{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:39:02Z","timestamp":1787027942330,"version":"build-2736575974"},"reference-count":55,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,1,8]],"date-time":"2025-01-08T00:00:00Z","timestamp":1736294400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"All Japan Coffee Association","award":["2244"],"award-info":[{"award-number":["2244"]}]},{"name":"Salt Science Research Foundation","award":["2244"],"award-info":[{"award-number":["2244"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diseases"],"abstract":"<jats:p>Background: Acute myeloid leukemia (AML) is a common and aggressive form of leukemia, yet current treatment strategies remain insufficient. Venetoclax, a BH3-mimetic approved for AML treatment, induces Bcl-2-dependent apoptosis, though its therapeutic efficacy is still limited. Therefore, new strategies to enhance the effect of venetoclax are highly sought. Valproic acid (VPA), commonly used for epilepsy, has also been studied for potential applications in AML treatment. Methods: AML cells were treated with venetoclax, with or without VPA. Cell viability was assessed using the trypan blue dye exclusion assay, while cell cycle progression was analyzed by flow cytometry. The expression of pro-apoptotic proteins Bax and Bak was measured by RT-qPCR. Results: Venetoclax and VPA individually had only mild effects on AML cell proliferation. However, their combination significantly inhibited cell growth and triggered pronounced cell death. This combination also led to the cleavage of poly (ADP-ribose) polymerase (PARP), a substrate of caspases, indicating activation of apoptosis. VPA treatment upregulated the expression of Bax and Bak, further supporting apoptosis induction. The cell death induced by the venetoclax\u2013VPA combination was predominantly apoptotic, as confirmed by the near-complete blockade of cell death by a pan-caspase inhibitor. Conclusions: Our study demonstrates that VPA enhances venetoclax-induced apoptosis in AML cell lines, providing a novel role for VPA and suggesting a promising combinatory strategy for AML treatment. These findings offer valuable insights into potential clinical applications of venetoclax and VPA in AML management.<\/jats:p>","DOI":"10.3390\/diseases13010010","type":"journal-article","created":{"date-parts":[[2025,1,8]],"date-time":"2025-01-08T11:56:22Z","timestamp":1736337382000},"page":"10","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Valproic Acid Enhances Venetoclax Efficacy in Targeting Acute Myeloid Leukemia"],"prefix":"10.3390","volume":"13","author":[{"given":"Renshi","family":"Kawakatsu","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenjiro","family":"Tadagaki","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenta","family":"Yamasaki","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasumichi","family":"Kuwahara","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan"},{"name":"Department of Kyoto Pediatric Community-Based Medicine, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0634-5160","authenticated-orcid":false,"given":"Tatsushi","family":"Yoshida","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1016\/S0140-6736(18)31041-9","article-title":"Acute myeloid leukaemia","volume":"392","author":"Short","year":"2018","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"De Kouchkovsky, I., and Abdul-Hay, M. (2016). Acute myeloid leukemia: A comprehensive review and 2016 update. Blood Cancer J., 6.","DOI":"10.1038\/bcj.2016.50"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Boffo, S., Damato, A., Alfano, L., and Giordano, A. (2018). CDK9 inhibitors in acute myeloid leukemia. J. Exp. Clin. Cancer Res., 37.","DOI":"10.1186\/s13046-018-0704-8"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"4422","DOI":"10.1182\/blood-2010-03-276485","article-title":"Intensive chemotherapy does not benefit most older patients (age 70 years or older) with acute myeloid leukemia","volume":"116","author":"Kantarjian","year":"2010","journal-title":"Blood"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Pettit, K., and Odenike, O. (2015). Defining and Treating Older Adults with Acute Myeloid Leukemia Who Are Ineligible for Intensive Therapies. Front. Oncol., 5.","DOI":"10.3389\/fonc.2015.00280"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1182\/blood-2015-01-621664","article-title":"International phase 3 study of azacitidine vs conventional care regimens in older patients with newly diagnosed AML with >30% blasts","volume":"126","author":"Dombret","year":"2015","journal-title":"Blood"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Hafezi, S., and Rahmani, M. (2021). Targeting BCL-2 in Cancer: Advances, Challenges, and Perspectives. Cancers, 13.","DOI":"10.3390\/cancers13061292"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"5750","DOI":"10.1021\/acs.jmedchem.8b01668","article-title":"Inhibitor of Apoptosis Protein (IAP) Antagonists in Anticancer Agent Discovery: Current Status and Perspectives","volume":"62","author":"Cong","year":"2019","journal-title":"J. Med. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.semcdb.2014.07.002","article-title":"Therapeutic exploitation of necroptosis for cancer therapy","volume":"35","author":"Fulda","year":"2014","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1016\/j.ceb.2004.09.011","article-title":"Death by design: Apoptosis, necrosis and autophagy","volume":"16","author":"Edinger","year":"2004","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3145","DOI":"10.3390\/ijms15023145","article-title":"Autophagic cell death and cancer","volume":"15","author":"Shimizu","year":"2014","journal-title":"Int. J. Mol. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"700","DOI":"10.1038\/nrm2970","article-title":"Molecular mechanisms of necroptosis: An ordered cellular explosion","volume":"11","author":"Vandenabeele","year":"2010","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1060","DOI":"10.1016\/j.cell.2012.03.042","article-title":"Ferroptosis: An iron-dependent form of nonapoptotic cell death","volume":"149","author":"Dixon","year":"2012","journal-title":"Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.tibs.2016.10.004","article-title":"Pyroptosis: Gasdermin-Mediated Programmed Necrotic Cell Death","volume":"42","author":"Shi","year":"2017","journal-title":"Trends Biochem. Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"147","DOI":"10.4161\/cbt.4.2.1508","article-title":"Overview of cell death signaling pathways","volume":"4","author":"Jin","year":"2005","journal-title":"Cancer Biol. Ther."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1031","DOI":"10.1038\/sj.cdd.4401088","article-title":"The role of mitochondrial factors in apoptosis: A Russian roulette with more than one bullet","volume":"9","author":"Saelens","year":"2002","journal-title":"Cell Death Differ."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1152\/physrev.00013.2006","article-title":"Mitochondrial membrane permeabilization in cell death","volume":"87","author":"Kroemer","year":"2007","journal-title":"Physiol. Rev."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1016\/S0014-5793(99)01761-5","article-title":"Bcl-2 family: Life-or-death switch","volume":"466","author":"Tsujimoto","year":"2000","journal-title":"FEBS Lett."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1200\/JCO.2010.31.6208","article-title":"Phase I study of Navitoclax (ABT-263), a novel Bcl-2 family inhibitor, in patients with small-cell lung cancer and other solid tumors","volume":"29","author":"Gandhi","year":"2011","journal-title":"J. Clin. Oncol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3163","DOI":"10.1158\/1078-0432.CCR-11-3090","article-title":"Phase II study of single-agent Navitoclax (ABT-263) and biomarker correlates in patients with relapsed small cell lung cancer","volume":"18","author":"Rudin","year":"2012","journal-title":"Clin. Cancer Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1149","DOI":"10.1016\/S1470-2045(10)70261-8","article-title":"Navitoclax, a targeted high-affinity inhibitor of BCL-2, in lymphoid malignancies: A phase 1 dose-escalation study of safety, pharmacokinetics, pharmacodynamics, and antitumour activity","volume":"11","author":"Wilson","year":"2010","journal-title":"Lancet Oncol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"488","DOI":"10.1200\/JCO.2011.34.7898","article-title":"Substantial susceptibility of chronic lymphocytic leukemia to BCL2 inhibition: Results of a phase I study of navitoclax in patients with relapsed or refractory disease","volume":"30","author":"Roberts","year":"2012","journal-title":"J. Clin. Oncol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1038\/nm.3048","article-title":"ABT-199, a potent and selective BCL-2 inhibitor, achieves antitumor activity while sparing platelets","volume":"19","author":"Souers","year":"2013","journal-title":"Nat. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4042","DOI":"10.1200\/JCO.20.00948","article-title":"Venetoclax Plus Rituximab in Relapsed Chronic Lymphocytic Leukemia: 4-Year Results and Evaluation of Impact of Genomic Complexity and Gene Mutations From the MURANO Phase III Study","volume":"38","author":"Kater","year":"2020","journal-title":"J. Clin. Oncol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1056\/NEJMoa2012971","article-title":"Azacitidine and Venetoclax in Previously Untreated Acute Myeloid Leukemia","volume":"383","author":"Dinardo","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2137","DOI":"10.1182\/blood.2020004856","article-title":"Venetoclax plus LDAC for newly diagnosed AML ineligible for intensive chemotherapy: A phase 3 randomized placebo-controlled trial","volume":"135","author":"Wei","year":"2020","journal-title":"Blood"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Oyogoa, E., Traer, E., Tyner, J., and Lachowiez, C. (2023). Building on Foundations: Venetoclax-Based Combinations in the Treatment of Acute Myeloid Leukemia. Cancers, 15.","DOI":"10.3390\/cancers15143589"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Kawakatsu, R., Tadagaki, K., Yamasaki, K., and Yoshida, T. (2024). Venetoclax efficacy on acute myeloid leukemia is enhanced by the combination with butyrate. Sci. Rep., 14.","DOI":"10.1038\/s41598-024-55286-0"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Kawakatsu, R., Tadagaki, K., Yamasaki, K., Kuwahara, Y., Nakada, S., and Yoshida, T. (2024). The combination of venetoclax and quer-cetin exerts a cytotoxic effect on acute myeloid leukemia. Sci Rep., 14.","DOI":"10.1038\/s41598-024-78221-9"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"36734","DOI":"10.1074\/jbc.M101287200","article-title":"Histone deacetylase is a direct target of valproic acid, a potent anticonvulsant, mood stabilizer, and teratogen","volume":"276","author":"Phiel","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"6969","DOI":"10.1093\/emboj\/20.24.6969","article-title":"Valproic acid defines a novel class of HDAC in-hibitors inducing differentiation of transformed cells","volume":"20","author":"Minucci","year":"2001","journal-title":"EMBO J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"492","DOI":"10.1002\/med.10017","article-title":"Anti-tumor mechanisms of valproate: A novel role for an old drug","volume":"22","author":"Blaheta","year":"2002","journal-title":"Med. Res. Rev."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1016\/j.leukres.2004.11.022","article-title":"Antileukemia activity of the combination of 5-aza-2\u2032-deoxycytidine with valproic acid","volume":"29","author":"Yang","year":"2005","journal-title":"Leuk. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1335","DOI":"10.1016\/j.leukres.2005.04.018","article-title":"Increase in Ara-C cytotoxicity in the presence of valproate, a histone deacetylase inhibitor, is associated with the concurrent expression of cyclin D1 and p27(Kip 1) in acute myeloblastic leukemia cells","volume":"29","author":"Siitonen","year":"2005","journal-title":"Leuk. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"10309","DOI":"10.1073\/pnas.0504248102","article-title":"Valproate activates bovine leukemia virus gene ex-pression, triggers apoptosis, and induces leukemia\/lymphoma regression in vivo","volume":"102","author":"Achachi","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","first-page":"2","article-title":"Histone deacetylase-targeted treatment restores retinoic acid signaling and differentiation in acute myeloid leukemia","volume":"61","author":"Ferrara","year":"2001","journal-title":"Cancer Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1266","DOI":"10.1182\/blood-2003-12-4333","article-title":"Treatment of myelodysplastic syndromes with valproic acid alone or in combination with all-trans retinoic acid","volume":"104","author":"Kuendgen","year":"2004","journal-title":"Blood"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1002\/cncr.21552","article-title":"The histone deacetylase (HDAC) inhibitor valproic acid as monotherapy or in combination with all-trans retinoic acid in patients with acute myeloid leukemia","volume":"106","author":"Kuendgen","year":"2006","journal-title":"Cancer"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"368","DOI":"10.1002\/ajh.23116","article-title":"Disease-stabilizing treatment with all-trans retinoic acid and valproic acid in acute myeloid leukemia: Serum hsp70 and hsp90 levels and serum cytokine profiles are determined by the disease, patient age, and anti-leukemic treatment","volume":"87","author":"Fredly","year":"2012","journal-title":"Am. J. Hematol."},{"key":"ref_40","first-page":"986","article-title":"Valproic acid and all-trans retinoic acid for the treatment of elderly patients with acute myeloid leukemia","volume":"90","author":"Raffoux","year":"2005","journal-title":"Haematologica"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1200\/JCO.19.01053","article-title":"Valproate and Retinoic Acid in Combination With Decitabine in Elderly Nonfit Patients With Acute Myeloid Leukemia: Results of a Multicenter, Randomized, 2 \u00d7 2, Phase II Trial","volume":"38","author":"Grishina","year":"2020","journal-title":"J. Clin. Oncol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1038\/s41375-018-0293-8","article-title":"Combination treatment of acute myeloid leukemia cells with DNMT and HDAC inhibitors: Predominant synergistic gene downregulation associated with gene body demethyla-tion","volume":"33","author":"Schlosser","year":"2019","journal-title":"Leukemia"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"4027","DOI":"10.1182\/blood-2013-12-546283","article-title":"Valproic acid in combination with all-trans retinoic acid and intensive therapy for acute myeloid leukemia in older patients","volume":"123","author":"Tassara","year":"2014","journal-title":"Blood"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"556","DOI":"10.1002\/cncr.29085","article-title":"Results of phase 2 randomized study of low-dose decitabine with or without valproic acid in patients with myelodysplastic syndrome and acute myelogenous leukemia","volume":"121","author":"Issa","year":"2015","journal-title":"Cancer"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Wen, J., Chen, Y., Yang, J., Dai, C., Yu, S., Zhong, W., Liu, L., He, C., Zhang, W., and Yang, T. (2023). Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia. J. Immunother. Cancer, 11.","DOI":"10.1136\/jitc-2023-006857"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1083","DOI":"10.1172\/JCI0215623","article-title":"Neutrophil-independent mechanisms of caspase-1- and IL-18-mediated ischemic acute tubular necrosis in mice","volume":"110","author":"Melnikov","year":"2002","journal-title":"J. Clin. Investig."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1023\/A:1024116916932","article-title":"Q-VD-OPh, a broad spectrum caspase inhibitor with potent antiapoptotic properties","volume":"8","author":"Caserta","year":"2003","journal-title":"Apoptosis"},{"key":"ref_48","first-page":"300","article-title":"Caspase activation in transgenic mice with Alzheimer-like pathology: Results from a pilot study utilizing the caspase inhibitor, Q-VD-OPh","volume":"2","author":"Rohn","year":"2009","journal-title":"Int. J. Clin. Exp. Med."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"791","DOI":"10.1182\/blood.2019003988","article-title":"Molecular patterns of response treatment failure after frontline venetoclax combinations in older patients with AML","volume":"135","author":"DiNardo","year":"2020","journal-title":"Blood"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1158\/2159-8290.CD-19-0710","article-title":"Monocytic Subclones Confer Resistance to Ve-netoclax-Based Therapy in Patients with Acute Myeloid Leukemia","volume":"10","author":"Pei","year":"2020","journal-title":"Cancer Discov."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1630","DOI":"10.1016\/S1470-2045(20)30525-8","article-title":"Venetoclax or placebo in combination with bortezomib and dexamethasone in patients with relapsed or refractory multiple myeloma (BELLINI): A randomised, double-blind, multicentre, phase 3 trial","volume":"21","author":"Kumar","year":"2020","journal-title":"Lancet Oncol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/S0378-4274(02)00422-8","article-title":"TRAIL-induced signalling and apoptosis","volume":"139","author":"MacFarlane","year":"2003","journal-title":"Toxicol. Lett."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1172\/JCI6926","article-title":"Safety and antitumor activity of recombinant soluble Apo2 ligand","volume":"104","author":"Ashkenazi","year":"1999","journal-title":"J. Clin. Investig."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1038\/5517","article-title":"Tumoricidal activity of tumor necrosis factor\u2013related apoptosis\u2013inducing ligand in vivo","volume":"5","author":"Walczak","year":"1999","journal-title":"Nat. Med."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Yoshida, T., Yamasaki, K., Tadagaki, K., Kuwahara, Y., Matsumoto, A., Sofovic, A.E., Kondo, N., Sakai, T., and Okuda, T. (2022). Tumor necrosis factor related apopto-sis inducing ligand is a novel transcriptional target of runt related transcription factor 1. Int. J. Oncol., 60.","DOI":"10.3892\/ijo.2021.5296"}],"container-title":["Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-9721\/13\/1\/10\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:25:15Z","timestamp":1759919115000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-9721\/13\/1\/10"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,8]]},"references-count":55,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,1]]}},"alternative-id":["diseases13010010"],"URL":"https:\/\/doi.org\/10.3390\/diseases13010010","relation":{},"ISSN":["2079-9721"],"issn-type":[{"value":"2079-9721","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,8]]}}}