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Senbabaoglu Aksu, F.

Publications and source records attributed to Senbabaoglu Aksu, F..

3 recordsLinked to original sources

Targeting CBP/p300 Overcomes Acquired Vincristine Resistance in Medulloblastoma

BackgroundMedulloblastoma is the most common malignant pediatric brain tumor. Although advances in conventional therapies have improved survival over the years, acquired drug resistance remains a major barrier to durable cure. As dysregulation of epigenetic mechanisms is increasingly recognized as a driver of medulloblastoma pathogenesis and therapeutic adaptation, targeting epigenetic vulnerabilities represents a promising strategy to overcome treatment resistance. MethodsWe generated vincristine-resistant medulloblastoma cell line models and performed chemical screening to identify therapeutically targetable vulnerabilities. Candidate hits were validated using transcriptomic analyses, chromatin immunoprecipitation, and CRISPR-mediated genetic ablation to define the molecular mechanisms underlying drug sensitivity. ResultsChemical screening identified multiple active epigenetic compound classes capable of resensitizing vincristine-resistant medulloblastoma cells, including histone methyltransferase inhibitors, histone deacetylase inhibitors, and bromodomain inhibitors. Among these, the CBP/p300 bromodomain inhibitor SGC-CBP30 emerged as the most potent sensitizer to vincristine. Transcriptomic profiling revealed that, while ABCB1 was among the most highly upregulated genes in resistant cells, SGC-CBP30 treatment selectively downregulated ABCC3 and ABCA4, an effect not observed in parental cells. Mechanistically, chromatin immunoprecipitation demonstrated enrichment of p300 and H3K27ac at the ABCC3 and ABCA4 promoters in resistant cells, which was markedly reduced following SGC-CBP30 treatment. Consistent with these findings, genetic ablation of CREBBP or EP300 phenocopied the effects of pharmacological inhibition. Analysis of patient datasets further demonstrated elevated CREBBP, EP300, and ABCC3 expression in SHH MB, with positive correlations between ABCC3 and both CREBBP and EP300, supporting the clinical relevance of this regulatory axis. ConclusionsTogether, our findings demonstrate that CBP/p300 activity contributes to acquired vincristine-resistance in medulloblastoma. Targeting this axis represents a promising strategy to overcome drug resistance and enhance the efficacy of vincristine-based chemotherapy particularly in the context of relapsed or refractory disease. PLAIN ENGLISH SUMMARYMedulloblastoma is the most common cancerous brain tumor in children. Although many children respond well to the treatment, some tumors become resistant to chemotherapy, making them much harder to treat. Understanding why this resistance develops could lead to better treatment options for children whose cancer returns or no longer respond to therapy. In this study, we created laboratory models of medulloblastoma that had become resistant to the chemotherapy drug vincristine. We then tested a collection of drugs to identify compounds, which would restore the cancer cells sensitivity to treatment. We have discovered that several drugs were effective, with one compound, called SGC-CBP30, showing particularly strong activity. We investigated how SGC-CBP30 works and found that it decreases the activity of genes that are linked to chemotherapy resistance. Using multiple complementary experimental approaches, we confirmed that this gene-regulating pathway plays an important role in helping medulloblastoma cells survive treatment. Our findings suggest that targeting this pathway could restore the effectiveness of chemotherapy in drug-resistant tumors. Although further research is needed before this approach can be used in patients, these results provide a promising foundation for developing new treatments for children with relapsed or treatment-resistant medulloblastoma.

cancer biology↗

Chemical screens identify HDAC6 as an epigenetic vulnerability in acquired Temozolomide-resistant models of glioblastoma

Glioblastoma (GBM) is an aggressive primary brain tumor associated with a median survival of approximately 15 months following diagnosis. Current standard-of-care treatment includes surgical resection followed by radiotherapy and chemotherapy with the DNA-alkylating agent temozolomide (TMZ). However, tumor recurrence in a therapy-resistant state remains a major driver of poor patient outcomes. To investigate the molecular mechanisms underlying TMZ resistance, we generated in vitro models of acquired resistance by exposing initially TMZ-sensitive GBM cells to escalating doses of TMZ. Transcriptomic and chromatin accessibility profiling revealed extensive remodeling of DNA damage response (DDR) and DNA repair pathways that favored protection against TMZ-induced genotoxic stress. Although upregulation of O-6-methylguanine-DNA methyltransferase (MGMT) emerged as a dominant determinant of resistance in our models, the data suggested that additional adaptive resistance mechanisms contribute to the resistant phenotype. To identify targetable epigenetic dependencies associated with TMZ resistance, we performed a chemical screen using an epigenetic probe library. This screen identified multiple histone deacetylase (HDAC) inhibitors that selectively impaired the viability of TMZ-resistant cells, either as monotherapy or in combination with TMZ. Among these, HDAC6-selective inhibitors, including Ricolinostat, were particularly effective at inducing cell death in TMZ-resistant GBM models. Mechanistically, HDAC6 inhibition reduced the expression of key DDR-associated genes, while MGMT responses varied depending on cellular context and treatment duration. Furthermore, pharmacological inhibition and loss-of-function studies demonstrated that targeting HDAC6 could restore TMZ sensitivity by altering the balance of DNA repair pathway activity, potentially acting as a compensatory mechanism for MGMT-mediated resistance. Collectively, our findings identify HDAC6 as an epigenetic vulnerability in acquired TMZ-resistant GBM and support the therapeutic potential of HDAC6 inhibition as a strategy to overcome TMZ resistance in glioblastoma patients.

cancer biology↗

Functional Genomic Screens Reveal RBBP4 as a Key Regulator of Cell Cycle Progression in TMZ-Resistant Glioblastoma

Temozolomide (TMZ) remains the standard of care for glioblastoma; however, its efficacy is frequently influenced by epigenetic mechanisms, notably the methylation status of the O6-methylguanine-DNA methyltransferase (MGMT) promoter. While MGMT promoter hypermethylation is associated with enhanced responsiveness to TMZ, additional epigenetic determinants of TMZ resistance remain largely undefined. In this study, we established TMZ-resistant glioblastoma cell lines that consistently maintained their resistant phenotype both in vitro and in vivo. Transcriptomic analyses revealed a marked upregulation of MGMT expression in these models. To systematically investigate the epigenetic regulators governing TMZ resistance and cell survival, we conducted CRISPR/Cas9-based functional genomic screens using our focused Epigenetic Knock-Out Library (EPIKOL), which targets 800 chromatin regulators alongside selected positive and negative controls. These unbiased screens validated MGMT as a primary mediator of TMZ resistance, confirming the robustness of our approach. Moreover, dropout screens across multiple resistant cell line models identified Retinoblastoma Binding Protein 4 (RBBP4) as a critical vulnerability. Notably, RBBP4 knockout significantly impaired cell proliferation without affecting MGMT expression, suggesting a distinct mechanism supporting the survival of TMZ-resistant glioblastoma cells. Subsequent transcriptomic profiling following RBBP4 loss demonstrated significant downregulation of cell cycle pathways, particularly the G2/M checkpoint. Live-cell imaging and immunofluorescence analyses further revealed increased cell size and multinucleation in RBBP4-deficient cells, indicative of disrupted mitotic progression. Collectively, our results identify RBBP4 as a key regulator of cell cycle progression and survival in TMZ-resistant glioblastoma and highlight its potential as a novel epigenetic target for therapeutic intervention in recurrent disease.

cancer biology↗