Glioblastoma cell lines derived from different patients are widely used in tumor biology research and drug screening. A key feature of glioblastoma is the high level of inter- and intratumor heterogeneity that accounts for treatment resistance. Our aim was to investigate whether intratumor heterogeneity is maintained in cell models. Single-cell RNA sequencing was used to investigate the cellular composition of a tumor sample and six patient-derived glioblastoma cell lines. Three cell lines preserved the mutational profile of the original tumor, whereas three others differed from their precursors. Copy-number variation analysis showed significantly rearranged genomes in all the cell lines and in the tumor sample. The tumor had the most complex cell composition, including cancer cells and microenvironmental cells. Cell lines with a conserved genome had less diverse cellularity, and during cultivation, a relative increase in the stem-cell-derived progenitors was noticed. Cell lines with genomes different from those of the primary tumors mainly contained neural progenitor cells and microenvironmental cells. The establishment of cell lines without the driver mutations that are intrinsic to the original tumors may be related to the selection of clones or cell populations during cultivation. Thus, patient-derived glioblastoma cell lines differ substantially in their cellular profile, which should be taken into account in translational studies.

Glioblastoma multiforme (GBM) accounts for almost half of all primary malignant brain tumors in adults and has a poor prognosis. Here we demonstrated the oncolytic potential of the L-16 vaccine strain of measles virus (MV) against primary human GBM cells and characterized the genetic patterns that determine the sensitivity of primary human GBM cells to oncolytic therapy. MV replicated in all GBM cells, and seven out of eight cell lines underwent complete or partial oncolysis. RNA-Seq analysis identified about 1200 differentially expressed genes (FDR < 0.05) with at least two-fold expression level change between MV-infected and uninfected cells. Among them, the most significant upregulation was observed for interferon response, apoptosis and cytokine signaling. One out of eight GBM cell lines was defective in type I interferon production and, thus, in the post-interferon response, other cells lacked expression of different cellular defense factors. Thus, none of the cell lines displayed induction of the total gene set necessary for effective inhibition of MV replication. In the resistant cells, we detected aberrant expression of metalloproteinase genes, particularly MMP3. Thus, such genes could be considered intriguing candidates for further study of factors responsible for cell sensitivity and resistance to L-16 MV infection.

Gliomas are a heterogeneous group of brain tumors, among which the most aggressive subtype is glioblastoma, accounting for 60% of cases in adults. Available systemic treatment options are few and ineffective, so new approaches to therapies for glioblastoma are in high demand. In total, 131 patients with diffuse glioma were studied. Paired tumor–normal samples were sequenced on the Illumina platform; the panel included 812 genes associated with cancer development. Molecular profiles in clinically distinct groups were investigated. In low-grade glioma (LGG) patients (n = 18), the most common mutations were IDH1/2 (78%), ATRX (33%), TP53 (33%), PIK3CA (17%), and co-deletion 1p/19q (22%). In high-grade glioma (HGG) patients (n = 113), more frequently affected genes were CDKN2A/B (33%), TERTp (71%), PTEN (60%), TP53 (27%), and EGFR (40%). The independent predictors of better prognosis were tumor grade and IDH1/2 mutations. In IDH—wildtype glioblastoma patients, a history of other precedent cancer was associated with worse overall survival (OS), while re-operation and bevacizumab therapy increased OS. Also, among genetic alterations, TERTp mutation and PTEN deletion were markers of poor prognosis. Nine patients received molecular targeted therapy, and the results were evaluated. The search for molecular changes associated with tumor growth and progression is important for diagnosis and choice of therapy.

This study evaluates the oncolytic potential of the Moscow strain of reovirus against human metastatic melanoma and glioblastoma cells. The Moscow strain effectively infects and replicates within human melanoma cell lines and primary glioblastoma cells, while sparing non-malignant human cells. Infection leads to the selective destruction of neoplastic cells, mediated by functional viral replication. A positive correlation was identified between viral RNA accumulation and tumor cell death, with no replication observed in non-malignant cells. This study highlights the critical roles of cathepsins B, L, and S as mediators of the oncolytic process. The pharmacological inhibition of these enzymes significantly attenuated reovirus-induced cytotoxicity in melanoma and glioblastoma cells. Conversely, PKR production analysis revealed minimal activation in reovirus-infected tumor cells, suggesting that the hyperactivation of the RAS-signaling pathway and subsequent PKR inhibition do not directly contribute to the selective efficacy of reovirus. Moreover, infected tumor cells exhibited features of both apoptotic and non-apoptotic death, emphasizing the intricate mechanisms of reovirus-mediated oncolysis. These findings underscore the therapeutic promise of the Moscow strain of reovirus as a selective and potent oncolytic agent for targeting melanoma and glioblastoma cells.

Introduction. Glioblastoma is one of the most aggressive malignant brain tumors characterized by low sensitivity to drug therapy, frequent recur-rence after surgical removal of the tumor, and an extremely unfavorable prognosis. Aim. To analyze the overall survival of patients with primary glioblastoma of the brain based on the glial fibrillary acidic protein (GFAP) content of in blood serum. Material and methods. 176 patients with primary malignancy grade IV glioblastoma confirmed by morphological and molecular genetic studies (99 men and 77 women) aged 18–82 years who underwent examination and treatment at the N.N. Blokhin National Medical Research Center of Oncology in the period from 2013 to 2024 were enclosed in the study. GFAP concentration in the pre-treatment blood serum was determined by enzyme immuno-assay using Human GFAP ELISA kit (Biovendor, Czech Republic). The obtained data were processed using Statistica 10 (StatSoft) and SPSS (IBM) pro-grams. Overall survival analysis was performed using the Kaplan–Meier method with the log-rank test for comparisons. Differences were considered statistically significant at p < 0.05. Results. 72 of 176 patients were alive at the end of the study, the median follow-up period being 7.8 months, while 104 patients with died within a pe-riod of 1 to 103 months the median survival being 9.1 months. The median lifespan of all patients with glioblastoma was 14.8 months. 1-year overall survival rate comprised 56.7±4.2%, 2-year – 39.2±4.3%, 3-year – 30.0±4.1%, 4- and 5-year – 26.8±4.0%, 10-year – 15.0±4.4%. The risk of death in patients with primary glioblastoma of the brain was the greatest in the first two years of follow-up from the time of diagnosis. No associations were found with patients’ age and gender, or the side of the tumor lesion. To study the relationship between long-term treatment results and serum GFAP concentrations patients were divided into two groups: 1) patients in whom GFAP was not detected (GFAP-negative) and 2) those with detectable GFAP concentrations (GFAP-positive). Median survival time of 46 GFAP-negative glioblastoma patients was 24.8 months, while in 130 GFAP-positive patients it comprised only 13.7 months (p = 0.047). No differences in the overall survival in GFAP-positive glioblastoma group depending on the serum GFAP con-centration were found. The differences in the overall survival depending on the presence or absence of GFAP in blood serum were mostly pronounced in the female group and in patients below 50 years of age. Thus, median survival of GFAP-negative glioblastoma patients below 50 years was several fold higher than in GFAP-positive patients of this age group (70.8 and 12.4 months respectively). Conclusion. The overall survival of patients with grade IV primary glioblastoma is significantly higher in GFAP-negative than in GFAP-positive group. Hence, the presence of GFAP in blood serum can be regarded as an unfavorable prognostic factor for this disease.