Original articles
Yingzi He, Zhiwei Zheng, Chang Liu, Wen Li, Liping Zhao, Guohui Nie, Huawei Li. Inhibiting DNA methylation alleviates cisplatin-induced hearing loss by decreasing oxidative stress-induced mitochondria-dependent apoptosis via the LRP1-PI3K/AKT pathway[J]. Acta Pharmaceutica Sinica B, 2022, 12(3): 1305-1321

Inhibiting DNA methylation alleviates cisplatin-induced hearing loss by decreasing oxidative stress-induced mitochondria-dependent apoptosis via the LRP1-PI3K/AKT pathway
Yingzi Hea,b, Zhiwei Zhenga,b, Chang Liua,b, Wen Lia,b, Liping Zhaoa,b, Guohui Niee, Huawei Lia,b,c,d
a. ENT Institute and Otorhinolaryngology Department of Eye & ENT Hospital, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200031, China;
b. NHC Key Laboratory of Hearing Medicine (Fudan University), Shanghai 200031, China;
c. Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China;
d. The Institutes of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China;
e. Department of Otolaryngology and Institute of Translational Medicine, Shenzhen Second People's Hospital/the First Affiliated Hospital of Shenzhen University Health Science Center, Shenzhen 518035, China
Cisplatin-related ototoxicity is a critical side effect of chemotherapy and can lead to irreversible hearing loss. This study aimed to assess the potential effect of the DNA methyltransferase (DNMT) inhibitor RG108 on cisplatin-induced ototoxicity. Immunohistochemistry, apoptosis assay, and auditory brainstem response (ABR) were employed to determine the impacts of RG108 on cisplatin-induced injury in murine hair cells (HCs) and spiral ganglion neurons (SGNs). Rhodamine 123 and TMRM were utilized for mitochondrial membrane potential (MMP) assessment. Reactive oxygen species (ROS) amounts were evaluated by Cellrox green and Mitosox-red probes. Mitochondrial respiratory function evaluation was performed by determining oxygen consumption rates (OCRs). The results showed that RG108 can markedly reduce cisplatin induced damage in HCs and SGNs, and alleviate apoptotic rate by protecting mitochondrial function through preventing ROS accumulation. Furthermore, RG108 upregulated BCL-2 and downregulated APAF1, BAX, and BAD in HEI-OC1 cells, and triggered the PI3K/AKT pathway. Decreased expression of low-density lipoprotein receptor-related protein 1 (LRP1) and high methylation of the LRP1 promoter were observed after cisplatin treatment. RG108 treatment can increase LRP1 expression and decrease LRP1 promoter methylation. In conclusion, RG108 might represent a new potential agent for preventing hearing loss induced by cisplatin via activating the LRP1-PI3K/AKT pathway.
Key words:    Cisplatin    DNMT    Apoptosis    Hair cell    Spiral ganglion neurons    RG108    Mitochondrial dysfunction    ROS   
Received: 2021-06-03     Revised: 2021-10-29
DOI: 10.1016/j.apsb.2021.11.002
Funds: The authors would like to thank Yalin Huang for help with the confocal microscope. This work was supported by grants from the National Key R&D Program of China (No. 2017YFA0103900), the National Natural Science Foundation of China (Nos. 82071045, 81870728, 81830029, and 81970875), and Shanghai Rising-Star Program (19QA1401800).
Corresponding author: Guohui Nie,E-mai:nieguohui@email.szu.edu.cn;Huawei Li,E-mai:hwli@shmu.edu.cn     Email:nieguohui@email.szu.edu.cn;hwli@shmu.edu.cn
Author description:
PDF(KB) Free
Yingzi He
Zhiwei Zheng
Chang Liu
Wen Li
Liping Zhao
Guohui Nie
Huawei Li

[1] Rybak LP, Whitworth CA, Mukherjea D, Rarakumar V. Mechanisms of cisplatin-induced ototoxicity and prevention. Hear Res 2007; 226: 157-167
[2] Karasawa T, Steyger PS. An integrated view of cisplatin-induced nephrotoxicity and ototoxicity. Toxicol Lett 2015; 237: 219-227
[3] Kim YJ, Kim J, Tian C, Lim HJ, Kim YS, Chung JH, et al. Prevention of cisplatin-induced ototoxicity by the inhibition of gap junctional intercellular communication in auditory cells. Cell Mol Life Sci 2014; 71: 3859-3871
[4] Rybak LP, Mukherjea D, Jajoo S, Ramkumar V. Cisplatin ototoxicity and protection: clinical and experimental studies. Tohoku J Exp Med 2009; 219: 177-186
[5] Mohan S, Smyth BJ, Namin A, Phillips G, Gratton MA. Targeted amelioration of cisplatin-induced ototoxicity in guinea pigs. Otolaryngol Head Neck Surg 2014; 151: 836-839
[6] Dickey DT, Muldoon LL, Doolittle ND, Peterson DR, Kraemer DF, Neuwelt EA. Effect of N-acetylcysteine route of administration on chemoprotection against cisplatin-induced toxicity in rat models. Cancer Chemother Pharmacol 2008; 62: 235-241
[7] Choe WT, Chinosornvatana N, Chang KW. Prevention of cisplatin ototoxicity using transtympanic N-acetylcysteine and lactate. Otol Neurotol 2004; 25: 910-915
[8] Wimmer C, Mees K, Stumpf T, Welsch U, Reichel O, Suckfull M. Round window application of D-methionine, sodium thiosulfate, brain-derived neurotrophic factor, and fibroblast growth factor-2 in cisplatin-induced ototoxicity. Otol Neurotol 2004; 25: 33-40
[9] Kouzarides T. Chromatin modifications and their function. Cell 2007; 128: 693-705
[10] Jones PA. Functions of DNA methylation: islands, start sites, gene bodies and beyond. Nat Rev Genet 2012; 13: 484-492
[11] Zheng BN, Ding CH, Chen SJ, Zhu K, Shao J, Feng J, et al. Targeting PRMT5 activity inhibits the malignancy of hepatocellular carcinoma by promoting the transcription of HNF4alpha. Theranostics 2019; 9: 2606-2617
[12] Wang P, Zhang P, Huang J, Li M, Chen X. Trichostatin A protects against cisplatin-induced ototoxicity by regulating expression of genes related to apoptosis and synaptic function. Neurotoxicology 2013; 37: 51-62
[13] Yu H, Lin Q, Wang Y, He Y, Fu S, Jiang H, et al. Inhibition of H3K9 methyltransferases G9a/GLP prevents ototoxicity and ongoing hair cell death. Cell Death Dis 2013; 4:e506
[14] He YZ, Li W, Zheng ZW, Zhao LP, Li WY, Wang YF, et al. Inhibition of protein arginine methyltransferase 6 reduces reactive oxygen species production and attenuates aminoglycoside- and cisplatin-induced hair cell death. Theranostics 2020; 10: 133-150
[15] Jones PA, Takai D. The role of DNA methylation in mammalian epigenetics. Science 2001; 293: 1068-1070
[16] Jaenisch R, Bird A. Epigenetic regulation of gene expression: how the genome integrates intrinsic and environmental signals. Nat Genet 2003; 33: 245-254
[17] Bird AP. Cpg-rich islands and the function of DNA methylation. Nature 1986; 321: 209-213
[18] Turek-Plewa J, Jagodzinski PP. The role of mammalian DNA methyltransferases in the regulation of gene expression. Cell Mol Biol Lett 2005; 10: 631-647
[19] Robertson KD. DNA methylation, methyltransferases, and cancer. Oncogene 2001; 20: 3139-3155
[20] Okano M, Bell DW, Haber DA, Li E. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell 1999; 99: 247-257
[21] Sanz LA, Kota SK, Feil R. Genome-wide DNA demethylation in mammals. Genome Biol 2010; 11:110
[22] Guo F, Li XL, Liang D, Li T, Zhu P, Guo HS, et al. Active and passive demethylation of male and female pronuclear DNA in the mammalian zygote. Cell Stem Cell 2014; 15: 447-458
[23] Zhou Y, Hu ZQ. Epigenetic DNA demethylation causes inner ear stem cell differentiation into hair cell-like cells. Front Cell Neurosci 2016; 10:185
[24] Zhou Y, Hu ZQ. Genome-wide demethylation by 5-aza-2'-deoxycytidine alters the cell fate of stem/progenitor cells. Stem Cell Rev Rep 2015; 11: 87-95
[25] Deng X, Liu ZJ, Li XY, Zhou Y, Hu ZQ. Generation of new hair cells by DNA methyltransferase (Dnmt) inhibitor 5-azacytidine in a chemically-deafened mouse model. Sci Rep 2019; 9:7997
[26] Fahy J, Jeltsch A, Arimondo PB. DNA methyltransferase inhibitors in cancer: a chemical and therapeutic patent overview and selected clinical studies. Expert Opin Ther Pat 2012; 22: 1427-1442
[27] Siedlecki P, Boy RG, Musch T, Brueckner B, Suhai S, Lyko F, et al. Discovery of two novel, small-molecule inhibitors of DNA methylation. J Med Chem 2006; 49: 678-683
[28] Stresemann C, Lyko F. Modes of action of the DNA methyltransferase inhibitors azacytidine and decitabine. Int J Cancer 2008; 123: 8-13
[29] Tokarz P, Kaarniranta K, Blasiak J. Inhibition of DNA methyltransferase or histone deacetylase protects retinal pigment epithelial cells from DNA damage induced by oxidative stress by the stimulation of antioxidant enzymes. Eur J Pharmacol 2016; 776: 167-175
[30] Teitz T, Fang J, Goktug AN, Bonga JD, Diao SY, Hazlitt RA, et al. CDK2 inhibitors as candidate therapeutics for cisplatin-and noise-induced hearing loss. J Exp Med 2018; 215: 1187-1203
[31] Fears CY, Grammer JR, Stewart JE, Jr, Annis DS, Mosher DF, Bornstein P, et al. Low-density lipoprotein receptor-related protein contributes to the antiangiogenic activity of thrombospondin-2 in a murine glioma model. Cancer Res 2005; 65: 9338-9346
[32] Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 2014; 30: 2114-2120
[33] Dehne N, Lautermann J, Petrat F, Rauen U, de Groot H. Cisplatin ototoxicity: involvement of iron and enhanced formation of superoxide anion radicals. Toxicol Appl Pharm 2001; 174: 27-34
[34] Wu M, Neilson A, Swift AL, Moran R, Tamagnine J, Parslow D, et al. Multiparameter metabolic analysis reveals a close link between attenuated mitochondrial bioenergetic function and enhanced glycolysis dependency in human tumor cells. Am J Physiol Cell Physiol 2007; 292: C125-C136
[35] Zheng ZW, Wang YF, Yu HQ, Li W, Wu JF, Cai CF, et al. Salvianolic acid B inhibits ototoxic drug-induced ototoxicity by suppression of the mitochondrial apoptosis pathway. J Cell Mol Med 2020; 24: 6883-6897
[36] Chuang TY, Guo Y, Seki SM, Rosen AM, Johanson DM, Mandell JW, et al. LRP1 expression in microglia is protective during CNS autoimmunity. Acta Neuropathol Commun 2016; 4:68
[37] Peng JH, Pang JW, Huang L, Enkhjargal B, Zhang TY, Mo J, et al. LRP1 activation attenuates white matter injury by modulating microglial polarization through Shc1/PI3K/Akt pathway after subarachnoid hemorrhage in rats. Redox Biol 2019; 21:101121
[38] Ruggiero A, Trombatore G, Triarico S, Arena R, Ferrara P, Scalzone M, et al. Platinum compounds in children with cancer: toxicity and clinical management. Anticancer Drugs 2013; 24: 1007-1019
[39] Chen FQ, Schacht J, Sha SH. Aminoglycoside-induced histone deacetylation and hair cell death in the mouse cochlea. J Neurochem 2009; 108: 1226-1236
[40] Drottar M, Liberman MC, Ratan RR, Roberson DW. The histone deacetylase inhibitor sodium butyrate protects against cisplatin-induced hearing loss in guinea pigs. Laryngoscope 2006; 116: 292-296
[41] Yang DH, Xie J, Liu K, Peng Z, Guo JY, Yu SK, et al. The histone deacetylase inhibitor sodium butyrate protects against noise-induced hearing loss in Guinea pigs. Neurosci Lett 2017; 660: 140-146
[42] Uysal F, Akkoyunlu G, Ozturk S. Dynamic expression of DNA methyltransferases (DNMTs) in oocytes and early embryos. Biochimie 2015; 116: 103-113
[43] Yan F, Shen N, Pang JX, Zhao N, Zhang YW, Bode AM, et al. A vicious loop of fatty acid-binding protein 4 and DNA methyltransferase 1 promotes acute myeloid leukemia and acts as a therapeutic target. Leukemia 2018; 32: 865-873
[44] Chestnut BA, Chang Q, Price A, Lesuisse C, Wong M, Martin LJ. Epigenetic regulation of motor neuron cell death through DNA methylation. J Neurosci 2011; 31: 16619-16636
[45] Borse V, Al Aameri RFH, Sheehan K, Sheth S, Kaur T, Mukherjea D, et al. Epigallocatechin-3-gallate, a prototypic chemopreventative agent for protection against cisplatin-based ototoxicity. Cell Death Dis 2017; 8:e2921
[46] Ghosh S, Sheth S, Sheehan K, Mukherjea D, Dhukhwa A, Borse V, et al. The endocannabinoid/cannabinoid receptor 2 system protects against cisplatin-induced hearing loss. Front Cell Neurosci 2018; 12:271
[47] Kujawa SG, Liberman MC. Adding insult to injury: cochlear nerve degeneration after "“Temporary" ” noise-induced hearing loss. J Neurosci 2009; 29: 14077-14085
[48] Sheth S, Mukherjea D, Rybak LP, Ramkumar V. Mechanisms of cisplatin-induced ototoxicity and otoprotection. Front Cell Neurosci 2017; 11:338
[49] Schacht J, Talaska AE, Rybak LP. Cisplatin and aminoglycoside antibiotics: hearing loss and its prevention. Anat Rec 2012; 295: 1837-1850
[50] Rybak LP, Kelly T. Ototoxicity: bioprotective mechanisms. Curr Opin Otolaryngol Head Neck Surg 2003; 11: 328-333
[51] Rybak LP, Mukherjea D, Ramkumar V. Mechanisms of cisplatin-induced ototoxicity and prevention. Semin Hear 2019; 40: 197-204
[52] Tokunaga E, Oki E, Egashira A, Sadanaga N, Morita M, Kakeji Y, et al. Deregulation of the Akt pathway in human cancer. Curr Cancer Drug Targets 2008; 8: 27-36
[53] Kennedy SG, Wagner AJ, Conzen SD, Jordan J, Bellacosa A, Tsichlis PN, et al. The PI3-kinase/Akt signaling pathway delivers an anti-apoptotic signal. Gene Dev 1997; 11: 701-713
[54] Orike N, Middleton G, Borthwick E, Buchman V, Cowen T, Davies AM. Role of PI3-kinase, Akt and Bcl-2-related proteins in sustaining the survival of neurotrophic factor-independent adult sympathetic neurons. J Cell Biol 2001; 154: 995-1005
[55] Aburto MR, Magarinos M, Leon Y, Varela-Nieto I, Sanchez-Calderon H. AKT signaling mediates IGF-I survival actions on otic neural progenitors. PLoS One 2012; 7:e30790
[56] Brand Y, Levano S, Radojevic V, Naldi AM, Setz C, Ryan AF, et al. All Akt isoforms (Akt1, Akt2, Akt3) are involved in normal hearing, but only Akt2 and Akt3 are involved in auditory hair cell survival in the mammalian inner ear. PLoS One 2015; 10:e0121599
[57] Selivanova O, Brieger J, Heinrich UR, Mann W. Akt and c-Jun N-terminal kinase are regulated in response to moderate noise exposure in the cochlea of guinea pigs. ORL J Otorhinolaryngol Relat Spec 2007; 69: 277-282
[58] Chung WH, Pak K, Lin B, Webster N, Ryan AF. A PI3K pathway mediates hair cell survival and opposes gentamicin toxicity in neonatal rat organ of Corti. J Assoc Res Otolaryngol 2006; 7: 373-382
[59] Haake SM, Dinh CT, Chen S, Eshraghi AA, Van De Water TR. Dexamethasone protects auditory hair cells against TNFalpha-initiated apoptosis via activation of PI3K/Akt and NFkappaB signaling. Hear Res 2009; 255: 22-32
[60] Kucharava K, Sekulic-Jablanovic M, Horvath L, Bodmer D, Petkovic V. Pasireotide protects mammalian cochlear hair cells from gentamicin ototoxicity by activating the PI3K-Akt pathway. Cell Death Dis 2019; 10:110
[61] Fernandez-Castaneda A, Arandjelovic S, Stiles TL, Schlobach RK, Mowen KA, Gonias SL, et al. Identification of the low density lipoprotein (LDL) receptor-related protein-1 interactome in central nervous system myelin suggests a role in the clearance of necrotic cell debris. J Biol Chem 2013; 288: 4538-4548
[62] Gardai SJ, McPhillips KA, Frasch SC, Janssen WJ, Starefeldt A, Murphy-Ullrich JE, et al. Cell-surface calreticulin initiates clearance of viable or apoptotic cells through trans-activation of LRP on the phagocyte. Cell 2005; 123: 321-334
[63] Campana WM, Li XQ, Dragojlovic N, Janes J, Gaultier A, Gonias SL. The low-density lipoprotein receptor-related protein is a pro-survival receptor in Schwann cells: possible implications in peripheral nerve injury. J Neurosci 2006; 26: 11197-11207
[64] Zurhove K, Nakajima C, Herz J, Bock HH, May P. Gamma-secretase limits the inflammatory response through the processing of LRP1. Sci Signal 2008; 1:ra15
Similar articles:
1.Benyu Nan, Zirui Zhao, Kanglun Jiang, Xi Gu, Huawei Li, Xinsheng Huang.Astaxanthine attenuates cisplatin ototoxicity in vitro and protects against cisplatin-induced hearing loss in vivo[J]. Acta Pharmaceutica Sinica B, 2022,12(1): 167-181
2.Layla Shojaie, Myra Ali, Andrea Iorga, Lily Dara.Mechanisms of immune checkpoint inhibitor-mediated liver injury[J]. Acta Pharmaceutica Sinica B, 2021,11(12): 3727-3739
3.Jiyu Zhou, Ningning Huang, Yitong Guo, Shuang Cui, Chaoliang Ge, Qingxian He, Xiaojie Pan, Guangji Wang, Hong Wang, Haiping Hao.Combined obeticholic acid and apoptosis inhibitor treatment alleviates liver fibrosis[J]. Acta Pharmaceutica Sinica B, 2019,9(3): 526-536
4.Guangyao Lv, Dengjun Sun, Jingwen Zhang, Xiaoxia Xie, Xiaoqiong Wu, Weishuo Fang, Jingwei Tian, Chunhong Yan, Hongbo Wang, Fenghua Fu.Lx2-32c, a novel semi-synthetic taxane, exerts antitumor activity against prostate cancer cells in vitro and in vivo[J]. Acta Pharmaceutica Sinica B, 2017,7(1): 52-58
Similar articles: