[en] Dysregulation of messenger RNA (mRNA) translation, including preferential translation of mRNA with complex 5' untranslated regions such as the MYC oncogene, is recognized as an important mechanism in cancer. Here, we show that both human and murine chronic lymphocytic leukemia (CLL) cells display a high translation rate, which is inhibited by the synthetic flavagline FL3, a prohibitin (PHB)-binding drug. A multiomics analysis performed in samples from patients with CLL and cell lines treated with FL3 revealed the decreased translation of the MYC oncogene and of proteins involved in cell cycle and metabolism. Furthermore, inhibiting translation induced a proliferation arrest and a rewiring of MYC-driven metabolism. Interestingly, contrary to other models, the RAS-RAF-(PHBs)-MAPK pathway is neither impaired by FL3 nor implicated in translation regulation in CLL cells. Here, we rather show that PHBs are directly associated with the eukaryotic initiation factor (eIF)4F translation complex and are targeted by FL3. Knockdown of PHBs resembled FL3 treatment. Importantly, inhibition of translation controlled CLL development in vivo, either alone or combined with immunotherapy. Finally, high expression of translation initiation-related genes and PHBs genes correlated with poor survival and unfavorable clinical parameters in patients with CLL. Overall, we demonstrated that translation inhibition is a valuable strategy to control CLL development by blocking the translation of several oncogenic pathways including MYC. We also unraveled a new and direct role of PHBs in translation initiation, thus creating new therapeutic opportunities for patients with CLL.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Largeot, Anne ; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg
Klapp, Vanessa ; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg ; Faculty of Science, Technology and Medicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg
Viry, Elodie ; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg
Gonder, Susanne; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg ; Faculty of Science, Technology and Medicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg
Fernandez Botana, Iria; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg ; Faculty of Science, Technology and Medicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg
Blomme, Arnaud ; Laboratory of Cancer Signaling, GIGA Stem Cells, University of Liège, Liège, Belgium
BENZARTI, Mohaned ; University of Luxembourg ; Department of Cancer Research, Cancer Metabolism Group, Luxembourg Institute of Health, Luxembourg, Luxembourg
Pierson, Sandrine; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg
Duculty, Chloé ; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg ; Faculty of Science, Technology and Medicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg
Marttila, Petra ; Department of Oncology-Pathology, Science for Life Laboratory, Karolinska Institutet, Solna, Sweden
Wierz, Marina; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg
Gargiulo, Ernesto ; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg
Pagano, Giulia; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg ; Faculty of Science, Technology and Medicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg
An, Ning ; Laboratory of Cancer Signaling, GIGA Stem Cells, University of Liège, Liège, Belgium
El Hachem, Najla ; Laboratory of Cancer Signaling, GIGA Stem Cells, University of Liège, Liège, Belgium
Perez Hernandez, Daniel ; Department of Infection and Immunity, Proteomics of Cellular Signaling, Luxembourg Institute of Health, Esch-sur-Alzette, Luxembourg
Chakraborty, Supriya; Molecular Hematology, International Centre for Genetic Engineering and Biotechnology, Trieste, Italy
Ysebaert, Loïc; Haematology Department, Institut Universitaire du Cancer Toulouse Oncopole, Toulouse, France
François, Jean-Hugues; Laboratoire d'hématologie, Centre Hospitalier de Luxembourg, Luxembourg, Luxembourg
Cortez Clemente, Susan; Département d'hémato-oncologie, Centre Hospitalier de Luxembourg, Luxembourg, Luxembourg
Berchem, Guy ; Département d'hémato-oncologie, Centre Hospitalier de Luxembourg, Luxembourg, Luxembourg ; Luxembourg Institute of Health, Luxembourg, Luxembourg
Efremov, Dimitar G; Molecular Hematology, International Centre for Genetic Engineering and Biotechnology, Trieste, Italy
DITTMAR, Gunnar ; University of Luxembourg ; Department of Infection and Immunity, Proteomics of Cellular Signaling, Luxembourg Institute of Health, Esch-sur-Alzette, Luxembourg
SZPAKOWSKA, Martyna ; University of Luxembourg ; Department of Infection and Immunity, Immuno-Pharmacology and Interactomics, Luxembourg Institute of Health, Esch-sur-Alzette, Luxembourg
Chevigné, Andy ; Department of Infection and Immunity, Immuno-Pharmacology and Interactomics, Luxembourg Institute of Health, Esch-sur-Alzette, Luxembourg
NAZAROV, Petr ; University of Luxembourg ; Department of Cancer Research, Multiomics Data Science, Luxembourg Institute of Health, Luxembourg, Luxembourg
Helleday, Thomas; Department of Oncology-Pathology, Science for Life Laboratory, Karolinska Institutet, Solna, Sweden ; Department of Oncology and Metabolism, Weston Park Cancer Centre, The Medical School, University of Sheffield, Sheffield, United Kingdom
Close, Pierre ; Laboratory of Cancer Signaling, GIGA Stem Cells, University of Liège, Liège, Belgium ; WELBIO Department, WEL Research Institute, Wavre, Belgium
MEISER, Johannes ; University of Luxembourg ; Department of Cancer Research, Cancer Metabolism Group, Luxembourg Institute of Health, Luxembourg, Luxembourg
Stamatopoulos, Basile; Laboratory of Clinical Cell Therapy, ULB-Research Cancer Center, Jules Bordet Institute, Université Libre de Bruxelles, Brussels, Belgium
Désaubry, Laurent ; Regenerative Nanomedicine Laboratory (UMR1260), Faculty of Medicine, Fédération de Médecine Translationnelle de Strasbourg, INSERM-University of Strasbourg, Strasbourg, France
PAGGETTI, Jerome ✱; University of Luxembourg ; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg
MOUSSAY, Etienne ✱; University of Luxembourg ; Department of Cancer Research, Tumor Stroma Interactions, Luxembourg Institute of Health, Luxembourg, Luxembourg
The authors thank Carlo Croce and John Byrd (Ohio State University, US) for the kind gift of Eμ-TCL1 mouse and the OSU-CLL cell line. The authors also thank the National Cytometry Platform (LIH; Antonio Cosma, Céline Hoffmann, Thomas Cerutti, Fanny Hedin, Mario Gomez) for assistance in flow cytometry and confocal microscopy, the bioinformatics platform (LIH; Reka Toth) for assistance, and the animal facility (LIH) staff, particularly Anais Oudin and Coralie Pulido; the Metabolomics Platform (LIH, Antoine Lesur, François Bernardin) for liquid chromatography mass spectrometry measurements and for providing technical and analytical support; the LUXGEN platform (LIH/LNS; Nathalie Nicot, Pol Hoffmann, Arnaud Muller and Daniel Stieber) for RNA sequencing; and the GIGA genomics facility. The authors also thank Nadia Beaupain, Jean-Marc Plesseria, and Manuel Counson from the Immuno-Pharmacology and Interactomics group (LIH) for their help in the NanoBRET experiments; Vincent Schlesser, Sigrid De Wilde, Laurent Plawny, and Sebastien Rinaldetti from the Centre Hospitalier du Luxembourg for their help in sample collection; and Titiksha Basu, Jil Delmarque, Olinda Pinto, Carmen Lahr, and Audrey Kopp for technical support. Finally, the authors thank Maxmilan Jeyakumar (LIH) for proofreading the manuscript. This work was supported by grants from the Luxembourg National Research Fund (FNR) and Fondation Cancer to V.K. E.G. C.D. E.M. and J.P. (PRIDE19/14254520/i2TRON, PRIDE15/10675146/CANBIO, PRIDE21/16763386, C20/BM/14582635, and C20/BM/14592342); to A.C. and M.S. (INTER/FNRS/20/15084569); and to J.M. (ATTRACT grant A18/BM/11809970); from FNRS-Télévie to A.L. (7.4502.17, 7.4503.19), E.V. (7.4509.20, 7.4572.22), S.G. (7.4502.19, 7.6604.21), G.P. (7.4501.18, 7.6518.20), I.F.B. (7.4529.19, 7.6603.21), M.W. (7.4508.16, 7.6504.18), A.C. and M.S. (7.8508.22, 7.8504.20 and 7.4593.19); from the Plooschter Projet to J.P. and E.M.; from the Belgian Foundation for Cancer Research to P.C. (No 2020-068); and from the Swedish Children's Cancer Foundation (PR2021-003), the Swedish Research Council (2015-00162), and Swedish Cancer Society (21 1490) to T.H. Contribution: A.L. designed and performed experiments, analyzed results, and wrote the manuscript; V.K. E.V. S.G. I.F.B. S.P. C.D. M.W. E.G. and G.P. performed experiments and analyzed data; A.B. N.A. N.E.H. and P.C. performed L-propargylglycine assay, polysome profiling and analysis, and provided expertise on translation; P.M. and T.H. performed DARTS assay and analysis; D.P.H. and G.D. performed pulsed SILAC experiments and analysis; S.C. and D.G.E. provided materials and expertise; P.V.N. performed statistical analysis of gene expression data in patients with CLL; M.B. and J.M. performed metabolomics experiments and analyzed data; M.S. and A.C. performed BRET experiments and analyzed data; B.S. provided sample from patients with CLL, complementary DNA from cohort of patients with CLL, patients’ data, and expertise for analyses; L.D. synthetized FL3 and provided his expertise on the molecule; L.Y. J.-H.F. S.C.C. and G.B. provided samples from patients with CLL, and expertise on hemato-oncology; E.M. and J.P. designed and supervised the study, performed bioinformatics analyses, analyzed results, and wrote the final version of the manuscript; and all authors revised the manuscript.This work was supported by grants from the Luxembourg National Research Fund (FNR) and Fondation Cancer to V.K., E.G., C.D., E.M., and J.P. (PRIDE19/14254520/i2TRON, PRIDE15/10675146/CANBIO, PRIDE21/16763386, C20/BM/14582635, and C20/BM/14592342); to A.C. and M.S. (INTER/FNRS/20/15084569); and to J.M. (ATTRACT grant A18/BM/11809970); from FNRS-Télévie to A.L. (7.4502.17, 7.4503.19), E.V. (7.4509.20, 7.4572.22), S.G. (7.4502.19, 7.6604.21), G.P. (7.4501.18, 7.6518.20), I.F.B. (7.4529.19, 7.6603.21), M.W. (7.4508.16, 7.6504.18), A.C. and M.S. (7.8508.22, 7.8504.20 and 7.4593.19); from the Plooschter Projet to J.P. and E.M.; from the Belgian Foundation for Cancer Research to P.C. (No 2020-068); and from the Swedish Children’s Cancer Foundation (PR2021-003), the Swedish Research Council (2015-00162), and Swedish Cancer Society (21 1490) to T.H.
Burger, JA, Chiorazzi, N, B cell receptor signaling in chronic lymphocytic leukemia. Trends Immunol 34:12 (2013), 592–601.
Dadashian, EL, McAuley, EM, Liu, D, et al. TLR signaling is activated in lymph node-resident CLL cells and is only partially inhibited by ibrutinib. Cancer Res 79:2 (2019), 360–371.
Brown, JR, How I treat CLL patients with ibrutinib. Blood 131:4 (2018), 379–386.
Filip, D, Mraz, M, The role of MYC in the transformation and aggressiveness of 'indolent' B-cell malignancies. Leuk Lymphoma 61:3 (2020), 510–524.
Moussay, E, Palissot, V, Vallar, L, et al. Determination of genes and microRNAs involved in the resistance to fludarabine in vivo in chronic lymphocytic leukemia. Mol Cancer, 9, 2010, 115.
Nguyen-Khac, F, “Double-hit” chronic lymphocytic leukemia, involving the TP53 and MYC genes. Front Oncol, 11, 2021, 826245.
Chen, Z, Simon-Molas, H, Cretenet, G, et al. Characterization of metabolic alterations of chronic lymphocytic leukemia in the lymph node microenvironment. Blood 140:6 (2022), 630–643.
Herishanu, Y, Perez-Galan, P, Liu, D, et al. The lymph node microenvironment promotes B-cell receptor signaling, NF-kappaB activation, and tumor proliferation in chronic lymphocytic leukemia. Blood 117:2 (2011), 563–574.
Krysov, S, Dias, S, Paterson, A, et al. Surface IgM stimulation induces MEK1/2-dependent MYC expression in chronic lymphocytic leukemia cells. Blood 119:1 (2012), 170–179.
Yeomans, A, Thirdborough, SM, Valle-Argos, B, et al. Engagement of the B-cell receptor of chronic lymphocytic leukemia cells drives global and MYC-specific mRNA translation. Blood 127:4 (2016), 449–457.
Ruggero, D, Translational control in cancer etiology. Cold Spring Harb Perspect Biol, 5(2), 2013, a012336.
Silvera, D, Formenti, SC, Schneider, RJ, Translational control in cancer. Nat Rev Cancer 10:4 (2010), 254–266.
Boussemart, L, Malka-Mahieu, H, Girault, I, et al. eIF4F is a nexus of resistance to anti-BRAF and anti-MEK cancer therapies. Nature 513:7516 (2014), 105–109.
Bretones, G, Alvarez, MG, Arango, JR, et al. Altered patterns of global protein synthesis and translational fidelity in RPS15-mutated chronic lymphocytic leukemia. Blood 132:22 (2018), 2375–2388.
Ljungstrom, V, Cortese, D, Young, E, et al. Whole-exome sequencing in relapsing chronic lymphocytic leukemia: clinical impact of recurrent RPS15 mutations. Blood 127:8 (2016), 1007–1016.
Ntoufa, S, Gerousi, M, Laidou, S, et al. RPS15 mutations rewire RNA translation in chronic lymphocytic leukemia. Blood Adv 5:13 (2021), 2788–2792.
Sbarrato, T, Horvilleur, E, Poyry, T, et al. A ribosome-related signature in peripheral blood CLL B cells is linked to reduced survival following treatment. Cell Death Dis, 7(6), 2016, e2249.
Paggetti, J, Moussay, E, BCR engagement in CLL: when translation goes wrong. Blood 127:4 (2016), 378–380.
Chen, M, Asanuma, M, Takahashi, M, et al. Dual targeting of DDX3 and eIF4A by the translation inhibitor rocaglamide A. Cell Chem Biol 28:4 (2021), 475–486.e8.
Chen, J, Sathiaseelan, V, Moore, A, et al. ZAP-70 constitutively regulates gene expression and protein synthesis in chronic lymphocytic leukemia. Blood 137:26 (2021), 3629–3640.
Willimott, S, Beck, D, Ahearne, MJ, Adams, VC, Wagner, SD, Cap-translation inhibitor, 4EGI-1, restores sensitivity to ABT-737 apoptosis through cap-dependent and -independent mechanisms in chronic lymphocytic leukemia. Clin Cancer Res 19:12 (2013), 3212–3223.
Wilmore, S, Rogers-Broadway, KR, Taylor, J, et al. Targeted inhibition of eIF4A suppresses B-cell receptor-induced translation and expression of MYC and MCL1 in chronic lymphocytic leukemia cells. Cell Mol Life Sci 78:17-18 (2021), 6337–6349.
Cencic, R, Carrier, M, Galicia-Vazquez, G, et al. Antitumor activity and mechanism of action of the cyclopenta[b]benzofuran, silvestrol. PLoS One, 4(4), 2009, e5223.
Jackson, DN, Alula, KM, Delgado-Deida, Y, et al. The synthetic small molecule FL3 combats intestinal tumorigenesis via Axin1-mediated inhibition of Wnt/beta-catenin signaling. Cancer Res 80:17 (2020), 3519–3529.
Basmadjian, C, Thuaud, F, Ribeiro, N, Desaubry, L, Flavaglines: potent anticancer drugs that target prohibitins and the helicase eIF4A. Future Med Chem 5:18 (2013), 2185–2197.
Nebigil, CG, Moog, C, Vagner, S, Benkirane-Jessel, N, Smith, DR, Desaubry, L, Flavaglines as natural products targeting eIF4A and prohibitins: from traditional Chinese medicine to antiviral activity against coronaviruses. Eur J Med Chem, 203, 2020, 112653.
Thuaud, F, Ribeiro, N, Nebigil, CG, Desaubry, L, Prohibitin ligands in cell death and survival: mode of action and therapeutic potential. Chem Biol 20:3 (2013), 316–331.
Bichi, R, Shinton, SA, Martin, ES, et al. Human chronic lymphocytic leukemia modeled in mouse by targeted TCL1 expression. Proc Natl Acad Sci U S A 99:10 (2002), 6955–6960.
Chakraborty, S, Martines, C, Porro, F, et al. B-cell receptor signaling and genetic lesions in TP53 and CDKN2A/CDKN2B cooperate in Richter transformation. Blood 138:12 (2021), 1053–1066.
Ho, JJD, Cunningham, TA, Manara, P, et al. Proteomics reveal cap-dependent translation inhibitors remodel the translation machinery and translatome. Cell Rep, 37(2), 2021, 109806.
Tambay, V, Raymond, VA, Bilodeau, M, MYC rules: leading glutamine metabolism toward a distinct cancer cell phenotype. Cancers, 13(17), 2021, 4484.
Wise, DR, DeBerardinis, RJ, Mancuso, A, et al. Myc regulates a transcriptional program that stimulates mitochondrial glutaminolysis and leads to glutamine addiction. Proc Natl Acad Sci U S A 105:48 (2008), 18782–18787.
Rajalingam, K, Wunder, C, Brinkmann, V, et al. Prohibitin is required for Ras-induced Raf-MEK-ERK activation and epithelial cell migration. Nat Cell Biol 7:8 (2005), 837–843.
Waskiewicz, AJ, Flynn, A, Proud, CG, Cooper, JA, Mitogen-activated protein kinases activate the serine/threonine kinases Mnk1 and Mnk2. EMBO J 16:8 (1997), 1909–1920.
Avdulov, S, Li, S, Michalek, V, et al. Activation of translation complex eIF4F is essential for the genesis and maintenance of the malignant phenotype in human mammary epithelial cells. Cancer Cell 5:6 (2004), 553–563.
Yang, M, Yao, B, Lin, R, Jin, H, The translational regulation in mTOR pathway. Biomolecules, 12(12), 2022, 1790.
Iwasaki, S, Iwasaki, W, Takahashi, M, et al. The translation inhibitor rocaglamide targets a bimolecular cavity between eIF4A and polypurine RNA. Mol Cell 73:4 (2019), 738–748.e9.
Kampen, KR, Sulima, SO, Vereecke, S, De Keersmaecker, K, Hallmarks of ribosomopathies. Nucleic Acids Res 48:3 (2020), 1013–1028.
Rasul, E, Salamon, D, Nagy, N, et al. The MEC1 and MEC2 lines represent two CLL subclones in different stages of progression towards prolymphocytic leukemia. PLoS One, 9(8), 2014, e106008.
Wolfe, AL, Singh, K, Zhong, Y, et al. RNA G-quadruplexes cause eIF4A-dependent oncogene translation in cancer. Nature 513:7516 (2014), 65–70.
Taylor, J, Yeomans, AM, Packham, G, Targeted inhibition of mRNA translation initiation factors as a novel therapeutic strategy for mature B-cell neoplasms. Explor Target Antitumor Ther 1:1 (2020), 3–25.
Polier, G, Neumann, J, Thuaud, F, et al. The natural anticancer compounds rocaglamides inhibit the Raf-MEK-ERK pathway by targeting prohibitin 1 and 2. Chem Biol 19:9 (2012), 1093–1104.
Sadlish, H, Galicia-Vazquez, G, Paris, CG, et al. Evidence for a functionally relevant rocaglamide binding site on the eIF4A-RNA complex. ACS Chem Biol 8:7 (2013), 1519–1527.
Bordeleau, ME, Robert, F, Gerard, B, et al. Therapeutic suppression of translation initiation modulates chemosensitivity in a mouse lymphoma model. J Clin Invest 118:7 (2008), 2651–2660.
Chiu, CF, Ho, MY, Peng, JM, et al. Raf activation by Ras and promotion of cellular metastasis require phosphorylation of prohibitin in the raft domain of the plasma membrane. Oncogene 32:6 (2013), 777–787.
Mishra, S, Ande, SR, Nyomba, BL, The role of prohibitin in cell signaling. FEBS J 277:19 (2010), 3937–3946.
Furic, L, Rong, L, Larsson, O, et al. eIF4E phosphorylation promotes tumorigenesis and is associated with prostate cancer progression. Proc Natl Acad Sci U S A 107:32 (2010), 14134–14139.
Ueda, T, Sasaki, M, Elia, AJ, et al. Combined deficiency for MAP kinase-interacting kinase 1 and 2 (Mnk1 and Mnk2) delays tumor development. Proc Natl Acad Sci U S A 107:32 (2010), 13984–13990.
Wendel, HG, Silva, RL, Malina, A, et al. Dissecting eIF4E action in tumorigenesis. Genes Dev 21:24 (2007), 3232–3237.
Ueda, T, Watanabe-Fukunaga, R, Fukuyama, H, Nagata, S, Fukunaga, R, Mnk2 and Mnk1 are essential for constitutive and inducible phosphorylation of eukaryotic initiation factor 4E but not for cell growth or development. Mol Cell Biol 24:15 (2004), 6539–6549.
Shveygert, M, Kaiser, C, Bradrick, SS, Gromeier, M, Regulation of eukaryotic initiation factor 4E (eIF4E) phosphorylation by mitogen-activated protein kinase occurs through modulation of Mnk1-eIF4G interaction. Mol Cell Biol 30:21 (2010), 5160–5167.
Wierz, M, Janji, B, Berchem, G, Moussay, E, Paggetti, J, High-dimensional mass cytometry analysis revealed microenvironment complexity in chronic lymphocytic leukemia. Oncoimmunology, 7(8), 2018, e1465167.
Wierz, M, Pierson, S, Guyonnet, L, et al. Dual PD1/LAG3 immune checkpoint blockade limits tumor development in a murine model of chronic lymphocytic leukemia. Blood 131:14 (2018), 1617–1621.
Volta, V, Perez-Baos, S, de la Parra, C, et al. A DAP5/eIF3d alternate mRNA translation mechanism promotes differentiation and immune suppression by human regulatory T cells. Nat Commun, 12(1), 2021, 6979.
Zhu, X, Zhang, W, Guo, J, et al. Noc4L-mediated ribosome biogenesis controls activation of regulatory and conventional T cells. Cell Rep 27:4 (2019), 1205–1220.e4.
