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The Robust Tumoricidal Effects of Combined BET/HDAC Inhibition in Cutaneous T-Cell Lymphoma Can Be Reproduced by ΔNp73 Depletion

      Combined BET inhibitor/histone deacetylase inhibitor treatment induces marked apoptosis of cutaneous T-cell lymphoma (CTCL) with minimal normal T-cell toxicity. At 96 hours when apoptosis was extensive, a majority of CTCL lines showed ≥2-fold suppression of T-cell survival factors (e.g., AKT1, BCL2 antiapoptotic factors, BIRC5, CD40, CD70, GADD45A, PRKCA, TNFRSF1B, ΔNp73) and ≥2-fold upregulation of proapoptotic factors and tumor suppressors (e.g., ATM, BAK, BIM, multiple caspases, FHIT, HIC1, MGMT, NOD1) (P < 0.05). The largest alterations were in TP73 isoform expression, resulting in increased TAp73/ΔNp73 ratios in CTCL lines and leukemic Sézary cells. Targeted ΔNp73 inhibition by small interfering RNA knockdown resulted in robust CTCL apoptosis comparable with that induced by BET inhibitor/histone deacetylase inhibitor with minimal normal T-cell toxicity. Chromatin immunoprecipitation analysis showed that BET inhibitor/histone deacetylase inhibitor treatment reduced RNA polymerase II binding to ΔNp73, MYC, and AKT1 while increasing its binding to TAp73. CTCL skin lesions expressed both TAp73 and ΔNp73 isoforms in situ. In aggregate, these findings implicate TAp73/ΔNp73 balance as a major factor governing CTCL survival, show that the expression of p73 isoforms can be altered by molecular biological and pharmaceutical means, show that p73 isoforms are expressed across the entire CTCL clinical spectrum, and identify the p73 pathway as a potential target for therapeutics.

      Abbreviations:

      BETi (BET inhibitor), CTCL (cutaneous T-cell lymphoma), ERK (extracellular signal‒regulated kinase), HDACi (histone deacetylase inhibitor), TA (transactivating)
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      References

        • Acharya P.
        • Sampathi S.
        • Zhao Y.
        • Wang J.
        • Liu Q.
        • Stengel K.
        • et al.
        Histone deacetylase inhibitors mimic BET inhibitors and cause RNA polymerase pausing at MYC, KIT, and other oncogenes in t(8;21) AML.
        Blood. 2017; 130: 2470
        • Bailey S.G.
        • Cragg M.S.
        • Townsend P.A.
        Family friction as ΔNp73 antagonises p73 and p53.
        Int J Biochem Cell Biol. 2011; 43: 482-486
        • Bisso A.
        • Collavin L.
        • Del Sal G.
        p73 as a pharmaceutical target for cancer therapy.
        Curr Pharm Des. 2011; 17: 578-590
        • Braun F.K.
        • Al-Yacoub N.
        • Plötz M.
        • Möbs M.
        • Sterry W.
        • Eberle J.
        Nonsteroidal anti-inflammatory drugs induce apoptosis in cutaneous T-cell lymphoma cells and enhance their sensitivity for TNF-related apoptosis-inducing ligand.
        J Invest Dermatol. 2012; 132: 429-439
        • Brunner P.M.
        • Jonak C.
        • Knobler R.
        Recent advances in understanding and managing cutaneous T-cell lymphomas.
        F1000Res. 2020; 9 (F1000 Faculty Rev-331)
        • Buhlmann S.
        • Pützer B.M.
        DNp73 a matter of cancer: mechanisms and clinical implications.
        Biochim Biophys Acta. 2008; 1785: 207-216
        • Chakraborty A.R.
        • Robey R.W.
        • Luchenko V.L.
        • Zhan Z.
        • Piekarz R.L.
        • Gillet J.P.
        • et al.
        MAPK pathway activation leads to Bim loss and histone deacetylase inhibitor resistance: rationale to combine Romidepsin with an MEK inhibitor.
        Blood. 2013; 121: 4115-4125
        • Di C.
        • Yang L.
        • Zhang H.
        • Ma X.
        • Zhang X.
        • Sun C.
        • et al.
        Mechanisms, function and clinical applications of DNp73.
        Cell Cycle. 2013; 12: 1861-1867
        • Domínguez G.
        • García J.M.
        • Peña C.
        • Silva J.
        • García V.
        • Martínez L.
        • et al.
        DeltaTAp73 upregulation correlates with poor prognosis in human tumors: putative in vivo network Involving p73 isoforms, p53, and E2F-1.
        J Clin Oncol. 2006; 24: 805-815
        • Durgin J.S.
        • Weiner D.M.
        • Wysocka M.
        • Rook A.H.
        The immunopathogenesis and immunotherapy of cutaneous T cell lymphoma: pathways and targets for immune restoration and tumor eradication’.
        J Am Acad Dermatol. 2021; 84: 587-595
        • Fiskus W.
        • Sharma S.
        • Qi J.
        • Valenta J.A.
        • Schaub L.J.
        • Shah B.
        • et al.
        Highly active combination of BRD4 antagonist and histone deacetylase inhibitor against human acute myelogenous leukemia cells.
        Mol Cancer Ther. 2014; 13: 1142-1154
        • Geng C.
        • Sang M.
        • Yang R.
        • Gao W.
        • Zhou T.
        • Wang S.
        Overexpression of 14–3-3σ counteracts tumorigenicity by positively regulating p73 in vivo.
        Oncol Lett. 2011; 2: 1177-1182
        • Greer C.B.
        • Tanaka Y.
        • Kim Y.J.
        • Xie P.
        • Zhang M.Q.
        • Park I.H.
        • et al.
        Histone deacetylases positively regulate transcription through the elongation machinery.
        Cell Rep. 2015; 13: 1444-1455
        • Halder T.G.
        • Soldi R.
        • Sharma S.
        Bromodomain and extraterminal domain protein bromodomain inhibitor based cancer therapeutics.
        Curr Opin Oncol. 2021; 33: 526-531
        • Hassan H.M.
        • Dave B.J.
        • Singh R.K.
        TP73, an under-appreciated player in non-Hodgkin lymphoma pathogenesis and management.
        Curr Mol Med. 2014; 14: 432-439
        • Jiang H.
        • Xing J.
        • Wang C.
        • Zhang H.
        • Yue L.
        • Wan X.
        • et al.
        Discovery of novel BET inhibitors by drug repurposing of nitroxoline and its analogues.
        Org Biomol Chem. 2017; 15: 9352-9361
        • Khandekar D.
        • Tiriveedhi V.
        Role of BET inhibitors in triple negative breast cancers.
        Cancers (Basel). 2020; 12: 784
        • Kiessling M.K.
        • Oberholzer P.A.
        • Mondal C.
        • Karpova M.B.
        • Zipser M.C.
        • Lin W.M.
        • et al.
        High-throughput mutation profiling of CTCL samples reveals KRAS and NRAS mutations sensitizing tumors toward inhibition of the RAS/RAF/MEK signaling cascade.
        Blood. 2011; 117: 2433-2440
        • Kim S.R.
        • Lewis J.M.
        • Cyrenne B.M.
        • Monico P.F.
        • Mirza F.N.
        • Carlson K.R.
        • et al.
        BET inhibition in advanced cutaneous T cell lymphoma is synergistically potentiated by BCL2 inhibition or HDAC inhibition.
        Oncotarget. 2018; 9: 29193-29207
        • Kopp K.L.
        • Kauczok C.S.
        • Lauenborg B.
        • Krejsgaard T.
        • Eriksen K.W.
        • Zhang Q.
        • et al.
        COX-2-dependent PGE(2) acts as a growth factor in mycosis fungoides (MF).
        Leukemia. 2010; 24: 1179-1185
        • Logotheti S.
        • Richter C.
        • Murr N.
        • Spitschak A.
        • Marquardt S.
        • Pützer B.M.
        Mechanisms of functional pleiotropy of p73 in cancer and beyond.
        Front Cell Dev Biol. 2021; 9: 737735
        • Lopez A.T.
        • Bates S.
        • Geskin L.
        Current status of HDAC inhibitors in cutaneous T-cell lymphoma.
        Am J Clin Dermatol. 2018; 19: 805-819
        • Lunghi P.
        • Costanzo A.
        • Levrero M.
        • Bonati A.
        Treatment with arsenic trioxide (ATO) and MEK1 inhibitor activates the p73-p53AIP1 apoptotic pathway in leukemia cells.
        Blood. 2004; 104: 519-525
        • Lunghi P.
        • Costanzo A.
        • Salvatore L.
        • Noguera N.
        • Mazzera L.
        • Tabilio A.
        • et al.
        MEK1 inhibition sensitizes primary acute myelogenous leukemia to arsenic trioxide–induced apoptosis.
        Blood. 2006; 107: 4549-4553
        • Müller M.
        • Schilling T.
        • Sayan A.E.
        • Kairat A.
        • Lorenz K.
        • Schulze-Bergkamen H.
        • et al.
        TAp73/DeltaNp73 influences apoptotic response, chemosensitivity and prognosis in hepatocellular carcinoma.
        Cell Death Differ. 2005; 12: 1564-1577
        • Nihal M.
        • Wu J.
        • Stonesifer C.J.
        • Daniels J.
        • Choi J.
        • Geskin L.
        • et al.
        Epigenetic regulation of apoptosis in cutaneous T-cell lymphoma: implications for therapy with methotrexate, Jak inhibitors, and resveratrol.
        J Invest Dermatol. 2022; 142: 493-496.e7
        • Park J.
        • Daniels J.
        • Wartewig T.
        • Ringbloom K.G.
        • Martinez-Escala M.E.
        • Choi S.
        • et al.
        Integrated genomic analyses of cutaneous T-cell lymphomas reveal the molecular bases for disease heterogeneity.
        Blood. 2021; 138: 1225-1236
        • Rosenbluth J.M.
        • Johnson K.
        • Tang L.
        • Triplett T.
        • Pietenpol J.A.
        Evaluation of p63 and p73 antibodies for cross-reactivity.
        Cell Cycle. 2009; 8: 3702-3706
        • Salva K.A.
        • Bennett D.
        • Longley J.
        • Guitart J.
        • Wood G.S.
        Multispectral imaging approach to the diagnosis of a CD20+ cutaneous T-cell lymphoproliferative disorder: A case report.
        Am J Dermatopathol. 2015; 37: e116-e121
        • Salva K.A.
        • Kim Y.H.
        • Rahbar Z.
        • Wood G.S.
        Epigenetically enhanced PDT induces significantly higher levels of multiple extrinsic pathway apoptotic factors than standard PDT, resulting in greater extrinsic and overall apoptosis of cutaneous T-cell lymphoma.
        Photochem Photobiol. 2018; 94: 1058-1065
        • Sayan A.E.
        • Paradisi A.
        • Vojtesek B.
        • Knight R.A.
        • Melino G.
        • Candi E.
        New antibodies recognizing p73: comparison with commercial antibodies.
        Biochem Biophys Res Commun. 2005; 330: 186-193
        • van Doorn R.
        • Zoutman W.H.
        • Dijkman R.
        • de Menezes R.X.
        • Commandeur S.
        • Mulder A.A.
        • et al.
        Epigenetic profiling of cutaneous T-cell lymphoma: promoter hypermethylation of multiple tumor suppressor genes including BCL7a, PTPRG, and p73.
        J Clin Oncol. 2005; 23: 3886-3896
        • Vilgelm A.E.
        • Hong S.M.
        • Washington M.K.
        • Wei J.
        • Chen H.
        • El-Rifai W.
        • et al.
        Characterization of ΔNp73 expression and regulation in gastric and esophageal tumors.
        Oncogene. 2010; 29: 5861-5868
        • Wakchaure P.
        • Velayutham R.
        • Roy K.K.
        Structure investigation, enrichment analysis and structure-based repurposing of FDA-approved drugs as inhibitors of BET-BRD4.
        J Biomol Struct Dyn. 2019; 37: 3048-3057
        • Weiner D.M.
        • Durgin J.S.
        • Wysocka M.
        • Rook A.H.
        The immunopathogenesis and immunotherapy of cutaneous T cell lymphoma: current and future approaches.
        J Am Acad Dermatol. 2021; 84: 597-604
        • Willemze R.
        • Cerroni L.
        • Kempf W.
        • Berti E.
        • Facchetti F.
        • Swerdlow S.H.
        • et al.
        The 2018 update of the WHO-EORTC classification for primary cutaneous lymphomas.
        Blood. 2019; 133 ([published correction appears in Blood 2019;134:1112): 1703-1714
        • Wood G.S.
        • Wu J.
        Methotrexate and pralatrexate.
        Dermatol Clin. 2015; 33: 747-755
        • Wu J.
        • Nihal M.
        • Siddiqui J.
        • Vonderheid E.C.
        • Wood G.S.
        Low FAS/CD95 expression in CTCL correlates with reduced sensitivity to apoptosis that can be restored by FAS upregulation.
        J Invest Dermatol. 2009; 129: 1165-1173
        • Wu J.
        • Wood G.S.
        Reduction of FAS/CD95 promoter methylation, upregulation of Fas protein, and enhancement of sensitivity to apoptosis in cutaneous T-cell lymphoma.
        Arch Dermatol. 2011; 147: 443-449
        • Wu J.
        • Wood G.S.
        Analysis of the effect of gentian violet on apoptosis and proliferation in cutaneous T-cell lymphoma in an in vitro study.
        JAMA Dermatol. 2018; 154: 1191-1198
        • Yoon M.K.
        • Ha J.H.
        • Lee M.S.
        • Chi S.W.
        Structure and apoptotic function of p73.
        BMB Rep. 2015; 48: 81-90
        • Yumeen S.
        • Mirza F.N.
        • Lewis J.M.
        • King A.L.O.
        • Kim S.R.
        • Carlson K.R.
        • et al.
        JAK inhibition synergistically potentiates BCL2, BET, HDAC, and proteasome inhibition in advanced CTCL.
        Blood Adv. 2020; 4: 2213-2226
        • Zhang Q.
        • Wang S.
        • Chen J.
        • Yu Z.
        Histone deacetylases (HDACs) guided novel therapies for T-cell lymphomas.
        Int J Med Sci. 2019; 16: 424-442
        • Zhang T.
        • Zhang Z.
        • Dong Q.
        • Xiong J.
        • Zhu B.
        Histone H3K27 acetylation is dispensable for enhancer activity in mouse embryonic stem cells.
        Genome Biol. 2020; 21: 45
        • Zhao L.
        • Okhovat J.P.
        • Hong E.K.
        • Kim Y.H.
        • Wood G.S.
        Preclinical studies support combined inhibition of BET family proteins and histone deacetylases as epigenetic therapy for cutaneous T-cell lymphoma.
        Neoplasia. 2019; 21: 82-92

      Supplementary References

        • Salva K.A.
        • Bennett D.
        • Longley J.
        • Guitart J.
        • Wood G.S.
        Multispectral imaging approach to the diagnosis of a CD20+ cutaneous T-cell lymphoproliferative disorder: a case report.
        Am J Dermatopathol. 2015; 37: e116-e121
        • Wu J.
        • Nihal M.
        • Siddiqui J.
        • Vonderheid E.C.
        • Wood G.S.
        Low FAS/CD95 expression in CTCL correlates with reduced sensitivity to apoptosis that can be restored by FAS upregulation.
        J Invest Dermatol. 2009; 129: 1165-1173
        • Zhao L.
        • Okhovat J.P.
        • Hong E.K.
        • Kim Y.H.
        • Wood G.S.
        Preclinical studies support combined inhibition of BET family proteins and histone deacetylases as epigenetic therapy for cutaneous T-cell lymphoma.
        Neoplasia. 2019; 21: 82-92