Tumor Treating Fields (TTFields) are approved for glioblastoma (GBM) treatment, but predictive biomarkers remain unclear. This study evaluates TTFields effectiveness in primary GBM cell cultures and explores miRNA biomarkers in tumor tissue, plasma, and primary cell cultures. TTFields were applied to 21 primary GBM cell cultures for 72 h. Cell viability was assessed pre- and post-treatment, with parallel evaluations in control cultures. Expression levels of miRNAs-21, -26a, -34a, -181c, -181d, and -485-5p were analyzed in tumor tissue, plasma, and untreated/treatment-exposed cultures. Correlation analyses examined TTFields response and miRNA expression. Response rates varied, with a mean cell viability reduction of 48.53%. Expression of miRNA-26a in tumor tissue (p = 0.041, r = 0.502) and miRNAs-21, -26a, and -181c in untreated control cultures (p < 0.05) correlated with increased TTFields effectiveness. Linear correlations were observed for miRNAs-26a and -181c in untreated control cultures ([95% CI: 0.001938-0.01725, p = 0.016; 95% CI: 0.0000003935-0.0001641, p = 0.049). Individual GBM cell cultures respond differently to TTFields. Overexpression of miRNA-26a in native tumor tissue and overexpression of miRNAs-21, -26a and -181c in untreated control cell cultures were positively correlated with increased effectiveness of TTFields treatment.

Mass effect, characterized by the compression and deformation of neural tissue from space-occupying lesions, can lead to debilitating neurological symptoms and poses a significant clinical challenge. In the primary brain tumor glioblastoma (GBM), we have shown previously that compressive solid stress originating from the growing tumor reduces cerebral blood flow, leading to neuronal loss, increased functional impairment, and poor clinical outcomes. However, the direct effects of compression on neurons and the underlying biophysical mechanisms are poorly understood. Here, using multiscale compression systems and physiologically relevant in vitro and in vivo models, we find that chronic mechanical compression induces neuronal apoptosis and loss of synaptic puncta, leading to disrupted neural network activity, as assessed by calcium imaging. This is accompanied by increased HIF-1 signaling and upregulation of downstream stress-adaptive genes in neurons. We further show that chronic compression triggers AP-1-driven gene expression in glial cells, promoting a neuroinflammatory response. Together, these findings reveal that solid stress directly contributes to neuronal dysfunction and inflammation caused by GBM by activating distinct pathways that can be targeted in future studies for neuroprotection.

Significant heterogeneity in the immune microenvironment of Glioblastoma, IDH-wildtype (GBM), has been reported, necessitating a standardized approach to evaluate immune infiltration in the context of emerging immunotherapies. To address this, we developed and validated a standardized immunohistochemistry-based scoring system for quantifying immune cell infiltration in formalin-fixed, paraffin-embedded (FFPE) tissue. Paired primary and recurrent GBM specimens from 20 adult patients were labeled for CD3, CD8, CD45, CD68, and PD-1, and scored across six anatomical regions, including intra- and peritumoral, meningeal, and normal brain areas. The scoring system demonstrated excellent interrater (ICC = 0.932) and intrarater (ICC = 0.953) reliability. CD68+ and CD45+ cells were most numerous across all regions. CD3+ and CD8+ cells concentrated more in the perivascular area rather than within the parenchyma. No significant differences in immune infiltration were observed between primary and recurrent GBM. Cox proportional-hazards analysis showed worse survival with higher CD8+ and CD45+ infiltration in primary GBM, and higher CD45+ and CD68+ infiltration in recurrent GBM. In conclusion, we propose a feasible, cost-efficient, and robust method to assess immune infiltration on FFPE material, enabling standardized comparison of inflammation, with applications for ongoing clinical trials.

ABSTRACTGlioblastoma (GBM) is a devastating brain tumor with limited treatment success, partly because in vitro models poorly mimic in vivo complexity. This study introduces a high‐throughput 3D culture platform utilizing modular starPEG–glycosaminoglycan (GAG) hydrogels that enable independent control of extracellular matrix (ECM) cues: stiffness, cytokine affinity, matrix metalloproteinase‐responsive remodeling, and cell adhesiveness via integrin‐binding RGD peptides. This platform supports encapsulation of patient‐derived GBM cells, recreates physiologically relevant tumor microenvironments in 384‐well plates, and enables automated drug testing on primary cells. Transcriptomic analyses show that 3D cultures recapitulate primary and recurrent GBM programs‐ including hypoxia‐, immune‐, and ECM‐regulatory pathways driving growth, invasion, and resistance, without externally imposed hypoxia. The platform's versatility extends to drug screening, where single and combinatorial treatments produce reproducible cytoskeletal and transcriptomic responses. Notably, the system revealed dose‐dependent reductions in invasive filaments and spheroid architecture with 5‐fluorouracil/uridine and carmustine, demonstrating its potential for optimizing combinatorial therapies. This 3D model surpasses 2D cultures, capturing tumor‐specific molecular programs and offering a robust tool for translational research. Despite lacking vascular or immune components, its tunability, scalability, and clinical relevance make it a strong basis for advanced co‐cultures. By delivering reliable, individualized therapeutic data within a short timeframe, this model holds transformative potential for personalized GBM treatment.

Malignant diffuse gliomas (MG) of the brain (WHO grade 3-4) are highly aggressive primary tumors of central nervous system (CNS), spreading rapidly by infiltrating healthy brain tissue. In the majority of cases, tumor relapse occurs. The prognostic significance of pre-surgery factors, such as inflammatory markers, particularly, the peripheral blood counts in patients with MG is discussed and remains controversial. The aim of this study was to assess the relationship between the blood cell ratios and overall survival (OS) and relapse-free survival (RFS) in MG patients. The data on 59 MG patients were analyzed: 41 cases of primary (newly diagnosed) MG (astrocytoma (A-III, WHO grade 3, n = 8) and glioblastoma (GB, WHO grade 4, n = 33)) and 18 cases of recurrent MG (recurrent A-III (WHO grade 3, n = 7) and recurrent GB (WHO grade 4, n = 11)). Blood cell counts (peripheral blood leukocytes (PBL), platelets (Pt), neutrophils (Neu), lymphocytes (Ly), monocytes (Mo)) and NLR (Neu/Ly ratio), PLR (Pt/Ly ratio), MLR (Mo/Ly ratio), and systemic immune-inflammation index (SII)) in the preoperative period (prior to re-resection in cases of recurrent MG) were evaluated. The Kaplan - Meier and Cox regression analyses of OS/RFS were performed. The potential association between the blood counts and ratios PLR (≤146 vs. >146), NLR (≤4 vs. >4), MLR (≤0.27 vs. >0.27), SII (≤906 vs. >906), as well as sex (female vs. male) and age (≤60 vs. >61) with OS and RFS were analyzed. PBL and Neu counts, as well as NLR and SII indices, in patients with primary and recurrent GB in the pre-operative period significantly exceeded the reference values (p < 0.02). PBL, Neu, and SII significantly correlated with tumor grade. In patients with primary A-III and GB, longer OS tended to be associated with high PLR, NLR, MLR, and SII values, while in patients with recurrent GB, longer OS tended to be associated with low values of these ratios. Patients with recurrent A-III and GB showed a significant association between low pre-surgery NLR, SII and better RFS while patients with recurrent GB - significant association between low pre-surgery MLR and better RFS. Significant association between OS and sex of patients with both primary and recurrent GB was shown. The results obtained suggest the possible prognostic significance of PLR, NLR, MLR, and SII values in the treatment outcomes of MG patients.

Background/Objectives: The ineffectiveness of current treatments for glioblastoma underscores the urgent need for effective alternatives. This study aimed to investigate the effectiveness of sodium dichloroacetate (NaDCA) and a sodium valproate NaDCA combination (NaVPA–NaDCA) on formed patients’ primary cell tumors on the chick embryo chorioallantoic membrane (CAM). Methods: Glioblastoma tissue samples were obtained from three patients during tumor surgery. WHO grade IV, IDH wild-type, and a strong positive cytoplasmic GFAP reaction in tumor cells characterized the investigated glioblastoma cases. The tumor cells GBM2-2F, GBM2-3F, and GBM-4M from the patients were examined. Histological examination of tumor invasion into CAM, angiogenesis, and immunohistochemical expression of GFAP-, PCNA-, p53-, EZH2- and vimentin-positive cells were examined. Results: No difference in GFAP expression was observed between the patient’s GBM tumor tissue and the tumor formed on CAM from the same patient’s tumor cells. There were no significant differences in invasion or in the frequency of GFAP- and p53-positive cells among the study control groups. The expression of PCNA-, EZH2-, and vimentin-positive cells in control tumors varied significantly. Treatment significantly reduced the incidence of tumor invasion in GBM2-2F and GBM2-4M and did not affect GBM2-3F tumors; treatment also significantly reduced GFAP expression in GBM2-3F and GBM2-4M and did not affect GBM2-2F tumors. The treatment with NaVPA–NaDCA significantly reduced the expression of PCNA, p53, EZH2 and vimentin in the tested tumors. Conclusions: Data demonstrated an antitumor effect of NaVPA–NaDCA in an in vivo model of a patient’s primary glioblastoma cells.

The tumor microenvironment (TME) is critically implicated in glioblastoma (GBM) recurrence, therapeutic resistance, and immune evasion. By analyzing both primary and recurrent GBM from Chinese Glioma Genome Atlas (CGGA) and clinical samples, we identified significant differences in TME. To overcome the limitations of traditional murine recurrent GBM model, we successfully established a new murine model and characterized the TME differences between primary and recurrent GBM in vivo. Based on immune chemokine profiling within the CGGA dataset, we selected PLX3397, a Colony stimulating factor 1 receptor (CSF1R) inhibitor, to target recurrent GBM in our murine model. PLX3397 treatment significantly attenuated tumor growth and remodeled TME. Collectively, our study firstly analyzed TME combine a large number of database samples and clinical samples, and made the first application of PLX3397 in a murine recurrent GBM model, thereby providing a novel therapeutic strategy and experimental foundation for recurrent GBM research.

Glioblastoma multiforme (GBM) is an aggressive brain tumor with poor prognosis. Microglia are key immune cells in the GBM tumor microenvironment (TME), yet their subtype-specific impact, particularly inflammatory microglia, remains poorly understood. We analyzed single-cell RNA sequencing data to characterize inflammatory microglia patterns in GBM. Patient subgroups were compared for clinical features, molecular profiles, and predicted immunotherapy responses. Univariate Cox regression identified 35 inflammatory microglia-related genes associated with overall survival (OS). Based on these genes, patients were stratified into two reproducible subtypes (Cluster1 and Cluster2) across three independent datasets. Cluster2 displayed higher OS and better predicted immunotherapy response. It showed lower inflammatory microglia gene expression, reduced cytokine/chemokine activity, enhanced neural pathway activity, lower stromal and immune infiltration, decreased epithelial-mesenchymal transition, higher tumor purity, increased genomic instability-including IDH1, ATRX, and TP53 mutations-and lower MGMT methylation. Cluster1 exhibited the opposite pattern, with higher inflammatory gene expression, an immunosuppressive TME, and elevated MGMT methylation. Inflammatory microglia-based stratification reveals key biological heterogeneity in GBM and may guide prognosis and therapeutic decision-making.

FDA-approved oncolytic herpes simplex virus-1 (oHSV) therapy has emerged as an effective viro-immunotherapy for solid tumors. However, tumor- and tumor microenvironment (TME)-associated adaptations following viral treatment, such as feedback immune suppression, neoangiogenesis, and enhanced tumor aggressiveness, often hinder complete tumor eradication. Gaining a deeper understanding of the molecular mechanisms that limit the therapeutic efficacy of oHSV will be crucial to enhancing its clinical impact. We recently discovered that oHSV induces Insulin-like growth factor 2 (IGF2) secretion, shaping an immunosuppressive TME. Similarly, radiotherapy (RTx) activates the IGF1/IGF1R and YAP1 signaling pathways, further promoting therapeutic resistance. In this study, we investigated how oHSV-induced IGF1R/YAP1 signaling influences feedback pro-survival and proliferative pathways in tumor cells and evaluated the therapeutic potential of combining IGF1R blockade with oHSV and RTx.We first demonstrated that oHSV activates IGF1R signaling in vitro and in vivo, promoting tumor proliferation. While IGF1R-targeted monotherapies showed limited cytotoxic effects, combining IGF1R inhibitors with oHSV led to a significant, albeit modest, increase in cytotoxicity across tested in vitro breast cancer (BC) and primary glioblastoma (GBM) cells and in vivo xenograft models. Furthermore, we observed that co-treatment with oHSV and RTx robustly activated both IGF1R and YAP1 in resistant cells, revealing the IGF1R/YAP1 axis as a key mediator of resistance to dual oHSV and RTx therapy. Notably, the triple combination of oHSV, RTx, and IGF1R blockade yielded synergistic anti-tumor effects, abolished YAP1 expression, and significantly enhanced survival in orthotopic BC and GBM models. Collectively, these findings provide a strong rationale for the clinical evaluation of triple-combination therapy as a synergistic strategy to enhance the anti-tumor efficacy of oHSV and overcome RTx resistance in patients with BC and GBM.

Glioblastoma (GBM) is the most common and aggressive primary tumor of the central nervous system (CNS) among high-grade gliomas (HGG). Patients with GBM often have a poor prognosis. Glioblastoma cells have a remarkable ability to infiltrate the CNS and migrate long distances from the tumor core, making complete surgical resection impossible. GBM infiltration relies on electrical and molecular connections with nervous tissue. Among molecular interactions, connexin 43 (Cx43) appears to be a marker for HGG in mouse models. Cx43 is a multifunctional protein, and is found on the plasma membranes of glial cells – particularly astrocytes – in the form of gap junctions or hemichannels, ensuring cell-to-cell or cell-to-extracellular space communication, respectively. Cx43 can be expressed by transformed astrocytes, i.e., GBM cells. Its abnormal expression can alter the electrical state of the system, resulting in epileptic events. To study the dynamics of GBM spread in humans, we characterized tumor and peritumoral tissue from glioma patients. Using confocal microscopy and functional assays, we assessed the state of the neuroglial network in fresh brain sections. Electrical activity, Cx43 density, and glial reactivity were correlated with the degree of tumor infiltration. In parallel, primary human glioblastoma cells were labeled by lentiviral transduction and injected into organotypic tissue sections from the same patients. The glioblastoma cells were monitored until day seven in culture by time-lapse microscopy to follow tumor progression and their response to Gap19, a selective blocker of Cx43 hemichannels. Tumor tissue has a different architecture than peritumoral tissue, which is reflected in specific electrical and morphological profiles. In peritumoral tissue, implanted GBM cells spread into the neuroglial network, and blockade of Cx43 hemichannels results in tissue remodeling and GBM cell polarization, which may correspond to a migratory pattern. This study reveals that tumor progression is not an isolated phenomenon but it is driven by interactions with the surrounding environment in living brain tissue. In particular, Cx43 hemichannels emerge as key players in tumor spread and recurrence, revealing a more complex dynamic than expected. This finding challenges the traditional idea of GBM as an entity external to the brain, especially considering the rarity of its metastases.

Glioblastoma is a lethal brain cancer marked by functional plasticity driven by tumor cell-intrinsic mutations and their interplay with developmental programs. To investigate how canonical glioblastoma mutations promote functional plasticity, we have developed an isogenic human neural stem cell (NSC) model of glioblastoma by sequential addition of TERT promoter, TP53, and PDGFRA point mutations. TP53 loss-of-function increases TERT expression during serial mutagenesis, but only triple mutant NSCs reliably form lethal brain tumors in vivo that recapitulate glioblastoma. Tumor cell evolution triggers stress-related metabolic changes and transitions toward a neuronal progenitor network driven by transcription factor INSM1. INSM1 is highly expressed in human glioblastoma tumors and, during cortical development, in intermediate progenitor cells, which give rise to neurons. Remarkably, INSM1 knockdown in triple mutant NSCs and primary glioblastoma cells disrupts oncogenic gene expression and function and inhibits the in vivo tumorigenicity of triple mutant NSCs, highlighting the functional importance of an intermediate progenitor cell-like cell state in glioblastoma pathogenesis.

Background/Objectives: Glioblastoma (GBM) is a lethal form of primary brain tumor. There has been minimal improvement in overall GBM survival in recent years. To increase survival in patients with GBM, it is important to study novel GBM molecular antigens to form the basis of better diagnostics, prognostic measures, and therapeutic advancements. Our goal is to find more robust GBM-specific antigenic biomarkers to eventually improve GBM outcomes. Here, we initiated an investigation into cell-surface PCNA (csPCNA), a potential GBM biomarker and antigenic target. Methods: We utilized flow cytometry, imaging flow cytometry, and cell-surface Western blot to identify the expression of csPCNA on GBM cell lines (LN-229, LN-18) and a primary patient-derived tumor specimen. We then employed flow cytometry to study the associative co-expression of csPCNA with other biomarkers of GBM stem cells (CD44, CD49f) and GBM immunosuppression (PD-L1, TGFβRII). Results: We elucidated that LN-229, LN-18, and the primary GBM patient cells express csPCNA. We found that csPCNA is co-expressed with CD44, CD49f, PD-L1, and TGFβRII on the primary patient-derived GBM specimen. Conclusions: Our findings that csPCNA is expressed in GBM and is co-expressed with stem cell and immunosuppressive biomarkers indicate that csPCNA may be a potentially useful clinical-pathological biomarker for GBM stemness and immunosuppression.

Patients with brain cancers are diagnosed based on MRI in the clinical setting while molecular signatures offer potential therapeutic targets. The necessity to re-form molecular and imaging information motivated our meta-analysis to decipher the correlation between the MRI-classified tumor locations, gene expression, and protein signatures in GBM. We analyzed spatial and omics data alongside the assessment of post-translational modifications. We first utilized MRI data to classify GBM into 4 groups. We then integrated imaging groups with RNA-Seq and proteomic data to determine the association between tumor locations, gene signatures, and protein abundance. Furthermore, we scrutinized independent measurements of post-translational modifications in each group of MRI-classified GBM. The coherent layer of imaging and molecular data collectively showed the dysregulation of cell cycle, ECM organization, immune infiltration or surveillance in all GBM cases regardless of tumor locations. Several neuronal and synaptic-specific genes were differentially altered, indicative of aberrant neuroactivity in GBM. These dysregulated genes and networks provided druggable targets that led to small compounds identification, possessing cytotoxicity against primary GBM and spanning histological boundaries. Our analysis also revealed lesion-specific molecular signatures in each group of GBM, suggesting pathological features uniquely in subgroups of GBM with prognostic or therapeutic potential. Moreover, alterations in post-translational modifications would be noteworthy to explore clinical applications. Deliverables from our meta-analysis hold the potential to inform therapeutic intervention. Despite heterogeneity in GBM, our findings implicate new directions of emerging treatments that may be used as concomitants to chemo-, radio- or immunological therapies.

Preoperative differentiation between primary central nervous system lymphoma (PCNSL) and glioblastoma (GBM) is crucial for appropriate management and surgical planning. This study aims to evaluate the diagnostic performance of the AI-assisted workflow, LINNDA (lymphoma identification through neural network detection aid), in comparison to that of human raters. In total, ten clinicians independently reviewed 46 cases of GBM and PCNSL. The LINNDA workflow evaluated all 1,470 possible pairwise combinations. For each pair, whenever two clinicians disagreed, a DenseNet169 neural network was explicitly integrated as a third independent diagnostic opinion ("tie-breaker"). Integrating the AI-generated predictions improved overall accuracy to 89.9%, exceeding the expert consensus. We further established the superiority of our approach over a third human rater in another 5,108 possible combinatory scenarios. LINNDA has a negative predictive value of 97% for ruling out the diagnosis of PCNSL, providing a sound basis for clinical decision-making.

BackgroundSarcopenia refers to a decrease in skeletal muscle mass, and is associated with not only decreased functional capacity but also increased mortality. In recent years, the thickness of the temporalis muscle on head MRI has begun to be used as an alternative indicator of sarcopenia, and in this study, we investigated the significance of temporalis muscle thickness (TMT) as a prognostic predictor for patients with glioblastoma.MethodsThe subjects were 225 cases of primary glioblastoma treated at our hospital. TMT was measured from head MRI data, and the survival time (OS) was retrospectively examined. TMT was measured in slices in which both Sylvian fissures were clearly depicted, and the average value of the left and right was calculated. The association of TMT with age was also compared.ResultsThe median age was 68 years (30-87 years), and the median TMT was 8.09 mm (4.43-12.51 mm). When the patients were divided into two groups (High 115, Low 112) based on the median TMT, the median OS was 24.7 months and 11. 5 months, which was significantly longer in the High group (p < 0.0001). When the patients were further divided into three groups (High 74, Middle 76, Low 77), the OS was 25.8 months, 19.6 months, and 10.5 months, respectively, and the thicker TMT, the longer the OS was with a significant difference (p < 0.0001). Based on age, 111 cases were aged and 114 cases were young. TMT was mean 7.6mm and 8.2 mm, respectively, which was significantly thicker in the young group (p = 0.001), and the OS was also significantly longer in the young group (23.8 months and 15.8 months, respectively) (p = 0.0005), showing an association with age.ConclusionIn glioblastoma patients, TMT was a useful prognostic predictor. However, it is necessary to consider the values according to age.

PurposeGlioblastoma is a rapidly progressing malignancy, and earlier treatment initiation is linked to better outcomes. Prior studies have reported that surgery within three weeks of symptom onset is associated with improved prognosis. Regional general hospitals often lack the equipment and staffing found in university hospitals. At our facility, we rent an intraoperative navigation system as needed, and radiation therapy is provided by visiting physicians three times per week. This study aimed to evaluate whether standard glioblastoma treatment can be delivered in a regional hospital by examining time intervals from symptom onset to consultation, imaging, surgery, and the start of radiation therapy.Subjects and MethodsWe retrospectively analyzed data from patients with primary glioblastoma treated at our hospital between 2021 and 2025. Variables included age, sex, symptom onset date, consultation date, first imaging date, surgery date, radiation therapy initiation date, KPS at admission, and survival time. Key outcome measures were the intervals from symptom onset to surgery, onset to first imaging, consultation to surgery, and surgery to radiation therapy.ResultsSeven patients were included. The median age was 70 years (IQR: 55-80), and the median KPS at presentation was 80 (IQR: 60-80). Median intervals were 34 days (IQR: 32-54) from symptom onset to surgery, 7 days (IQR: 0-10) to initial consultation, 22 days (IQR: 18-24) from consultation to surgery, and 17 days (IQR: 15-20) from surgery to radiation therapy initiation.ConclusionDespite limited resources, our hospital achieved treatment intervals comparable to those of larger institutions. This suggests that, with efficient use of equipment and staffing, standard glioblastoma care can be effectively delivered in regional general hospitals.

The purpose of this study is to explore the role in MCTP2 glioblastoma (GBM) relapse and associated therapy target, especially its function on immune suppression together with drug-resistance. We performed transcriptional analysis of primary and recurrent GBM samples from the TCGA and CGGA databases to identify a key regulatory gene in relation to synaptic-related pathways using MCTP2. We used single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics analysis of expression in GBM samples to explore the population landscapes as well as tumor region-specific features at the molecular level. Functional studies via Western blotting, colony-formation assays and patient-derived organoid (PDO) models were used to assess the effect of MCTP2-depletion with/without SB52334 treatment in GBM. MCTP2 is significantly up-regulated in recurrent GBM and it correlates with poor survival. In addition, the increased expression of immune checkpoint markers PD-L1 (CD274) and CTLA-4 were significantly associated with high MCTP2 levels, indicating that MCTP2 could affect GBM patients' responses to anti-PD-1/PD-L1 or/and anti-CTLAs therapies. Overexpression of MCTP2 was also related to the increase in drug-resistance, specifically against SB52334. Combining MCTP2 KD with SB52334 significantly decreased primary cell clonogenic potential and organoid viability, suggesting a synergistic effect. MCTP2 plays a distinct role in GBM, acting as a novel facilitator of immune evasion and drug resistance with potential therapeutic implications. Our findings suggest that combining MCTP2 knockdown with SB52334 treatment could enhance the therapeutic efficacy against GBM.

BackgroundThe intra- and inter-tumoral heterogeneity of glioblastoma (GBM) represents a significant therapeutic challenge and barrier to the generation of reliable and accurate models for in vitro studies. Three-dimensional models, such as patient-derived neurospheres (PDN) and organoids (PDO), recapitulate complex cellular states. However, the direct comparison of models derived in parallel from the same primary tumor tissue has never been performed. The aim of this study was to determine the tumor cell composition of PDN and PDO models relative to matched primary GBM tissue.MethodsFour GBM surgical samples were used to establish matched PDO and PDN models, which were genomically verified using single nucleotide polymorphism array. DNA methylation, histology, and transcriptome were examined for intra-tumoral heterogeneity correlating with the matched GBM tissue. PDN lines were used to investigate the tumor cell composition response to temozolomide chemotherapy over time, maximizing their utility.ResultsWe find that both patient-derived models recapitulate the genomic, epigenomic, and tumor cell heterogeneity of the primary tissue at similar capacity. Furthermore, single-nuclei RNA sequencing revealed a subset of organoids containing small numbers of non-malignant cells from neuron and immune cell lineages. Harnessing the intra-tumoral heterogeneity of PDN models, we reveal the impact of temozolomide chemotherapy on individual cell states, altering composition of tumors over time in response to treatment.ConclusionsOur data confirms that both PDN and PDO patient-derived models recapitulate patient intra-tumoral heterogeneity providing a platform for tumor cell state refined therapeutic studies.

[This retracts the article DOI: 10.1155/2011/314962.].

Glioblastoma (GBM) remains one of the deadliest primary brain tumors, with rapid recurrence and near-universal resistance to temozolomide (TMZ) limiting long-term survival. In this study, we identify a clinically actionable mechanism of resistance driven by the LMNA–PRKDC axis, which enhances DNA repair and tumor cell survival following TMZ treatment. Using patient-derived xenograft models of recurrent GBM, we demonstrate that resistant tumors exhibit elevated LMNA expression and increased physical interaction with PRKDC, a central regulator of non-homologous end joining (NHEJ). This interaction accelerates the repair of TMZ-induced DNA lesions, contributing to therapeutic failure. Proteomic profiling and targeted immunoprecipitation revealed a distinct LMNA–PRKDC–associated DNA repair complex. Inhibition of PRKDC with the ATP-competitive inhibitor KU57788 reversed resistance, restoring TMZ sensitivity and impairing tumor growth in vivo. Single-cell RNA sequencing of primary and recurrent GBM specimens further identified LMNA–PRKDC co-expression as a hallmark of treatment-resistant, glioma stem-like cell populations. Importantly, high LMNA–PRKDC expression was associated with inferior survival outcomes in GBM patient cohorts. These results establish the LMNA–PRKDC axis as a functional driver of TMZ resistance through enhanced DNA repair capacity in stem-like tumor subpopulations. Our findings support pharmacologic inhibition of PRKDC as a rational strategy to resensitize resistant GBM to standard chemotherapy and offer a foundation for future biomarker-driven clinical trials targeting DNA repair vulnerabilities in recurrent disease.