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Valproic acid (VPA) is an antiepileptic drug that acts partially by inhibiting histone deacetylases (HDACs) and could be repurposed as an epigenetic anticancer therapy. Here, we show that VPA reduced medulloblastoma (MB) cell viability and led to cell cycle arrest. These effects were accompanied by enhanced H3K9 histone acetylation (H3K9ac) and decreased expression of the MYC oncogene. VPA impaired the expansion of MB neurospheres enriched in stemness markers and reduced MYC while increasing TP53 expression in these neurospheres. In addition, VPA induced morphological changes consistent with neuronal differentiation and the increased expression of differentiation marker genes TUBB3 and ENO2. The expression of stemness genes SOX2, NES, and PRTG was differentially affected by VPA in MB cells with different TP53 status. VPA increased H3K9 occupancy of the promoter region of TP53. Among the genes regulated by VPA, the stemness regulators MYC and NES showed an association with patient survival in specific MB subgroups. Our results indicate that VPA may exert antitumor effects in MB by influencing histone acetylation, which may result in the modulation of stemness, neuronal differentiation, and the expression of genes associated with patient prognosis in specific molecular subgroups. Importantly, the actions of VPA in MB cells and neurospheres include a reduction in the expression of MYC and an increase in TP53.<\/jats:p>","DOI":"10.3390\/cells14020072","type":"journal-article","created":{"date-parts":[[2025,1,7]],"date-time":"2025-01-07T10:45:55Z","timestamp":1736246755000},"page":"72","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Modulation of Stemness and Differentiation Regulators by Valproic Acid in Medulloblastoma Neurospheres"],"prefix":"10.3390","volume":"14","author":[{"given":"Nat\u00e1lia Hogetop","family":"Freire","sequence":"first","affiliation":[{"name":"Children\u2019s Cancer Institute (ICI), Porto Alegre 90620-110, RS, Brazil"},{"name":"Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"},{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alice Laschuk","family":"Herlinger","sequence":"additional","affiliation":[{"name":"Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"},{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9382-5460","authenticated-orcid":false,"given":"Julia","family":"Vanini","sequence":"additional","affiliation":[{"name":"Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"},{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matheus","family":"Dalmolin","sequence":"additional","affiliation":[{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"},{"name":"InovAI Lab, nPITI\/IMD, Federal University of Rio Grande do Norte, Natal 59078-970, RN, Brazil"},{"name":"Bioinformatics Multidisciplinary Environment (BioME), Federal University of Rio Grande do Norte, Natal 59078-970, RN, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7536-2506","authenticated-orcid":false,"given":"Marcelo A. C.","family":"Fernandes","sequence":"additional","affiliation":[{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"},{"name":"InovAI Lab, nPITI\/IMD, Federal University of Rio Grande do Norte, Natal 59078-970, RN, Brazil"},{"name":"Bioinformatics Multidisciplinary Environment (BioME), Federal University of Rio Grande do Norte, Natal 59078-970, RN, Brazil"},{"name":"Department of Computer Engineering and Automation, Federal University of Rio Grande do Norte, Natal 59078-970, RN, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carolina","family":"N\u00f6r","sequence":"additional","affiliation":[{"name":"The Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada"},{"name":"Developmental and Stem Cell Biology Program, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6557-895X","authenticated-orcid":false,"given":"Vijay","family":"Ramaswamy","sequence":"additional","affiliation":[{"name":"The Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada"},{"name":"Developmental and Stem Cell Biology Program, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada"},{"name":"Division of Haematology\/Oncology, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada"},{"name":"Department of Medical Biophysics, University of Toronto, Toronto, ON M5G 1L7, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Caroline Brunetto","family":"de Farias","sequence":"additional","affiliation":[{"name":"Children\u2019s Cancer Institute (ICI), Porto Alegre 90620-110, RS, Brazil"},{"name":"Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"},{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7958-1279","authenticated-orcid":false,"given":"Andr\u00e9 Tesainer","family":"Brunetto","sequence":"additional","affiliation":[{"name":"Children\u2019s Cancer Institute (ICI), Porto Alegre 90620-110, RS, Brazil"},{"name":"Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"},{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0668-6894","authenticated-orcid":false,"given":"Algemir Lunardi","family":"Brunetto","sequence":"additional","affiliation":[{"name":"Children\u2019s Cancer Institute (ICI), Porto Alegre 90620-110, RS, Brazil"},{"name":"Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"},{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lauro Jos\u00e9","family":"Gregianin","sequence":"additional","affiliation":[{"name":"Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"},{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"},{"name":"Department of Pediatrics, School of Medicine, Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"},{"name":"Pediatric Oncology Service, Clinical Hospital, Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mariane da Cunha","family":"Jaeger","sequence":"additional","affiliation":[{"name":"Children\u2019s Cancer Institute (ICI), Porto Alegre 90620-110, RS, Brazil"},{"name":"Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"},{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael D.","family":"Taylor","sequence":"additional","affiliation":[{"name":"The Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada"},{"name":"Developmental and Stem Cell Biology Program, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada"},{"name":"Department of Medical Biophysics, University of Toronto, Toronto, ON M5G 1L7, Canada"},{"name":"Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada"},{"name":"Department of Surgery, University of Toronto, Toronto, ON M5T 1P5, Canada"},{"name":"Texas Children\u2019s Cancer and Hematology Center, Houston, TX 77030, USA"},{"name":"Department of Pediatrics\u2014Hematology\/Oncology, Baylor College of Medicine, Houston, TX 77030, USA"},{"name":"Department of Neurosurgery, Baylor College of Medicine, Houston, TX 77030, USA"},{"name":"Department of Neurosurgery, Texas Children\u2019s Hospital, Houston, TX 77030, USA"},{"name":"Dan L Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX 77030, USA"},{"name":"Hematology-Oncology Section, Texas Children\u2019s Cancer Center, Houston, TX 77030, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6016-2261","authenticated-orcid":false,"given":"Rafael","family":"Roesler","sequence":"additional","affiliation":[{"name":"Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"},{"name":"National Science and Technology Institute for Children\u2019s Cancer Biology and Pediatric Oncology-INCT BioOncoPed, Porto Alegre 90035-003, RS, Brazil"},{"name":"Department of Pharmacology, Institute for Basic Health Sciences, Federal University of Rio Grande do Sul, Porto Alegre 90035-003, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.ccr.2008.07.005","article-title":"Acquisition of granule neuron precursor identity is a critical determinant of progenitor cell competence to form Shh-induced medulloblastoma","volume":"14","author":"Heine","year":"2008","journal-title":"Cancer Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.ccr.2011.12.021","article-title":"An animal model of MYC-driven medulloblastoma","volume":"21","author":"Pei","year":"2012","journal-title":"Cancer Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1038\/s41572-019-0063-6","article-title":"Medulloblastoma","volume":"5","author":"Northcott","year":"2019","journal-title":"Nat. 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