Abstract
New therapeutics directed against established and novel molecular targets are urgently needed to intervene against cancer. Recently, it was reported that several members of the sirtuin family (SIRT1–7), the mammalian orthologs of the silent information regulator 2 (Sir2) protein in Saccharomyces cerevisiae, play important roles in carcinogenesis. Although SIRT2 has been attributed both tumor-promoting and tumor-suppressing activities in different contexts, selective SIRT2 inhibition with a small molecule mechanism-based inhibitor known as Thiomyristoyl lysine (TM) repressed the growth of breast cancer cell lines. In light of the anticancer effect of SIRT2 inhibition in cell culture, it was critical to assess the efficacy of TM as a potential anticancer therapy in vivo. This was accomplished by testing the SIRT2 inhibitor in genetically engineered and xenotransplantation mouse models of breast cancer, using the procedures detailed in this chapter.
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References
National Cancer Institute (2018) Cancer statistics. https://www.cancer.gov/about-cancer/understanding/statistics. Accessed 27 Mar 2018
Arif M, Senapati P, Shandilya J, Kundu TK (2010) Protein lysine acetylation in cellular function and its role in cancer manifestation. Biochim Biophys Acta 1799(10–12):702–716. https://doi.org/10.1016/j.bbagrm.2010.10.002
Krueger KE, Srivastava S (2006) Posttranslational protein modifications: current implications for cancer detection, prevention, and therapeutics. Mol Cell Proteomics 5(10):1799–1810. https://doi.org/10.1074/mcp.R600009-MCP200
Chalkiadaki A, Guarente L (2015) The multifaceted functions of sirtuins in cancer. Nat Rev Cancer 15(10):608–624. https://doi.org/10.1038/nrc3985
Chang HC, Guarente L (2014) SIRT1 and other sirtuins in metabolism. Trends Endocrinol Metab 25(3):138–145. https://doi.org/10.1016/j.tem.2013.12.001
Haigis MC, Sinclair DA (2010) Mammalian sirtuins: biological insights and disease relevance. Annu Rev Pathol 5:253–295. https://doi.org/10.1146/annurev.pathol.4.110807.092250
Lavu S, Boss O, Elliott PJ, Lambert PD (2008) Sirtuins--novel therapeutic targets to treat age-associated diseases. Nat Rev Drug Discov 7(10):841–853. https://doi.org/10.1038/nrd2665
Blander G, Guarente L (2004) The Sir2 family of protein deacetylases. Annu Rev Biochem 73:417–435. https://doi.org/10.1146/annurev.biochem.73.011303.073651
Imai S, Guarente L (2010) Ten years of NAD-dependent SIR2 family deacetylases: implications for metabolic diseases. Trends Pharmacol Sci 31(5):212–220. https://doi.org/10.1016/j.tips.2010.02.003
Houtkooper RH, Pirinen E, Auwerx J (2012) Sirtuins as regulators of metabolism and healthspan. Nat Rev Mol Cell Biol 13(4):225–238. https://doi.org/10.1038/nrm3293
Imai S, Armstrong CM, Kaeberlein M, Guarente L (2000) Transcriptional silencing and longevity protein Sir2 is an NAD-dependent histone deacetylase. Nature 403(6771):795–800. https://doi.org/10.1038/35001622
de Oliveira RM, Sarkander J, Kazantsev AG, Outeiro TF (2012) SIRT2 as a therapeutic target for age-related disorders. Front Pharmacol 3:82. https://doi.org/10.3389/fphar.2012.00082
Park S-H, Zhu Y, Ozden O, Kim H-S, Jiang H, Deng C-X, Gius D, Vassilopoulos A (2012) SIRT2 is a tumor suppressor that connects aging, acetylome, cell cycle signaling, and carcinogenesis. Transl Cancer Res 1(1):15–21
Zhou W, Ni TK, Wronski A, Glass B, Skibinski A, Beck A, Kuperwasser C (2016) The SIRT2 deacetylase stabilizes slug to control malignancy of basal-like breast cancer. Cell Rep 17(5):1302–1317. https://doi.org/10.1016/j.celrep.2016.10.006
Serrano L, Martinez-Redondo P, Marazuela-Duque A, Vazquez BN, Dooley SJ, Voigt P, Beck DB, Kane-Goldsmith N, Tong Q, Rabanal RM, Fondevila D, Munoz P, Kruger M, Tischfield JA, Vaquero A (2013) The tumor suppressor SirT2 regulates cell cycle progression and genome stability by modulating the mitotic deposition of H4K20 methylation. Genes Dev 27(6):639–653. https://doi.org/10.1101/gad.211342.112
McGlynn LM, Zino S, MacDonald AI, Curle J, Reilly JE, Mohammed ZM, McMillan DC, Mallon E, Payne AP, Edwards J, Shiels PG (2014) SIRT2: tumour suppressor or tumour promoter in operable breast cancer? Eur J Cancer 50(2):290–301. https://doi.org/10.1016/j.ejca.2013.10.005
Jing H, Hu J, He B, Negron Abril YL, Stupinski J, Weiser K, Carbonaro M, Chiang YL, Southard T, Giannakakou P, Weiss RS, Lin H (2016) A SIRT2-selective inhibitor promotes c-Myc oncoprotein degradation and exhibits broad anticancer activity. Cancer Cell 29(3):297–310. https://doi.org/10.1016/j.ccell.2016.02.007
Chen J, Chan AW, To KF, Chen W, Zhang Z, Ren J, Song C, Cheung YS, Lai PB, Cheng SH, Ng MH, Huang A, Ko BC (2013) SIRT2 overexpression in hepatocellular carcinoma mediates epithelial to mesenchymal transition by protein kinase B/glycogen synthase kinase-3beta/beta-catenin signaling. Hepatology 57(6):2287–2298. https://doi.org/10.1002/hep.26278
Cheon MG, Kim W, Choi M, Kim JE (2015) AK-1, a specific SIRT2 inhibitor, induces cell cycle arrest by downregulating Snail in HCT116 human colon carcinoma cells. Cancer Lett 356(2 Pt B):637–645. https://doi.org/10.1016/j.canlet.2014.10.012
He B, Hu J, Zhang X, Lin H (2014) Thiomyristoyl peptides as cell-permeable Sirt6 inhibitors. Org Biomol Chem 12(38):7498–7502. https://doi.org/10.1039/c4ob00860j
Heltweg B, Gatbonton T, Schuler AD, Posakony J, Li H, Goehle S, Kollipara R, Depinho RA, Gu Y, Simon JA, Bedalov A (2006) Antitumor activity of a small-molecule inhibitor of human silent information regulator 2 enzymes. Cancer Res 66(8):4368–4377. https://doi.org/10.1158/0008-5472.Can-05-3617
Hoffmann G, Breitenbucher F, Schuler M, Ehrenhofer-Murray AE (2014) A novel sirtuin 2 (SIRT2) inhibitor with p53-dependent pro-apoptotic activity in non-small cell lung cancer. J Biol Chem 289(8):5208–5216. https://doi.org/10.1074/jbc.M113.487736
Kim WJ, Lee JW, Quan C, Youn HJ, Kim HM, Bae SC (2011) Nicotinamide inhibits growth of carcinogen induced mouse bladder tumor and human bladder tumor xenograft through up-regulation of RUNX3 and p300. J Urol 185(6):2366–2375. https://doi.org/10.1016/j.juro.2011.02.017
Mahajan SS, Scian M, Sripathy S, Posakony J, Lao U, Loe TK, Leko V, Thalhofer A, Schuler AD, Bedalov A, Simon JA (2014) Development of pyrazolone and isoxazol-5-one cambinol analogues as sirtuin inhibitors. J Med Chem 57(8):3283–3294. https://doi.org/10.1021/jm4018064
McCarthy AR, Sachweh MC, Higgins M, Campbell J, Drummond CJ, van Leeuwen IM, Pirrie L, Ladds MJ, Westwood NJ, Lain S (2013) Tenovin-D3, a novel small-molecule inhibitor of sirtuin SirT2, increases p21 (CDKN1A) expression in a p53-independent manner. Mol Cancer Ther 12(4):352–360. https://doi.org/10.1158/1535-7163.Mct-12-0900
Neugebauer RC, Uchiechowska U, Meier R, Hruby H, Valkov V, Verdin E, Sippl W, Jung M (2008) Structure-activity studies on splitomicin derivatives as sirtuin inhibitors and computational prediction of binding mode. J Med Chem 51(5):1203–1213. https://doi.org/10.1021/jm700972e
Rotili D, Tarantino D, Nebbioso A, Paolini C, Huidobro C, Lara E, Mellini P, Lenoci A, Pezzi R, Botta G, Lahtela-Kakkonen M, Poso A, Steinkuhler C, Gallinari P, De Maria R, Fraga M, Esteller M, Altucci L, Mai A (2012) Discovery of salermide-related sirtuin inhibitors: binding mode studies and antiproliferative effects in cancer cells including cancer stem cells. J Med Chem 55(24):10937–10947. https://doi.org/10.1021/jm3011614
Zhang Y, Au Q, Zhang M, Barber JR, Ng SC, Zhang B (2009) Identification of a small molecule SIRT2 inhibitor with selective tumor cytotoxicity. Biochem Biophys Res Commun 386(4):729–733. https://doi.org/10.1016/j.bbrc.2009.06.113
Du J, Zhou Y, Su X, Yu JJ, Khan S, Jiang H, Kim J, Woo J, Kim JH, Choi BH, He B, Chen W, Zhang S, Cerione RA, Auwerx J, Hao Q, Lin H (2011) Sirt5 is a NAD-dependent protein lysine demalonylase and desuccinylase. Science 334(6057):806–809. https://doi.org/10.1126/science.1207861
Feldman JL, Baeza J, Denu JM (2013) Activation of the protein deacetylase SIRT6 by long-chain fatty acids and widespread deacylation by mammalian sirtuins. J Biol Chem 288(43):31350–31356. https://doi.org/10.1074/jbc.C113.511261
Zhu AY, Zhou Y, Khan S, Deitsch KW, Hao Q, Lin H (2012) Plasmodium falciparum Sir2A preferentially hydrolyzes medium and long chain fatty acyl lysine. ACS Chem Biol 7(1):155–159. https://doi.org/10.1021/cb200230x
Bheda P, Jing H, Wolberger C, Lin H (2016) The substrate specificity of sirtuins. Annu Rev Biochem 85:405–429. https://doi.org/10.1146/annurev-biochem-060815-014537
Hawse WF, Hoff KG, Fatkins D, Daines A, Zubkova OV, Schramm VL, Zheng W, Wolberger C (2008) Structural insights into intermediate steps in the Sir2 deacetylation reaction. Structure 16(9):1368–1377. https://doi.org/10.1016/j.str.2008.05.015
Cen Y, Falco JN, Xu P, Youn DY, Sauve AA (2011) Mechanism-based affinity capture of sirtuins. Org Biomol Chem 9(4):987–993. https://doi.org/10.1039/c0ob00774a
Guy CT, Cardiff RD, Muller WJ (1992) Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease. Mol Cell Biol 12(3):954–961
Kim WY, Sharpless NE (2012) Drug efficacy testing in mice. Curr Top Microbiol Immunol 355:19–38. https://doi.org/10.1007/82_2011_160
Institute for Laboratory Animal Research (2011) Guide for the care and use of laboratory animals, 8th edn. National Academies Press, Washington, DC
Faustino-Rocha A, Oliveira PA, Pinho-Oliveira J, Teixeira-Guedes C, Soares-Maia R, da Costa RG, Colaco B, Pires MJ, Colaco J, Ferreira R, Ginja M (2013) Estimation of rat mammary tumor volume using caliper and ultrasonography measurements. Lab Anim 42(6):217–224. https://doi.org/10.1038/laban.254
Earnest E, Ajaghaku D (2014) Guidelines on dosage calculation and stock solution preparation in experimental animals’ studies. J Nat Sci Res 4(18):100–106
Hollingshead MG (2008) Antitumor efficacy testing in rodents. J Natl Cancer Inst 100(21):1500–1510. https://doi.org/10.1093/jnci/djn351
Machholz E, Mulder G, Ruiz C, Corning BF, Pritchett-Corning KR (2012) Manual restraint and common compound administration routes in mice and rats. J Vis Exp (67):e2771. https://doi.org/10.3791/2771
Acknowledgments
This work was supported in part by an intercampus seed grant from Cornell University and NIH R01 grant CA163255. Y.L.N.A. and I.F. were supported by NIH grants T32 GM008500 and T32 GM007273, respectively. The authors thank Dr. Elizabeth Moore for comments on the manuscript, and Dr. Hening Lin and members of his laboratory for helpful discussions and providing TM compound.
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Negrón Abril, Y.L., Fernández, I., Weiss, R.S. (2019). Assessment of SIRT2 Inhibitors in Mouse Models of Cancer. In: Brosh, Jr., R. (eds) Protein Acetylation. Methods in Molecular Biology, vol 1983. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9434-2_9
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