Dual Targeting of G9a and DNA Methyltransferase-1 for the Treatment of Experimental Cholangiocarcinoma.

Animals Bile Duct Neoplasms / drug therapy CCAAT-Enhancer-Binding Proteins / metabolism Cell Line, Tumor Cell Proliferation / drug effects Cholangiocarcinoma / drug therapy DNA (Cytosine-5-)-Methyltransferase 1 / antagonists & inhibitors DNA Methylation / drug effects Enzyme Inhibitors / pharmacology Epigenesis, Genetic / drug effects Gene Expression Regulation, Neoplastic / drug effects Histocompatibility Antigens / metabolism Histone Code / drug effects Histone-Lysine N-Methyltransferase / antagonists & inhibitors Humans Mice Treatment Outcome Ubiquitin-Protein Ligases / metabolism Xenograft Model Antitumor Assays / methods

Journal

Hepatology (Baltimore, Md.)

ISSN: 1527-3350

Titre abrégé: Hepatology

Pays: United States

ID NLM: 8302946

Informations de publication

Date de publication:
06 2021

Historique:

revised: 06 10 2020

received: 05 02 2020

accepted: 14 10 2020

pubmed: 23 11 2020

medline: 8 1 2022

entrez: 22 11 2020

Statut: ppublish

Résumé

Cholangiocarcinoma (CCA) is a devastating disease often detected at advanced stages when surgery cannot be performed. Conventional and targeted systemic therapies perform poorly, and therefore effective drugs are urgently needed. Different epigenetic modifications occur in CCA and contribute to malignancy. Targeting epigenetic mechanisms may thus open therapeutic opportunities. However, modifications such as DNA and histone methylation often coexist and cooperate in carcinogenesis. We tested the therapeutic efficacy and mechanism of action of a class of dual G9a histone-methyltransferase and DNA-methyltransferase 1 (DNMT1) inhibitors. Expression of G9a, DNMT1, and their molecular adaptor, ubiquitin-like with PHD and RING finger domains-1 (UHRF1), was determined in human CCA. We evaluated the effect of individual and combined pharmacological inhibition of G9a and DNMT1 on CCA cell growth. Our lead G9a/DNMT1 inhibitor, CM272, was tested in human CCA cells, patient-derived tumoroids and xenograft, and a mouse model of cholangiocarcinogenesis with hepatocellular deletion of c-Jun-N-terminal-kinase (Jnk)-1/2 and diethyl-nitrosamine (DEN) plus CCl Dual targeting of G9a and DNMT1 with epigenetic small molecule inhibitors such as CM272 is a potential strategy to treat CCA and/or enhance the efficacy of other systemic therapies.

Sections du résumé

BACKGROUND AND AIMS

APPROACH AND RESULTS

Expression of G9a, DNMT1, and their molecular adaptor, ubiquitin-like with PHD and RING finger domains-1 (UHRF1), was determined in human CCA. We evaluated the effect of individual and combined pharmacological inhibition of G9a and DNMT1 on CCA cell growth. Our lead G9a/DNMT1 inhibitor, CM272, was tested in human CCA cells, patient-derived tumoroids and xenograft, and a mouse model of cholangiocarcinogenesis with hepatocellular deletion of c-Jun-N-terminal-kinase (Jnk)-1/2 and diethyl-nitrosamine (DEN) plus CCl

CONCLUSIONS

Dual targeting of G9a and DNMT1 with epigenetic small molecule inhibitors such as CM272 is a potential strategy to treat CCA and/or enhance the efficacy of other systemic therapies.

Identifiants

DOI: 10.1002/hep.31642 PMID: 33222246

pubmed: 33222246

doi: 10.1002/hep.31642

doi:

Substances chimiques

CCAAT-Enhancer-Binding Proteins 0

Enzyme Inhibitors 0

Histocompatibility Antigens 0

DNA (Cytosine-5-)-Methyltransferase 1 EC 2.1.1.37

DNMT1 protein, human EC 2.1.1.37

EHMT2 protein, human EC 2.1.1.43

Histone-Lysine N-Methyltransferase EC 2.1.1.43

UHRF1 protein, human EC 2.3.2.27

Ubiquitin-Protein Ligases EC 2.3.2.27

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

Pagination

2380-2396

Informations de copyright

Références

Banales JM, Cardinale V, Carpino G, Marzioni M, Andersen JB, Invernizzi P, et al. Cholangiocarcinoma: current knowledge and future perspectives consensus statement from the European Network for the Study of Cholangiocarcinoma (ENS-CCA). Nat Rev Gastroenterol Hepatol 2016;13:261-280.

Rizvi S, Khan SA, Hallemeier CL, Kelley RK, Gores GJ. Cholangiocarcinoma-evolving concepts and therapeutic strategies. Nat Rev Clin Oncol 2018;15:95-111.

Nakamura H, Arai Y, Totoki Y, Shirota T, Elzawahry A, Kato M, et al. Genomic spectra of biliary tract cancer. Nat Genet 2015;47:1003-1010.

Marin JJG, Lozano E, Herraez E, Asensio M, Di Giacomo S, Romero MR, et al. Chemoresistance and chemosensitization in cholangiocarcinoma. Biochim Biophys Acta Mol Basis Dis 2018;1864:1444-1453.

Farshidfar F, Zheng S, Gingras MC, Newton Y, Shih J, Robertson AG, et al. Integrative genomic analysis of cholangiocarcinoma identifies distinct IDH-mutant molecular profiles. Cell Rep 2017;19:2878-2880.

O’Rourke CJ, Lafuente-Barquero J, Andersen JB. Epigenome remodeling in cholangiocarcinoma. Trends Cancer 2019;5:335-350.

Goeppert B, Konermann C, Schmidt CR, Bogatyrova O, Geiselhart L, Ernst C, et al. Global alterations of DNA methylation in cholangiocarcinoma target the Wnt signaling pathway. Hepatology 2014;59:544-554.

Merino-Azpitarte M, Lozano E, Perugorria MJ, Esparza-Baquer A, Erice O, Santos-Laso Á, et al. SOX17 regulates cholangiocyte differentiation and acts as a tumor suppressor in cholangiocarcinoma. J Hepatol 2017;67:72-83.

Nakamura K, Nakabayashi K, Htet Aung K, Aizawa K, Hori N, Yamauchi J, et al. DNA methyltransferase inhibitor zebularine induces human cholangiocarcinoma cell death through alteration of DNA methylation status. PLoS One 2015;10:e0120545.

Tariq NUA, McNamara MG, Valle JW. Biliary tract cancers: current knowledge, clinical candidates and future challenges. Cancer Manag Res 2019;11:2623-2642.

Nakagawa S, Okabe H, Sakamoto Y, Hayashi H, Hashimoto D, Yokoyama N, et al. Enhancer of zeste homolog 2 (EZH2) promotes progression of cholangiocarcinoma cells by regulating cell cycle and apoptosis. Ann Surg Oncol 2013;20(Suppl 3):S667-S675.

Du J, Johnson LM, Jacobsen SE, Patel DJ. DNA methylation pathways and their crosstalk with histone methylation. Nat Rev Mol Cell Biol 2015;16:519-532.

Casciello F, Windloch K, Gannon F, Lee JS. Functional role of G9a histone methyltransferase in cancer. Front Immunol 2015;6:487.

Wozniak RJ, Klimecki WT, Lau SS, Feinstein Y, Futscher BW. 5-Aza-2’-deoxycytidine-mediated reductions in G9A histone methyltransferase and histone H3 K9 di-methylation levels are linked to tumor suppressor gene reactivation. Oncogene 2007;26:77-90.

Ferry L, Fournier A, Tsusaka T, Adelmant G, Shimazu T, Matano S, et al. Methylation of DNA ligase 1 by G9a/GLP recruits UHRF1 to replicating DNA and regulates DNA methylation. Mol Cell 2017;67:550-565.e5.

Xue B, Zhao J, Feng P, Xing J, Wu H, Li Y. Epigenetic mechanism and target therapy of uhrf1 protein complex in malignancies. Onco Targets Ther 2019;12:549-559.

San José-Enériz E, Agirre X, Rabal O, Vilas-Zornoza A, Sanchez-Arias JA, Miranda E, et al. Discovery of first-in-class reversible dual small molecule inhibitors against G9a and DNMTs in hematological malignancies. Nat Commun 2017;8:15424.

Bárcena-Varela M, Caruso S, Llerena S, Álvarez-Sola G, Uriarte I, Latasa MU, et al. Dual targeting of histone methyltransferase G9a and DNA-methyltransferase 1 for the treatment of experimental hepatocellular carcinoma. Hepatology 2019;69:587-603.

Cubero FJ, Mohamed MR, Woitok MM, Zhao G, Hatting M, Nevzorova YA, et al. Loss of c-Jun N-terminal kinase 1 and 2 function in liver epithelial cells triggers biliary hyperproliferation resembling cholangiocarcinoma. Hepatol Commun 2020;4:834-851.

Manieri E, Folgueira C, Rodríguez ME, Leiva-Vega L, Esteban-Lafuente L, Chen C, et al. JNK-mediated disruption of bile acid homeostasis promotes intrahepatic cholangiocarcinoma. Proc Natl Acad Sci 2020;117:16492-16499.

Mayr C, Helm K, Jakab M, Ritter M, Shrestha R, Makaju R, et al. The histone methyltransferase G9a: a new therapeutic target in biliary tract cancer. Hum Pathol 2018;72:117-126.

Zhu M, Wei C, Lin J, Dong S, Gao D, Chen J, et al. UHRF1 is regulated by miR-124-3p and promotes cell proliferation in intrahepatic cholangiocarcinoma. J Cell Physiol 2019;234:19875-19885.

Zach S, Birgin E, Rückert F. Primary cholangiocellular carcinoma cell lines. J Stem Cell Res Transplant 2015;2:1013.

Broutier L, Mastrogiovanni G, Verstegen MMA, Francies HE, Gavarró LM, Bradshaw CR, et al. Human primary liver cancer-derived organoid cultures for disease modeling and drug screening. Nat Med 2017;23:1424-1435.

Wong N, Li L, Tsang K, Lai PBS, To KF, Johnson PJ. Frequent loss of chromosome 3p and hypermethylation of RASSF1A in cholangiocarcinoma. J Hepatol 2002;37:633-639.

Malpeli G, Innamorati G, Decimo I, Bencivenga M, Nwabo Kamdje AH, Perris R, et al. Methylation dynamics of RASSF1A and its impact on cancer. Cancers (Basel) 2019;11:959.

Andersen JB, Spee B, Blechacz BR, Avital I, Komuta M, Barbour A, et al. Genomic and genetic characterization of cholangiocarcinoma identifies therapeutic targets for tyrosine kinase inhibitors. Gastroenterology 2012;142:1021-1031.e15.

Wood ER, Truesdale AT, McDonald OB, Yuan D, Hassell A, Dickerson SH, et al. A unique structure for epidermal growth factor receptor bound to GW572016 (Lapatinib): relationships among protein conformation, inhibitor off-rate, and receptor activity in tumor cells. Cancer Res 2004;64:6652-6659.

Mu X, Pradere JP, Affò S, Dapito DH, Friedman R, Lefkovitch JH, et al. Epithelial transforming growth factor-β signaling does not contribute to liver fibrosis but protects mice from Cholangiocarcinoma. Gastroenterology 2016;150:720-733.

Pavlova NN, Thompson CB. The emerging hallmarks of cancer metabolism. Cell Metab 2016;23:27-47.

Satriano L, Lewinska M, Rodrigues PM, Banales JM, Andersen JB. Metabolic rearrangements in primary liver cancers: cause and consequences. Nat Rev Gastroenterol Hepatol 2019;16:748-766.

Reina-Campos M, Diaz-Meco MT, Moscat J. The complexity of the serine glycine one-carbon pathway in cancer. J Cell Biol 2020;219:e201907022.

Huang YC, Chen M, Shyr YM, Su CH, Chen CK, Li AFY, et al. Glycine N-methyltransferase is a favorable prognostic marker for human cholangiocarcinoma. J Gastroenterol Hepatol 2008;23:1384-1389.

Yang H, Liu T, Wang J, Li TWH, Fan W, Peng H, et al. Deregulated methionine adenosyltransferase α1, c-Myc, and Maf proteins together promote cholangiocarcinoma growth in mice and humans. Hepatology 2016;64:439-455.

Zhao W, Yang S, Chen J, Zhao J, Dong J. Forced overexpression of FBP1 inhibits proliferation and metastasis in cholangiocarcinoma cells via Wnt/β-catenin pathway. Life Sci 2018;210:224-234.

Liu D, Wong CC, Fu L, Chen H, Zhao L, Li C, et al. Squalene epoxidase drives NAFLD-induced hepatocellular carcinoma and is a pharmaceutical target. Sci Transl Med 2018;10:eaap9840.

Suzuki H, Komuta M, Bolog A, Yokobori T, Wada S, Araki K, et al. Relationship between 18-F-fluoro-deoxy-D-glucose uptake and expression of glucose transporter 1 and pyruvate kinase M2 in intrahepatic cholangiocarcinoma. Dig Liver Dis 2015;47:590-596.

Ponnusamy L, Mahalingaiah PKS, Singh KP. Epigenetic reprogramming and potential application of epigenetic-modifying drugs in acquired chemotherapeutic resistance. Adv Clin Chem 2020;94:219-259.

Ma W, Han C, Zhang J, Song K, Chen W, Kwon H, et al. The histone methyltransferase G9a promotes cholangiocarcinogenesis through regulation of the Hippo pathway kinase LATS2 and YAP signaling pathway. Hepatology 2020;72:1283-1297.

Tan J, Yang X, Zhuang L, Jiang X, Chen W, Puay LL, et al. Pharmacologic disruption of polycomb-repressive complex 2-mediated gene repression selectively induces apoptosis in cancer cells. Genes Dev 2007;21:1050-1063.

Purcell DJ, Khalid O, Ou CY, Little GH, Frenkel B, Baniwal SK, et al. Recruitment of coregulator G9a by Runx2 for selective enhancement or suppression of transcription. J Cell Biochem 2012;113:2406-2414.

Fang F, Munck J, Tang J, Taverna P, Wang Y, Miller DFB, et al. The novel, small-molecule DNA methylation inhibitor SGI-110 as an ovarian cancer chemosensitizer. Clin Cancer Res 2014;20:6504-6516.

Lozano E, Asensio M, Perez-Silva L, Banales JM, Briz O, Marin JJG. MRP3-mediated chemoresistance in cholangiocarcinoma: target for chemosensitization through restoring SOX17 expression. Hepatology 2020;72:949-964.

Lozano E, Macias RIR, Monte MJ, Asensio M, Carmen S, Sanchez-Vicente L, et al. Causes of hOCT 1-dependent cholangiocarcinoma resistance to sorafenib and sensitization by tumor-selective gene therapy. Hepatology 2019;70:1246-1261.

Cirmena G, Franceschelli P, Isnaldi E, Ferrando L, De Mariano M, Ballestrero A, et al. Squalene epoxidase as a promising metabolic target in cancer treatment. Cancer Lett 2018;425:13-20.

Xu L, Wang L, Zhou L, Dorfman RG, Pan Y, Tang D, et al. The SIRT2/cMYC pathway inhibits peroxidation-related apoptosis in cholangiocarcinoma through metabolic reprogramming. Neoplasia 2019;21:429-441.

Dong C, Yuan T, Wu Y, Wang Y, Fan T, Miriyala S, et al. Loss of FBP1 by Snail-mediated repression provides metabolic advantages in basal-like breast cancer. Cancer Cell 2013;23:316-331.

Tu WB, Shiah YJ, Lourenco C, Mullen PJ, Dingar D, Redel C, et al. MYC interacts with the G9a histone methyltransferase to drive transcriptional repression and tumorigenesis. Cancer Cell 2018;34:579-595.e8.

Wang B, Fan P, Zhao J, Wu H, Jin X, Wu H. FBP1 loss contributes to BET inhibitors resistance by undermining c-Myc expression in pancreatic ductal adenocarcinoma. J Exp Clin Cancer Res 2018;37:224.

De Thé H. Differentiation therapy revisited. Nat Rev Cancer 2018;18:117-127.