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Histone Deacetylase Inhibitor RGFP109 Overcomes Temozolomide Resistance by Blocking NF-κB-Dependent Transcription in Glioblastoma Cell Lines

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Abstract

Glioblastoma (GBM) is the most frequent and aggressive tumour in the central nervous system. Many studies have demonstrated that upregulation of the NF-κB onco-pathway is accompanied by the acquisition of Temozolomide (TMZ) resistance in GBM cells. Here, we show that RGFP109, a selective histone deacetylase (HDAC1 and HDAC3) inhibitor, overcomes TMZ resistance and downregulates the expression of NF-κB-regulated pro-survival genes in a TMZ-resistant (TR) GBM cell line. RGFP109 did not alter the phosphorylation levels of NF-κB/p65 or inhibitory κBα (IκBα). Immunofluorescence microscopy showed that RGFP109 does not block the nuclear translocation of NF-κB/p65. However, co-immunoprecipitation assays revealed that RGFP109 induces the hyperacetylation of NF-κB/p65 and histones, and blocks interactions between NF-κB/p65 and its coactivators, p300 and p300/CBP-associated factor (PCAF). These results indicate that RGFP109-mediated post-translational nuclear acetylation may be involved in the regulation of NF-κB. Electrophoretic mobility shift assays revealed that RGFP109 reduces NF-κB/p65 binding to κB-DNA and decreased the transcriptional level of κB-mediated genes, suggesting that RGFP109-induced hyperacetylation leads to attenuated transcription of the κB gene. In addition, RGFP109 elevates the expression of inhibitor of growth 4 (ING4), which is typically downregulated in GBM cells. Importantly, we found that RGFP109 enhances ING4 recognition and binding to NF-κB/p65, which may be positively correlated with reduced interactions between NF-κB/p65 and p300/PCAF, thereby effecting transcription of the κB gene. Finally, we show that knockdown of ING4 with plasmids containing pcDNA3.1-ING4 shRNA abolished the effect of RGFP109. Therefore, ING4 may act as a corepressor and facilitate RGFP109-triggered suppression of the NF-κB pathway. Taken together, our data show that RGFP109, an HDAC inhibitor, in combination with TMZ may be a therapeutic candidate for patients with temozolomide-resistant GBM.

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References

  1. Stewart LA (2002) Chemotherapy in adult high-grade glioma: a systematic review and meta-analysis of individual patient data from 12 randomised trials. The Lancet 359:1011–1018

    Article  CAS  Google Scholar 

  2. Kokkinakis DM, Bocangel DB, Schold SC, Moschel RC, Pegg AE (2001) Thresholds of O6-alkylguanine-DNA alkyltransferase which confer significant resistance of human glial tumor xenografts to treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea or temozolomide. Clin Cancer Res 7:421–428

    CAS  PubMed  Google Scholar 

  3. Bredel M, Bredel C, Juric D, Duran GE, Yu RX, Harsh GR, Vogel H, Recht LD, Scheck AC, Sikic BI (2006) Tumor necrosis factor-alpha-induced protein 3 as a putative regulator of nuclear factor-κB-mediated resistance to O6-alkylating agents in human glioblastomas. J Clin Oncol 24:274–287

    Article  CAS  PubMed  Google Scholar 

  4. Atkinson GP, Nozell SE, Benveniste ET (2010) NF-κB and STAT3 signaling in glioma: targets for future therapies. Expert Rev Neurother 10:575–586

    Article  CAS  PubMed  Google Scholar 

  5. Nozell S, Laver T, Moseley D, Nowoslawski L, De Vos M, Atkinson GP, Harrison K, Nabors LB, Benveniste EN (2008) The ING4 tumor suppressor attenuates NF-κB activity at the promoters of target genes. Mol Cell Biol 28:6632–6645

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Kapoor GS, Zhan Y, Johnson GR, O’Rourke DM (2004) Distinct domains in the SHP-2 phosphatase differentially regulate epidermal growth factor receptor/NF-κB activation through Gab1 in glioblastoma cells. Mol Cell Biol 24:823–836

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Greene WC, Chen LF (2004) Regulation of NF-κB action by reversible acetylation. Novartis Found Symp 259:208–217 (discussion 218–225)

    Article  CAS  PubMed  Google Scholar 

  8. Vermeulen L, De Wilde G, Van Damme P, Berghe WV, Haegeman G (2003) Transcriptional activation of the NF-κB p65 subunit by mitogen- and stress-activated protein kinase-1 (MSK1). EMBO J 22:1313–1324

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Perkins ND, Gilmore TD (2006) Good cop, bad cop: the different faces of NF-κB. Cell Death Differ 13:759–772

    Article  CAS  PubMed  Google Scholar 

  10. Sheppard KA, Rose DW, Haque ZK, Kurokawa R, McInerney E, Westin S, Thanos D, Rosenfeld MG, Glass CK, Collins T (1999) Transcriptional activation by NF-κB requires multiple coactivators. Mol Cell Biol 19:6367–6378

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Zhong H, Voll RE, Ghosh S (1998) Phosphorylation of NF-κB p65 by PKA stimulates transcriptional activity by promoting a novel bivalent interaction with the coactivator CBP/p300. Mol Cell 1:661–671

    Article  CAS  PubMed  Google Scholar 

  12. Chen L, Fischle W, Verdin E, Greene WC (2001) Duration of nuclear NF-κB action regulated by reversible acetylation. Science 293:1653–1657

    Article  CAS  Google Scholar 

  13. Kiernan R, Bres V, Ng RW, Coudart MP, El Messaoudi S, Sardet C, Jin DY, Emiliani S, Benkirane M (2003) Post-activation turn-off of NF-κB-dependent transcription is regulated by acetylation of p65. J Biol Chem 278:2758–2766

    Article  CAS  PubMed  Google Scholar 

  14. Saito A, Yamashita T, Mariko Y, Nosaka Y, Tsuchiya K, Ando T, Suzuki T, Tsuruo T, Nakanishi O (1999) A synthetic inhibitor of histone deacetylase, MS-27-275, with marked in vivo antitumor activity against human tumors. Proc Natl Acad Sci USA 96:4592–4597

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Malvaez M, McQuown SC, Rogge GA, Astarabadi M, Jacques V, Carreiro S, Rusche JR, Wood MA (2013) HDAC3-selective inhibitor enhances extinction of cocaine-seeking behavior in a persistent manner. Proc Natl Acad Sci USA 110:2647–2652

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Johnston TH, Huot P, Damude S, Fox SH, Jones SW, Rusche JR, Brotchie JM (2013) RGFP109, a histone deacetylase inhibitor attenuates l-DOPA-induced dyskinesia in the MPTP-lesioned marmoset: a proof-of-concept study. Parkinsonism Relat Disord 19:260–264

    Article  PubMed  Google Scholar 

  17. Garkavtsev I, Kozin SV, Chernova O, Xu L, Winkler F, Brown E, Barnett GH, Jain RK (2004) The candidate tumour suppressor protein ING4 regulates brain tumour growth and angiogenesis. Nature 428:328–332

    Article  CAS  PubMed  Google Scholar 

  18. Liu E, Wu J, Cao W, Zhang J, Liu W, Jiang X, Zhang X (2007) Curcumin induces G2/M cell cycle arrest in a p53-dependent manner and upregulates ING4 expression in human glioma. J Neurooncol 85:263–270

    Article  CAS  PubMed  Google Scholar 

  19. Perkins ND (2006) Post-translational modifications regulating the activity and function of the nuclear factor κB pathway. Oncogene 25:6717–6730

    Article  CAS  PubMed  Google Scholar 

  20. Wolffe AP (1996) Histone deacetylase: a regulator of transcription. Science 272:371–372

    Article  CAS  PubMed  Google Scholar 

  21. Vega RB, Matsuda K, Oh J, Barbosa AC, Yang X, Meadows E, McAnally J, Pomajzl C, Shelton JM, Richardson JA, Karsenty G, Olson EN (2004) Histone deacetylase 4 controls chondrocyte hypertrophy during skeletogenesis. Cell 119:555–566

    Article  CAS  PubMed  Google Scholar 

  22. Sebastian C, Zwaans BM, Silberman DM, Gymrek M, Goren A, Zhong L, Ram O, Truelove J, Guimaraes AR, Toiber D, Cosentino C, Greenson JK, MacDonald AI, McGlynn L, Maxwell F, Edwards J, Giacosa S, Guccione E, Weissleder R, Bernstein BE, Regev A, Shiels PG, Lombard DB, Mostoslavsky R (2012) The histone deacetylase SIRT6 is a tumor suppressor that controls cancer metabolism. Cell 151:1185–1199

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Hu E, Dul E, Sung CM, Chen Z, Kirkpatrick R, Zhang GF, Johanson K, Liu R, Lago A, Hofmann G, Macarron R, de los Frailes M, Perez P, Krawiec J, Winkler J, Jaye M (2003) Identification of novel isoform-selective inhibitors within class I histone deacetylases. J Pharmacol Exp Therap 307:720–728

    Article  CAS  Google Scholar 

  24. Werner SL, Barken D, Hoffmann A (2005) Stimulus specificity of gene expression programs determined by temporal control of IKK activity. Science 309:1857–1861

    Article  CAS  PubMed  Google Scholar 

  25. Furia B, Deng L, Wu K, Baylor S, Kehn K, Li H, Donnelly R, Coleman T, Kashanchi F (2002) Enhancement of nuclear factor-κB acetylation by coactivator p300 and HIV-1 Tat proteins. J Biol Chem 277:4973–4980

    Article  CAS  PubMed  Google Scholar 

  26. Quivy V, Van Lint C (2004) Regulation at multiple levels of NF-κB-mediated transactivation by protein acetylation. Biochem Pharmacol 68:1221–1229

    Article  CAS  PubMed  Google Scholar 

  27. Hoberg JE, Yeung F, Mayo MW (2004) SMRT derepression by the IκB kinase alpha: a prerequisite to NF-κB transcription and survival. Mol Cell 16:245–255

    Article  CAS  PubMed  Google Scholar 

  28. Roth SY, Denu JM, Allis CD (2001) Histone acetyltransferases. Annu Rev Biochem 70:81–120

    Article  CAS  PubMed  Google Scholar 

  29. Vanden Berghe W, De Bosscher K, Boone E, Plaisance S, Haegeman G (1999) The nuclear factor-κB engages CBP/p300 and histone acetyltransferase activity for transcriptional activation of the interleukin-6 gene promoter. J Biol Chem 274:32091–32098

    Article  CAS  PubMed  Google Scholar 

  30. Kaina B, Christmann M, Naumann S, Roos WP (2007) MGMT: key node in the battle against genotoxicity, carcinogenicity and apoptosis induced by alkylating agents. DNA Rep 6:1079–1099

    Article  CAS  Google Scholar 

  31. Suka N, Carmen AA, Rundlett SE, Grunstein M (1998) The regulation of gene activity by histones and the histone deacetylase RPD3. Cold Spring Harb Symp Quant Biol 63:391–399

    Article  CAS  PubMed  Google Scholar 

  32. Camphausen K, Burgan W, Cerra M, Oswald KA, Trepel JB, Lee MJ, Tofilon PJ (2004) Enhanced radiation-induced cell killing and prolongation of \(\gamma\)H2AX foci expression by the histone deacetylase inhibitor MS-275. Cancer Res 64:316–321

    Article  CAS  PubMed  Google Scholar 

  33. Eyupoglu IY, Hahnen E, Trankle C, Savaskan NE, Siebzehnrubl FA, Buslei R, Lemke D, Wick W, Fahlbusch R, Blumcke I (2006) Experimental therapy of malignant gliomas using the inhibitor of histone deacetylase MS-275. Mol Cancer Ther 5:1248–1255

    Article  PubMed  Google Scholar 

  34. Palacios A, Garcia P, Padro D, Lopez-Hernandez E, Martin I, Blanco FJ (2006) Solution structure and NMR characterization of the binding to methylated histone tails of the plant homeodomain finger of the tumour suppressor ING4. FEBS Lett 580:6903–6908

    Article  CAS  PubMed  Google Scholar 

  35. Pena PV, Davrazou F, Shi X, Walter KL, Verkhusha VV, Gozani O, Zhao R, Kutateladze TG (2006) Molecular mechanism of histone H3K4me3 recognition by plant homeodomain of ING2. Nature 442:100–103

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Gong W, Suzuki K, Russell M, Riabowol K (2005) Function of the ING family of PHD proteins in cancer. Int J Biochem Cell Biol 37:1054–1065

    Article  CAS  PubMed  Google Scholar 

  37. Campos EI, Chin MY, Kuo WH, Li G (2004) Biological functions of the ING family tumor suppressors. Cell Mol Life Sci 61:2597–2613

    Article  CAS  PubMed  Google Scholar 

  38. Shi X, Hong T, Walter KL, Ewalt M, Michishita E, Hung T, Carney D, Pena P, Lan F, Kaadige MR, Lacoste N, Cayrou C, Davrazou F, Saha A, Cairns BR, Ayer DE, Kutateladze TG, Shi Y, Cote J, Chua KF, Gozani O (2006) ING2 PHD domain links histone H3 lysine 4 methylation to active gene repression. Nature 442:96–99

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Doyon Y, Cayrou C, Ullah M, Landry AJ, Cote V, Selleck W, Lane WS, Tan S, Yang XJ, Cote J (2006) ING tumor suppressor proteins are critical regulators of chromatin acetylation required for genome expression and perpetuation. Mol Cell 21:51–64

    Article  CAS  PubMed  Google Scholar 

  40. Shiseki M, Nagashima M, Pedeux RM, Kitahama-Shiseki M, Miura K, Okamura S, Onogi H, Higashimoto Y, Appella E, Yokota J, Harris CC (2003) p29ING4 and p28ING5 bind to p53 and p300, and enhance p53 activity. Cancer Res 63:2373–2378

    CAS  PubMed  Google Scholar 

  41. Zhang X, Wang KS, Wang ZQ, Xu LS, Wang QW, Chen F, Wei DZ, Han ZG (2005) Nuclear localization signal of ING4 plays a key role in its binding to p53. Biochem Biophys Res Commun 331:1032–1038

    Article  CAS  PubMed  Google Scholar 

  42. Gunduz M, Nagatsuka H, Demircan K, Gunduz E, Cengiz B, Ouchida M, Tsujigiwa H, Yamachika E, Fukushima K, Beder L, Hirohata S, Ninomiya Y, Nishizaki K, Shimizu K, Nagai N (2005) Frequent deletion and down-regulation of ING4, a candidate tumor suppressor gene at 12p13, in head and neck squamous cell carcinomas. Gene 356:109–117

    Article  CAS  PubMed  Google Scholar 

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    Authors and Affiliations

    1. Brain Center, Department of Neurosurgery, Shenzhen Key Laboratory of Neurosurgery, Shenzhen Second People’s Hospital, The Clinical College of Anhui Medical University, Shenzhen University 1st Affiliated Hospital, 3002# Sungang Road, Futian district, Shenzhen, 518035, China

      Zong-yang Li, Qing-zhong Li, Lei Chen, Bao-dong Chen, Bo Wang, Xie-jun Zhang & Wei-ping Li

    2. Shantou University Medical College, 22# Xinling Road, Shantou, Guangdong, China

      Qing-zhong Li & Wei-ping Li

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    1. Zong-yang Li
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    Zong-yang Li and Qing-zhong Li have contributed equally to this work.

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    Li, Zy., Li, Qz., Chen, L. et al. Histone Deacetylase Inhibitor RGFP109 Overcomes Temozolomide Resistance by Blocking NF-κB-Dependent Transcription in Glioblastoma Cell Lines. Neurochem Res 41, 3192–3205 (2016). https://doi.org/10.1007/s11064-016-2043-5

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