 344-348.

[55] Xiao Z, Xiao X, Kong LB, et al. Preparation of MXene-based hybrids and their application in neuromorphic devices. Int J Extreme Manuf. 2024; 6.

[56] Arcos D, Boccaccini A, Bohner M, et al. The relevance of biomaterials to the prevention and treatment of osteoporosis. Acta Biomater. 2014; 10: 1793-1805.

[57] Lei C, Song J, Li S, et al. Advances in materials-based therapeutic strategies against osteoporosis. Biomaterials. 2023; 296:122066.

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2026 The Author(s). Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.