Search Results (951)

Background/Objectives: Glioblastoma multiforme (GBM) treatment is limited by tumor hypoxia and poor specificity of therapeutic agents. To address these challenges, we developed brain-targeted liposomes co-encapsulating 5-aminolevulinic acid (5-ALA) and catalase (CAT), termed brain-targeted 5-ALA-CAT liposomes (BACL), which were surface-modified with the Angiopep-2 ligand to enhance blood–brain barrier penetration and achieve multimodal therapy combining targeted delivery and oxygen generation. Methods: BACL was prepared and characterized. Tumor targeting was verified by flow cytometry and in vivo imaging. In vitro antitumor activity was evaluated by wound-healing assay, colony formation assay, live/dead staining, MTT assay, and Western blotting. In vivo efficacy, apoptosis, and safety were assessed in a subcutaneous xenograft model. Transcriptome sequencing and qRT-PCR were employed to identify molecular mechanisms and novel targets. Results: BACL exhibited favorable physicochemical properties (size: 122.4 nm, PDI: 0.189, zeta potential: −12.3 mV) and spherical morphology as observed by TEM, with encapsulation efficiencies of 51.2% for 5-ALA and 43.8% for CAT. Compared with unmodified 5-ALA, BACL increased the cellular uptake efficiency by 1.6-fold in glioma cells while maintaining catalytic stability for sustained oxygen generation. In vitro experiments demonstrated that BACL significantly inhibited glioma cell migration, colony formation, and cell viability, and induced apoptosis. In a subcutaneous xenograft tumor model, BACL-mediated photodynamic therapy (PDT) achieved a tumor growth inhibition rate of 52%, with apoptosis induction via regulation of Bcl-2, Bax, and p53 expression, and no obvious toxicity to major organs was observed. Transcriptomic analysis combined with qRT-PCR validation revealed that BACL activates multiple antitumor signaling pathways, including targeted inhibition of IL-10 and CXCL13 to disrupt cytokine–receptor interactions, as well as coordinated regulation of S100A3 and IGSF-9 expression to suppress glioma progression. Conclusions: These multimodal actions enhanced PDT efficacy while remodeling the tumor microenvironment. Our findings position BACL as a promising therapeutic platform integrating targeted delivery, hypoxia alleviation, and immunomodulation for GBM therapy. Full article

Glioblastoma multiforme (GBM) is the most aggressive primary brain malignancy with limited treatment options and poor clinical outcomes. There is growing interest in using Zika virus as a treatment for GBM due to its selectivity in finding and killing rapidly proliferating neural cells. Several studies reproducibly show that Zika can effectively kill GBM cells. We sought to uncover the molecular mechanisms driving this cytotoxic effect by performing a meta-analysis of transcriptomic studies in which Zika virus was used to kill GBM cells. We integrated four datasets from studies on GBM and added neuroblastoma (NBM) studies as an outgroup comparator. Our analysis identified a shared molecular signature of the Zika-infected GBM cell. Interestingly, GBM cells killed by the Zika virus showed dysregulation of pathways commonly implicated in proliferation and metastasis, including TNF, NF-κB, and p53 signaling. Using a hypothesis-free design, we found several long non-coding RNAs (lncRNAs) that were consistently dysregulated in Zika-infected GBMs, many of which have previously unrecognized roles in cancer cell death. Among this group, we validated four lncRNAs for a role in Zika-mediated oncolysis. We functionally tested MELTF-AS1, TIPARP-AS1, NR2F1-AS1, and SLC9A3-AS1 in adult GBM cell lines using siRNA-mediated knockdown. Silencing of MELTF-AS1 augmented Zika-induced cell death, while knockdown of TIPARP-AS1, NR2F1-AS1, and SLC9A3-AS1 attenuated oncolysis, identifying lncRNAs whose modulation is associated with altered Zika-mediated cytotoxicity. These findings elucidate candidate mechanisms of Zika oncolysis in GBM cell lines, highlight novel lncRNA targets, and support further exploration of lncRNA modulation as a strategy to enhance oncolytic virotherapy for GBM and related malignancies. Full article

Background/Objectives: In this study, we develop and validate an interpretable machine learning (ML) model that integrates a hybrid swarm intelligence (SI)-based feature selection method with multiparametric magnetic resonance imaging (MRI)-derived RFs to estimate overall survival (OS) in glioblastoma multiforme (GBM) patients. Methods: A cohort of 276 GBM patients with open-access pre-treatment MRI data was used to perform comprehensive radiomic analysis. In the training (discovery) dataset, we employed five-fold cross-validation combined with bootstrapping to ensure robust methodological validation. Model evaluation covered the concordance index (C-index) with 95% confidence intervals (CIs). Additionally, survival stratification was performed using Kaplan–Meier curves and the log-rank test to separate patients into low- and high-risk groups for OS. The final survival model integrates patient age and ten independent RFs. Results: The model’s performance in the holdout test dataset was evaluated by a C-index of 0.71 (95% CI: 0.63–0.80), exhibiting statistically significant risk stratification (p = 3 × 10⁻⁴). Upon external validation, the model achieved a C-index of 0.67, maintaining statistical significance (p = 1 × 10⁻²). Conclusions: The research combined a traditional regularized Cox regression (Cox-LASSO) model with a new SI-based LASSO-PSO method, yielding significant stratification. To our knowledge, the present study offers one of the first studies to document the use of an interpretable ML model with an SI-based approach for successful risk stratification based on OS. Full article

Adverse childhood events (ACEs) represent a major public health burden in the United States, as exposure to ACEs increases the risk of mental health disorders, cardiovascular disease, obesity, and cancer and can greatly affect quality of life. Although the effects of ACEs on physiology are complex, significant evidence suggests that ACEs may affect the immune system later in life. As the field of immunotherapy gains prominence in cancer treatment, it becomes imperative to explore how ACEs may impact cancer immunotherapy. Brain cancer, especially glioblastoma multiforme (GBM), has a very poor prognosis and remains a challenging disease to treat. Immunotherapy approaches are being actively investigated. To address this, we aim to raise awareness of the potential connection between ACEs, brain cancer, and cancer immunotherapy. Through a review of the current biomedical literature, we synthesized current findings into a commentary on how early-life adversity may influence the immune response within the central nervous system. It is the intention of this commentary to generate hypothesis-driven research within the nascent and emerging field of neuro-psycho-immunology in neuro-oncology. Given that the relationship between ACEs and brain cancer immunotherapy remains vastly understudied, this commentary has significant potential to generate insightful discussions among clinicians and scientists to guide further clinical and basic research towards anticipating which patients stand to receive the most benefit from immunotherapy. Full article

Glioblastoma multiforme (GBM) is the most aggressive brain tumor in adults. Necrosis, and by inference hypoxia, is a pathognomonic feature of GBM tumors, where hypoxia significantly contributes to chemoresistance, leading to local treatment failure and disease progression. Although temozolomide (TMZ) is the main treatment option, 60–75% of GBM patients do not benefit from it. This study aimed to evaluate the therapeutic potential of Tetrandrine (TET) in combination with or compared to TMZ in treating GBM cells (M010b and U87) under both normoxic and hypoxic conditions. The therapeutic potential was assessed using qRT-PCR, MTT assay, combination index analysis, flow cytometry for apoptosis and cell cycle analysis, scratch assay, gelatin zymography, measurement of mitochondrial membrane potential (ΔΨm), reactive oxygen species (ROS) production, and molecular docking. Under both normoxic and hypoxic conditions, TET showed significant cytotoxicity in both cell lines compared to TMZ. A synergistic effect was observed only under normoxia at 2× IC₅₀ concentrations in M010b cells, and at 4× IC₅₀ concentrations in U87 cells. TET significantly increased the sub-G1 cell population and apoptosis compared to TMZ in both cell lines under normoxic and hypoxic conditions, while TMZ induced G2/M arrest in U87 cells under both conditions. TET significantly increased ROS production in both cell lines under normoxia. Under both conditions, ΔΨm was significantly reduced by TET in M010b cells and by TMZ in both cell lines. TET and TMZ significantly reduced pro-MMP-2 levels in M010b cells under both conditions and in U87 cells under normoxia. In conclusion, given the limited therapeutic potential of TMZ, our findings suggest that TET could be a viable alternative treatment option for GBM. Full article

Preclinical studies and data from other cancers suggest that inhibition of the Hedgehog (Hh) pathway also has antiproliferative effects on glioma cells. A key component of this pathway is the Smoothened (SMO) protein, which is expressed in high-grade gliomas. Itraconazole (ITRA), a widely used antifungal agent, inhibits SMO, the PI3K/AKT/mTOR pathway, and the VEGF/VEGFR-2 axis—both of which are critical for GBM progression and angiogenesis. This protocol describes a prospective, single-center, dose-escalation phase I study with the classical 3 + 3 design in order to determine the MTD of ITRA. The study enrolls patients with a newly histologically confirmed diagnosis of GBM, without previous treatment except surgery. They will be treated with standard RT schedule (60 Gy in 30 fractions) with concurrent TMZ 75 mg/m² and ITRA 2 × 100 mg, 200 mg, or 300 mg daily. The primary endpoint is to determine the MTD of ITRA given concurrently with RT + TMZ. Secondary endpoints include safety and tolerability of ITRA, overall survival (OS), progression-free survival (PFS), overall objective response rate, use of corticosteroids, treatment compliance, and health-related quality of life (EORTC QLQ-C30 and BN20). Participants will be monitored for one week post-treatment. All relevant statistics will be primarily descriptive. Full article

Extracellular vesicles (EVs) are attracting considerable interest due to their important role in cell signaling. However, their nanosized scale, complexity, and heterogeneity make even morphological characterization challenging. Only with the recent advances in cryogenic electron microscopy (cryo-EM), together with the increasing availability of cryo-electron microscopes, has it become possible to visualize the native structure of EVs. In this study we performed an in-depth cryo-EM analysis of EVs derived from four glioblastoma multiforme (GBM) cell lines (U87MG, U373MG, U251MG, and T98G), highlighting the morphological changes induced by temozolomide (TMZ) chemotherapeutic treatment. Size, shape, circularity, concentricity, membrane thickness, and electron density of EVs were analyzed. The key characteristic revealed by cryo-EM was that EVs can be enclosed not only by one membrane bilayer, but also by two or more bilayers (double-layered vesicles, DVs; and multilayered vesicles, MVs). Overall, TMZ treatment substantially modified both the morphology and production of EVs, decreasing the percentage of single-layered vesicles (SVs) while increasing that of DVs and MVs, as well as reducing the electron density of the EV cargo. Morphometric and morphological information can shed light on the contribution of EVs to tumor progression, metabolism, drug resistance, and immune evasion. Full article

Gliomas represent the most prevalent type of brain tumor, with their most aggressive variant, glioblastoma multiforme, associated with high mortality rates. Due to their elevated molecular heterogeneity, accurate classification of gliomas has presented significant challenges. Therefore, considerable effort has been dedicated to identifying relevant biomarkers that improve early diagnosis and unveil new areas for treatment. Advances in high-throughput sequencing technology have enabled public resources such as The Cancer Genome Atlas (TCGA) to provide large-scale data from various cancers, allowing researchers to perform more comprehensive analysis of this disease. In this study, we introduce MOHVAE-B, a comprehensive framework designed for the integration of multi-omics data and biomarker discovery using data from TCGA. MOHVAE-B employs a supervised hierarchical variational autoencoder integrated with SHAP-based interpretability to effectively integrate high-dimensional multi-omics data and extract the most influential features driving the model’s predictions. Subsequently, Bayesian Networks (BNs) are constructed to model conditional dependencies between the selected features, providing insights into their possible relations. Applied to the TCGA glioma cohorts, MOHVAE-B achieved a near-perfect AUC of $0.9993$ and successfully identified high-impact features related to glioma classification. For glioblastoma multiforme, this included six novel candidates: LINC02172, NACA2, LINC01114, HNRNPA1P48, PPIAL4G, and LINC01558. For low-grade gliomas, the model highlighted AMER2 as a promising marker. Across both cohorts, PMP2 stood out as a particularly strong candidate for a potential role in glioma pathogenesis. The constructed BNs provided an additional layer of validation, reinforcing NACA2 as a candidate of interest in glioma biology. Full article

Glioblastoma multiforme (GBM) remains the most aggressive and therapeutically intractable primary brain tumor, with many patients experiencing rapid relapse despite maximal surgical resection followed by standard chemoradiation. This persistent failure reflects the convergence of profound tumor-intrinsic genetic heterogeneity and a highly dynamic, spatially structured, and immunosuppressive tumor microenvironment (TME). Together, these forces create strong selective pressures that fuel tumor evolution, intratumoral diversity, phenotype plasticity, diffuse invasion, and robust resistance to therapy. The TME of GBM is orchestrated through a complex interplay between diverse cellular constituents, including tumor-associated macrophages, reactive astrocytes, endothelial cells, pericytes, and GBM stem cells, and non-cellular components such as extracellular matrix remodeling, hypoxia, metabolic and nutrient gradients, and spatially patterned cytokine and chemokine signaling networks. Additionally, heterogeneity in blood–brain barrier (BBB) and blood–tumor barrier (BTB) complicates drug delivery and immune surveillance, reinforcing therapeutic resistance and regional tumor adaptation. Conventional two-dimensional cell cultures and animal models fail to sufficiently capture these multiscale, patient-specific interactions, limiting their translational predictive power. In this narrative review, we synthesize recent advances in GBM organoid technologies as physiologically relevant, three-dimensional platforms that more faithfully recapitulate TME for driving tumor evolution and treatment resistance. We compare complementary organoid strategies, including patient-derived GBM organoids that preserve native cytoarchitecture, cerebral organoid co-culture systems that reconstruct tumor–brain interactions, and advanced platforms incorporating immune and vascular features such as air–liquid interface cultures, microglia-enriched systems, and BBB/BTB-integrated models. Finally, we highlight emerging innovations such as spatial transcriptomics, organoid-on-a-chip systems, live imaging coupled with lineage tracing, genome engineering, and artificial intelligence integration that collectively position GBM organoids at the forefront of precision neuro-oncology, reproducing TME, enabling dynamic mapping of tumor evolution, and accelerating patient-specific therapeutic discovery. Full article

Glioblastoma multiforme (GBM) is the most aggressive primary brain tumor, with well-documented incidence disparities across ethnic populations: highest in Europeans and lowest in East Asians and Africans. Still, the genetic basis of these differences remains poorly understood. This study assessed whether population-level differences in GBM risk allele frequencies correlate with ethnic disparities in prevalence. We analyzed 673 genome-wide significant GBM candidate loci across five ethnic superpopulations and 26 subpopulations using phased genotype data from the 1000 Genomes Project Phase 3. Population genetic structure was characterized using allele frequencies, heterozygosity, Wright’s fixation index, analysis of molecular variance (AMOVA), Nei’s genetic distances, and principal coordinate analysis. Risk allele enrichment was visualized via hypergeometric heatmaps, and polygenic risk scores were compared using Kruskal–Wallis and Dunn’s tests. Significant interpopulation differentiation was detected across all superpopulation pairs (p < 0.001). European populations had the highest polygenic risk scores, followed by South Asian and Admixed American populations, while East Asians had the lowest. Allele frequencies at key loci, including rs634537 (CDKN2B-AS1) and rs55705857 (CCDC26), differed up to tenfold. Finnish populations showed an elevated risk consistent with founder effects. Population genetic structure at GBM risk loci correlates with ethnic incidence disparities, underscoring the need for ancestry-specific approaches in risk modeling and trans-ancestry studies. Full article

This review demonstrates that the diagnostic and prognostic significance of glial fibrillary acidic protein (GFAP) is not limited to its use as a marker of astrocytic damage but should also be considered in the context of the diversity of GFAP isoforms, their heterogeneous tissue-specific expression and their pronounced association with extracellular vesicles (EVs). The data presented in this review indicate that GFAP-positive (GFAP+) EVs possess broad clinical relevance in both acute and chronic pathologies of the nervous system, including ischemic stroke, traumatic brain injury, glioblastoma, and potentially diabetic and drug-induced polyneuropathy. Particular attention is given to the critical analysis of methodological approaches for studying GFAP+ EVs, including discussion of their proposed biogenesis, mechanisms of intravesicular incorporation of cytoskeletal fragments, and the hypothetical sorption of GFAP within the vesicular protein corona. A principal conclusion of this work is that, despite the high translational potential of GFAP+ vesicles as a novel liquid biopsy platform, further implementation of this approach in clinical practice will require standardization of EV isolation protocols, harmonization of phenotyping methodologies in accordance with MISEV 2023 recommendations, and large-scale prospective studies aimed at validating the biological nature, origin, and clinical reproducibility of identified GFAP-associated vesicular subpopulations. Full article

Chemotherapy-induced metabolic reprogramming of glioblastoma multiforme (GBM) cells increases intracellular levels of reductive and energetic carriers, thereby fueling drug-relocation and retention systems and enhancing GBM drug-resistance. We have previously shown the role of this process in the adaptation of poly(morpho)nuclear “giant” cells (PGCs) in T98G populations to doxorubicin (DOX)-induced stress. Here, we addressed the role of a “resistance triad”, which coordinates metabolic T98G reprogramming with the activation of the drug-relocation and drug-retention axis, in the recovery of GBM populations from chemotherapeutic stress. A combination of proteomic analyses with metabolic and phenotypic profiling of pulse DOX-treated T98G cells revealed the significance of mitochondrial dynamics for the efficiency of the T98G “resistance triad”. DOX-induced mobilization of ATP-generating systems and ATP-dependent anabolic pathways was accompanied by the formation of DOX-negative, “mosaic” mitochondrial networks and the upregulation of mitofusin-2 (MFN2) in T98G PGCs. Transient MFN2 down-regulation correlated with the respiratory capacity of T98G cells, while impairing cell welfare in the absence and presence of DOX. However, minute fractions of PGCs, which withstood combined MFN2 down-regulation and pulse DOX treatment, retained mitochondrial networks and displayed efficient ABC transporter-/V-type channel-dependent lysosomal DOX retention. Collectively, a “triad” of mitochondrial activation, ABC transporter-dependent perinuclear redistribution and V-type channel-mediated lysosomal DOX compartmentalization determines DOX resistance of T98G cells. Whereas MFN2-dependent mitochondrial rearrangements may contribute to these processes, complementary adaptative mechanisms can compensate MFN2 dysfunction, limiting its potential as a therapeutic target. Full article

Background: High-grade glioma (HGG), formerly known as Glioblastoma multiforme, can be complicated by hematomas, either at the initial presentation or after surgical removal. Rarely, postoperative bleeding can occur extra-axially, resulting in subdural hematomas (SDH) that might require surgical evacuation. Of note, little is known about the cellular composition of those hematomas. Observations: We present the case of a patient operated on for HGG who developed postoperative recurrent SDH that required multiple surgical evacuations. Histopathological analyses of the membranes comprising the SDH revealed the presence of HGG tumoral cells. Conclusions: Based on our observation, hematomas associated with HGG, either extra or intra-axial, should be suspected of being a sign of tumoral recurrence or spread and histopathological analyses might be considered as they could lead to further adaptation of the patient’s treatment. Full article

Background: Glioblastoma multiforme (GBM) is the most aggressive primary brain tumor with limited treatment options. Tumors harboring the BRAF^V600E mutation exhibit aggressive behavior and present therapeutic challenges. Although dabrafenib/trametinib (D+T) target the BRAF/MAPK pathway and show efficacy in BRAF^V600E mutant melanoma, their effectiveness against GBM remains unclear. RES demonstrates anti-GBM activity through the inhibition of multiple signaling pathways. This study evaluated the therapeutic potential of RES either in monotherapy or in combination with D+T in GBM cells with and without the BRAF^V600E mutation. Methods: BRAF^V600E mutational status was confirmed in LN428 and U251 GBM cell lines using Sanger sequencing. Cell proliferation and viability was assessed by CCK-8, EdU assay and Calcein AM/PI staining, cell morphology by H&E staining, cell migration by Transwell assay, and apoptosis by TUNEL assay. The protein expressions of BRAF, pERK, and pSTAT3 were analyzed by Western blot, immunocytochemistry (ICC), and immunofluorescence (IF) following treatment with RES, D+T, or their combination. Statistical significance was determined using one-way ANOVA followed by Dunnett’s post hoc test with p < 0.05. Results: Sanger sequencing confirmed the presence of the BRAF^V600E mutation in the LN428 cells and its absence in the U251 cells. In the BRAF^V600E mutant LN428 cells, neither RES, D+T, nor their combination inhibited cell proliferation or migration, nor did they induce apoptosis. In contrast, RES monotherapy significantly suppressed proliferation, reduced migration, and induced apoptosis in the wild-type U251 cells, while D+T showed minimal inhibitory effects in both cell lines. Western blotting, ICC, and IF analyses revealed that RES significantly downregulated both pERK and pSTAT3 expression in the U251 cells but failed to produce similar effects in the LN428 cells. Notably, D+T treatment induced marked upregulation of pSTAT3 in both cell lines, which was effectively reversed by RES treatment in the U251 cells but not in the LN428 cells. Conclusions: RES selectively suppressed the MAPK and STAT3 signaling pathway in the BRAF wild-type U251 cells, while demonstrating no significant inhibitory effects in the BRAF mutant LN428 cells. This differential response indicates that mutational background governs MAPK/STAT3 pathway regulation, positioning RES as a promising dual-pathway inhibitor in mutation-stratified GBM therapeutics. Full article

Glioblastoma multiforme (GBM) is defined by rapid progression, high invasiveness, and a poor prognosis, with a median survival of only ≅13 months despite current treatments. Its marked genetic heterogeneity, high mutational burden, and cancer stem cell population make GBM exceptionally difficult to treat, highlighting the urgent need for more effective, multitargeted therapies. Non-coding RNAs, particularly tumor suppressor microRNAs (miRNAs), have gained attention for suppressing key oncogenic processes that drive tumorigenesis, metastasis, and drug resistance, positioning them as promising tools for targeting multiple oncogenic pathways. We recently found that FOXM1/AXL-eEF2K collaboratively drive GBM cell proliferation, survival, and invasion through the formation of a signaling hub complex. In this study, we employed miRNA prediction algorithms to identify a specific miRNA, in vitro functional assays and in vivo GBM flank model to target GBM tumorigenesis by distrupting the FOXM1/AXL-eEF2K signaling hub. Our results indicated that FOXM1, AXL, and eEF2K are overexpressed in GBM patient tumors. To target the FOXM1/AXL-eEF2K signaling hub, we identified miR-449b-5p, miR-329-3p, and miR-518c as potential co-inhibitors of FOXM1/AXL-eEF2K and suppressors of cell proliferation, migration–invasion, and spheroid formation. Furthermore, the combination of miR-449b-5p, miR-329-3p, and miR-518c treatments with temozolomide led to synergistic enhancements in cell proliferation suppression and the induction of apoptosis and ferroptosis. More importantly, in vivo miR-329-3p treatment led to remarkable suppression of GBM tumor xenografts. These findings indicate that miR-329-3p-based tumor suppressor therapy may offer a multitargeted approach for GBM treatment. Full article