{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T00:51:18Z","timestamp":1760057478980,"version":"build-2065373602"},"reference-count":33,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,2,5]],"date-time":"2025-02-05T00:00:00Z","timestamp":1738713600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Instituto de Investigaci\u00f3n Marqu\u00e9s de Valdecilla (IDIVAL)","award":["TRANSVAL20\/03"],"award-info":[{"award-number":["TRANSVAL20\/03"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Background\/Objectives: Chronic lymphocytic leukemia (CLL) remains an incurable B-cell malignancy. B-CLL cells exhibit an extended lifespan in part due to the activation of survival pathways such as NF-kB. A crosstalk between NF-kB and GSK-3\u03b2 pathways has been reported. NF-kB has also been identified as a primary target of the NEDD8-activating enzyme inhibitor MLN4924. Our objective was to investigate potential synergies of MLN4924 with other NF-kB-targeting agents for the treatment of CLL and elucidate the mechanisms of action underlying this pathway regulation. Methods: To assess the cytotoxic efficacy of the combined ex vivo treatment with CHIR-99021 and MLN4924, we employed 7-AAD staining and XTT viability assays on primary samples from CLL patients. Subsequently, we conducted various analyses to identify the molecular mechanisms underlying the cytotoxic effects of this combination. Results: We discovered a discrepancy between the mRNA and protein levels of IkB\u0251 and provided evidence of translational control over its expression. This observation may explain why, unlike other cell types, B-CLL cells did not activate NF-kB signaling following inhibition of GSK-3\u00df. Furthermore, we describe a synergistic effect between a specific GSK-3\u00df inhibitor, CHIR-99021\/Laduviglusib, and the NEDD8-activating enzyme inhibitor MLN4924\/Pevonedistat, at doses that only slightly affect healthy B cell viability ex vivo. We investigated the molecular basis of this co-induction of cell death by analyzing the alterations in apoptosis-related gene expression. We found that the combinational treatment enhances a reduction in BCL2 mRNA expression levels, providing an alternative approach for BCL-2 inhibition in CLL that could have therapeutic implications for the treatment of refractory CLL cases. Conclusions: our findings revealed a unique interaction between GSK-3\u00df and NF-kB pathways in CLL and their regulation of BCL2 expression.<\/jats:p>","DOI":"10.3390\/cancers17030533","type":"journal-article","created":{"date-parts":[[2025,2,5]],"date-time":"2025-02-05T05:50:36Z","timestamp":1738734636000},"page":"533","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Distinct NF-kB Regulation Favors a Synergic Action of Pevonedistat and Laduviglusib in B-Chronic Lymphocytic Leukemia Cells Ex Vivo"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4548-6422","authenticated-orcid":false,"given":"V\u00edctor","family":"Arenas","sequence":"first","affiliation":[{"name":"Molecular Hematology Laboratory, Marqu\u00e9s de Valdecilla Research Institute, 39008 Santander, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7670-5515","authenticated-orcid":false,"given":"Jose Luis","family":"Casta\u00f1o","sequence":"additional","affiliation":[{"name":"Molecular Hematology Laboratory, Marqu\u00e9s de Valdecilla Research Institute, 39008 Santander, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juan Jos\u00e9","family":"Dom\u00ednguez","sequence":"additional","affiliation":[{"name":"Hematology Department, Marqu\u00e9s de Valdecilla University Hospital, 39008 Santander, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lucrecia","family":"Y\u00e1\u00f1ez","sequence":"additional","affiliation":[{"name":"Hematology Department, Marqu\u00e9s de Valdecilla University Hospital, 39008 Santander, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3851-6710","authenticated-orcid":false,"given":"Carlos","family":"Pipa\u00f3n","sequence":"additional","affiliation":[{"name":"Molecular Hematology Laboratory, Marqu\u00e9s de Valdecilla Research Institute, 39008 Santander, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"16096","DOI":"10.1038\/nrdp.2016.96","article-title":"Chronic lymphocytic leukaemia","volume":"3","author":"Kipps","year":"2017","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"684","DOI":"10.1038\/s41571-019-0239-8","article-title":"Chronic lymphocytic leukaemia: From genetics to treatment","volume":"16","author":"Bosch","year":"2019","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.semcancer.2016.07.005","article-title":"NF-\u03baB activation in chronic lymphocytic leukemia: A point of convergence of external triggers and intrinsic lesions","volume":"39","author":"Mansouri","year":"2016","journal-title":"Semin. Cancer Biol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Oeckinghaus, A., and Ghosh, S. (2009). The NF-kappaB family of transcription factors and its regulation. Cold Spring Harb. Perspect. Biol., 1.","DOI":"10.1101\/cshperspect.a000034"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1038\/ni.2065","article-title":"Crosstalk in NF-\u03baB signaling pathways","volume":"12","author":"Oeckinghaus","year":"2011","journal-title":"Nat. Immunol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"49571","DOI":"10.1074\/jbc.C400442200","article-title":"Phosphorylation of serine 468 by GSK-3beta negatively regulates basal p65 NF-kappaB activity","volume":"279","author":"Buss","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.cell.2009.01.038","article-title":"A Nurr1\/CoREST pathway in microglia and astrocytes protects dopaminergic neurons from inflammation-induced death","volume":"137","author":"Saijo","year":"2009","journal-title":"Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1182\/blood-2006-12-060947","article-title":"Inhibition of glycogen synthase kinase-3 activity leads to epigenetic silencing of nuclear factor kappaB target genes and induction of apoptosis in chronic lymphocytic leukemia B cells","volume":"110","author":"Ougolkov","year":"2007","journal-title":"Blood"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2619","DOI":"10.1042\/BCJ20210139","article-title":"Inhibitory feedback control of NF-\u03baB signalling in health and disease","volume":"478","author":"Prescott","year":"2021","journal-title":"Biochem. J."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1515","DOI":"10.1182\/blood-2010-03-272567","article-title":"MLN4924, a NEDD8-activating enzyme inhibitor, is active in diffuse large B-cell lymphoma models: Rationale for treatment of NF-{kappa}B-dependent lymphoma","volume":"116","author":"Milhollen","year":"2010","journal-title":"Blood"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1576","DOI":"10.1158\/1078-0432.CCR-13-0987","article-title":"The Nedd8-activating enzyme inhibitor MLN4924 thwarts microenvironment-driven NF-\u03baB activation and induces apoptosis in chronic lymphocytic leukemia B cells","volume":"20","author":"Godbersen","year":"2014","journal-title":"Clin. Cancer Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1038\/s41408-022-00763-w","article-title":"Pevonedistat, a Nedd8-activating enzyme inhibitor, in combination with ibrutinib in patients with relapsed\/refractory B-cell non-Hodgkin lymphoma","volume":"13","author":"Torka","year":"2023","journal-title":"Blood Cancer J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3568","DOI":"10.1038\/s41375-021-01184-7","article-title":"Map of ubiquitin-like post-translational modifications in chronic lymphocytic leukemia. Role of p53 lysine 120 NEDDylation","volume":"35","author":"Briz","year":"2021","journal-title":"Leukemia"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Arenas, V., Casta\u00f1o, J.L., Dom\u00ednguez-Garc\u00eda, J.J., Y\u00e1\u00f1ez, L., and Pipa\u00f3n, C. (2021). A Different View for an Old Disease: NEDDylation and Other Ubiquitin-Like Post-Translational Modifications in Chronic Lymphocytic Leukemia. Front. Oncol., 11.","DOI":"10.3389\/fonc.2021.729550"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1038\/ni.2291","article-title":"Translational control of the activation of transcription factor NF-\u03baB and production of type I interferon by phosphorylation of the translation factor eIF4E","volume":"13","author":"Herdy","year":"2012","journal-title":"Nat. Immunol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2778","DOI":"10.1038\/s41388-022-02272-3","article-title":"Epigenetic regulation of EIF4A1 through DNA methylation and an oncogenic role of eIF4A1 through BRD2 signaling in prostate cancer","volume":"41","author":"Wang","year":"2022","journal-title":"Oncogene"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Malina, A., Mills, J.R., and Pelletier, J. (2012). Emerging therapeutics targeting mRNA translation. Cold Spring Harb. Perspect. Biol., 4.","DOI":"10.1101\/cshperspect.a012377"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6337","DOI":"10.1007\/s00018-021-03910-x","article-title":"Targeted inhibition of eIF4A suppresses B-cell receptor-induced translation and expression of MYC and MCL1 in chronic lymphocytic leukemia cells","volume":"78","author":"Wilmore","year":"2021","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1038\/s41375-019-0503-z","article-title":"Targeting translation initiation by synthetic rocaglates for treating MYC-driven lymphomas","volume":"34","author":"Zhang","year":"2020","journal-title":"Leukemia"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1566","DOI":"10.3109\/10428194.2014.990901","article-title":"Targeting neddylation effectively antagonizes nuclear factor-\u03baB in chronic lymphocytic leukemia B-cells","volume":"56","author":"Godbersen","year":"2015","journal-title":"Leuk. Lymphoma"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2469","DOI":"10.1038\/s41375-021-01308-z","article-title":"Inhibition of translation initiation factor eIF4a inactivates heat shock factor 1 (HSF1) and exerts anti-leukemia activity in AML","volume":"35","author":"Nishida","year":"2021","journal-title":"Leukemia"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Martelli, A.M., Paganelli, F., Evangelisti, C., Chiarini, F., and McCubrey, J.A. (2022). Pathobiology and Therapeutic Relevance of GSK-3 in Chronic Hematological Malignancies. Cells, 11.","DOI":"10.3390\/cells11111812"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1016\/j.gpb.2022.01.004","article-title":"SynergyFinder Plus: Toward Better Interpretation and Annotation of Drug Combination Screening Datasets","volume":"20","author":"Zheng","year":"2022","journal-title":"Genom. Proteom. Bioinform."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1084\/jem.20142009","article-title":"Functional loss of I\u03baB\u03b5 leads to NF-\u03baB deregulation in aggressive chronic lymphocytic leukemia","volume":"212","author":"Mansouri","year":"2015","journal-title":"J. Exp. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1182\/blood-2015-07-660969","article-title":"Engagement of the B-cell receptor of chronic lymphocytic leukemia cells drives global and MYC-specific mRNA translation","volume":"127","author":"Yeomans","year":"2016","journal-title":"Blood"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"4694","DOI":"10.18632\/oncotarget.2100","article-title":"High p27 protein levels in chronic lymphocytic leukemia are associated to low Myc and Skp2 expression, confer resistance to apoptosis and antagonize Myc effects on cell cycle","volume":"5","author":"Caraballo","year":"2014","journal-title":"Oncotarget"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1373","DOI":"10.1002\/cac2.12493","article-title":"Targeting mRNA translation aberrations: A novel approach for therapy in chronic lymphocytic leukemia","volume":"43","author":"Klapp","year":"2023","journal-title":"Cancer Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4269","DOI":"10.1038\/onc.2015.493","article-title":"Oncogenic CARMA1 couples NF-\u03baB and \u03b2-catenin signaling in diffuse large B-cell lymphomas","volume":"35","author":"Bognar","year":"2016","journal-title":"Oncogene"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2934","DOI":"10.1038\/s41388-020-1183-x","article-title":"Stabilization of \u03b2-catenin upon B-cell receptor signaling promotes NF-kB target genes transcription in mantle cell lymphoma","volume":"39","author":"Lazarian","year":"2020","journal-title":"Oncogene"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3118","DOI":"10.1073\/pnas.0308648100","article-title":"Activation of the Wnt signaling pathway in chronic lymphocytic leukemia","volume":"101","author":"Lu","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1038\/s41419-018-1200-y","article-title":"Cul4 E3 ubiquitin ligase regulates ovarian cancer drug resistance by targeting the antiapoptotic protein BIRC3","volume":"10","author":"Hu","year":"2019","journal-title":"Cell Death Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2122","DOI":"10.1182\/blood-2015-07-659144","article-title":"Clinical impact of clonal and subclonal TP53, SF3B1, BIRC3, NOTCH1, and ATM mutations in chronic lymphocytic leukemia","volume":"127","author":"Nadeu","year":"2016","journal-title":"Blood"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1901","DOI":"10.1158\/1078-0432.CCR-18-1548","article-title":"BIRC3 Expression Predicts CLL Progression and Defines Treatment Sensitivity via Enhanced NF-\u03baB Nuclear Translocation","volume":"25","author":"Asslaber","year":"2019","journal-title":"Clin. Cancer Res."}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/17\/3\/533\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:27:07Z","timestamp":1760027227000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/17\/3\/533"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,5]]},"references-count":33,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["cancers17030533"],"URL":"https:\/\/doi.org\/10.3390\/cancers17030533","relation":{},"ISSN":["2072-6694"],"issn-type":[{"type":"electronic","value":"2072-6694"}],"subject":[],"published":{"date-parts":[[2025,2,5]]}}}