Treatment Of Glioblastoma Research Articles

Sophoricoside Inhibited Glioblastoma Cell Progression Through Activated AMP-Activated Protein Kinase (AMPK).

Glioblastoma (GBM) is the most common malignant primary brain tumor, with a mean survival of less than 2 years. Unique brain structures and the microenvironment, including blood-brain barriers, put great challenges on clinical drug development. Sophoricoside (Sop), an isoflavone glycoside isolated from seeds of Sophora japonica L., is one of the active constituents of traditional Chinese medicine and found to inhibit the bioactivity of cytokines (e.g., interleukin-5) and inflammatory responses, as well as to attenuate glucose and lipid metabolism in related diseases. However, the effects of Sop on cancer progression have not been systemically investigated. In this study, we performed a comprehensive investigation of Sop's function in GBM using colony formation and Transwell assays in vitro, along with subcutaneous xenograft tumor analysis in vivo. We employed RNA sequencing and bioinformatics analysis in conjunction with Western blotting (WB) and reverse transcription-quantitative polymerase chain reaction (RT-PCR) to explore the underlying mechanism. Our results demonstrated that Sop suppressed U251 cell proliferation and metastasis in vitro and inhibited the tumorigenic behavior of U251 cells in vivo. Further investigations revealed a positive correlation between the levels of activated AMP-activated protein kinase (AMPK) and Sop treatment; notably the application of the AMPK inhibitor, compound C (CC), abolished inhibitory effects of Sop on the malignant phenotype of U251 cells. These findings suggest the potential application of Sop in GBM treatment and highlight opportunities for the development of new therapeutic strategies.

DNA Damage Repair in Glioblastoma: A Novel Approach to Combat Drug Resistance.

Due to the lack of effective therapeutic approach, glioblastoma (GBM) remains one of the most malignant brain tumour. By invitro investigations on primary GBM stem cells, we highlighted one of the underlying mechanisms of drug resistance to alkylating agents, the DNA damage responses. Here, flow cytometric analysis and viability and repopulation assays were used to assess the long-term cytotoxic effect induced by the administration of a fourth-generation platinum prodrug, the (OC-6-44)-acetatodiamminedichlorido(2-(2-propynyl)octanoato) platinum(IV) named Pt(IV)Ac-POA, in comparison to the most widely used Cisplatin. The immunofluorescence studies revealed changing pathways involved in the DNA damage response mechanisms in response to the two chemotherapies, suggesting in particular the role of Poly (ADP-Ribose) polymerases in the onset of resistance to Cisplatin-induced cytotoxicity. Thus, this research provides a proof of concept for how the use of a prodrug which allows the co-administration of Cisplatin and an Histone DeACetylase inhibitors, could suppress DNA repair mechanisms, suggesting a novel effective approach in GBM treatment.

Topology optimization design of the ‘Beam Shaping Assembly’ of an AB-BNCT facility—application to the case of glioblastoma treatment

Objective. This study aims to determine the optimal structure of the Beam Shaping Assembly (BSA) for an accelerator-based boron neutron capture therapy (BNCT) facility. The aim is to maximize the possible depth of treatment for glioblastoma while ensuring that a treatment time constraint is not exceeded.Approach. To achieve this goal, we utilize a new optimization procedure known as topology optimization. This technique can accurately identify the most optimal structure of a nuclear device, in this case a BSA, to be identified among 9 × 101206possible structures for the example given in this study. The exploration of such a vast space of configurations is inaccessible to any other method available to date.Main results. The topology optimization generated Air-AlF3-LiF-LiFPE BSA has an original structure that differs significantly from the structures previously tested by the BNCT community. This structure, which combines a ring collimator and a filter cone to mimic the effect of multi-field treatment, generates unprecedented treatment depths, with a treatable depth TD = 10.01 cm and an advantage depth AD = 12.48 cm (for 15 ppm of Boron-10 in blood, with a 3.5 tumor-to-blood Boron-10 concentration ratio), or TD = 10.30 cm and AD = 12.69 cm (for 18 ppm of Boron-10). These depths are much greater than any other design proposed to date by the community. The structure also verifies the latest proposed radiation protection constraints, which set limit values on its out-of-field leakages.Significance. The findings of this study indicate that topology optimization procedures are highly beneficial for the design of BSAs. In particular, the use of ring collimators could significantly improve the quality of BNCT treatments of brain tumors.

Abstract P015: Targeting SPP1 to enhance CAR T cell therapy in glioblastoma: Implications for integrating radiation therapy in resistant solid tumors

Abstract In the evolving landscape of glioblastoma (GBM) treatment, clinical trials exploring chimeric antigen receptor (CAR) T cell therapies have shown promise; however, their therapeutic efficacy remains constrained, particularly due to the immunosuppressive tumor microenvironment (TME). Emerging evidence highlights that the TME, especially myeloid-driven suppressive networks, plays a central role in resistance to CAR T cell therapy. Despite progress in elucidating these mechanisms, significant ambiguities endure, particularly regarding patient-specific TME heterogeneity and its impact on therapeutic outcomes. Radiation therapy remains a cornerstone in GBM management, known not only for its direct cytotoxic effects but also for its ability to modulate the TME and potentially enhance immune responses. However, the combination of radiation with CAR T cell therapy has not been fully explored, particularly in the context of targeting suppressive myeloid subsets. Recent studies suggest that radiation may induce immunogenic changes within the TME, potentially enhancing CAR T cell infiltration and activity. Our study seeks to investigate how targeting key suppressive mediators, such as SPP1-expressing macrophages, can reshape the TME, setting the stage for combinatorial approaches that leverage the immunomodulatory effects of radiation therapy In this study, we utilized single-cell RNA sequencing to analyze tumors from 41 glioma patients undergoing IL13Rα2-targeted CAR T cell therapy. Our findings revealed heightened suppressive extracellular matrix (ECM) remodeling and immunosuppressive signatures driven by SPP1-expressing myeloid cells, predominantly in non-responsive tumors. Elevated SPP1 expression and increased infiltration of SPP1+ myeloid cells were associated with poor clinical outcomes. Functional transcriptomic analyses demonstrated that SPP1-high macrophages exhibited reduced antigen presentation, impaired phagocytosis, and enhanced ECM biogenesis, coupled with elevated lipid metabolism and non-glycolytic ATP production. These metabolic adaptations were linked to a suppressive immunophenotype that hindered CAR T cell efficacy. To explore therapeutic strategies, we assessed the impact of SPP1 blockade in preclinical syngeneic glioma models. Pre-treatment with anti-SPP1 antibodies prior to CAR T cell infusion significantly reprogrammed the TME, enhancing CAR T cell efficacy and reversing resistance in previously non-responsive models. These results highlight the potential of targeting SPP1 to disrupt suppressive macrophage networks, thereby augmenting CAR T cell responses. Our findings underscore SPP1 as a promising target to overcome resistance in CAR T cell therapies for GBM. This study lays the groundwork for future integration of targeted therapies, such as SPP1 inhibition, with radiation therapy to potentially improve therapeutic outcomes in resistant gliomas, offering new avenues for combinatorial therapeutic innovation. Citation Format: Sharareh Gholamin, Heini Natri, Yuqi Zhao, Shengchao Xu, Maryam Aftabizadeh, Begonya CominAnduix, Supraja Saravanakumar, Christian Masia, Robyn Wong, Lance Peter, Mei-i Chung, Evan D. Mee, Brenda Aguilar, Davis Y. Torrejon, Anusha Kalbasi, Darya Alizadeh, Xiwei Wu, Antoni Ribas, Stephen Forman, Behnam Badie, Terence M. Williams, Nicholas E. Banovich, Christine E. Brown.Targeting SPP1 to enhance CAR T cell therapy in glioblastoma: Implications for integrating radiation therapy in resistant solid tumors.[abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Targeted Therapies in Combination with Radiotherapy; 2025 Jan 26-29; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(2_Suppl):Abstract nr P015

Abstract P005: Fibroblast Activation Protein (FAP)-targeted radioligand therapy as a promising treatment for glioblastoma

Abstract Introduction: Glioblastoma (GBM) is the most common and aggressive primary brain tumor. The 5-year survival rate is only 4% despite the current standard treatment approaches of surgery, chemotherapy, and radiation. Fibroblast Activation Protein (FAP)-targeted radioligand therapy (RLT) has emerged as a potential alternative to traditional radiation methods. FAP is a protein highly expressed in Cancer-Associated Fibroblasts (CAFs) and on GBM cells. It can be targeted by radiolabeled molecules for selective tumor irradiation, to enhance treatment precision and reduce damage to healthy tissue. Here we evaluated the relevance of FAP inhibitor 46 (FAPi-46) as a molecular probe for Radioligand Therapy (RLT) in GBM models. Methods: U87MG, a human GBM cell line shown to express human FAP and SB28, a murine GBM cell line engineered to express murine FAP were used as GBM models. NSG and C57BL6J mice were, respectively, inoculated with U87MG and SB28 cells subcutaneously. Tumor growth was evaluated by computed tomography (CT). FAP expression was assessed by 68Ga-FAPi-46 PET imaging when tumors reached around 100 mm3 prior to treatment. Mice were subsequently randomized into 4 groups: (1) vehicle, (2) 5 mg/kg temozolomide (TMZ), (3) 60 kBq 225Ac-FAPi-46, and (4) 5 mg/kg TMZ and 60 kBq 225Ac-FAPi-46. Dose of TMZ and 225Ac-FAPi-46 were increased for the SB28 study. Overall survival was tracked. Blood was collected 24h before, after, and 7 days after RLT treatment to assess blood toxicity. Tumor were resected 24h after RLT treatment to further evaluate DNA-damage. Results: Our results show that combining TMZ with 225Ac-FAPi-46 successfully delays tumor progression and extends survival in immunocompromised mice bearing subcutaneous human U87MG GBM tumors. In the SB28 GBM model neither TMZ nor a single dose of FAPi-RLT alone affected tumor growth or survival, confirming its in vivo resistance to both chemotherapy and radiation. However, by adapting the 225Ac-FAPi-46-RLT regimen, we observed a significant reduction in tumor volume and an increase in survival in immunocompetent mice. These results suggest that higher doses of 225Ac-FAPi-46-RLT can overcome resistance in this model. Conclusion: In conclusion, our study shows that 225Ac-FAPi-46-RLT could be an effective treatment for GBM, with positive results across different tumor models. To further investigate the efficacy of FAP-targeted RLT for GBM, we have initiated an evaluation of immune cell infiltration within subcutaneous SB28 tumors. Using immunohistochemistry, RNA sequencing, and flow cytometry, ongoing studies will characterize the immunogenic response induced by RLT in this GBM model. Citation Format: Pauline Jeanjean, Rachel Dove, Ines Camille Azrour, Samantha Kwock, Sarah Taylor, Sarina Smolev, Johannes Czernin, Giuseppe Carlucci, David Nathanson, Christine Mona, Elie Besserey-Offroy. Fibroblast Activation Protein (FAP)-targeted radioligand therapy as a promising treatment for glioblastoma. [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Targeted Therapies in Combination with Radiotherapy; 2025 Jan 26-29; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(2_Suppl):Abstract nr P005

Predicting brain tumour growth patterns using a novel MRI-based tumour spread map: application to radiotherapy planning.

The treatment of glioblastomas (GBM) with radiation therapy is extremely challenging due to their invasive nature and high recurrence rate within normal brain tissue. In this work, we present a new metric called the tumour spread (TS) map, which utilizes diffusion tensor imaging (DTI) to predict the probable direction of tumour cells spread along fiber tracts. We hypothesized that the TS map could serve as a predictive tool for identifying patterns of likely recurrence in patients with GBM and, therefore, be used to modify the delivery of radiation treatment to pre-emptively target regions at high risk of tumour spread. In this proof-of-concept study, we visualized the white matter fiber tract pathways within the brain using diffusion tensor tractography and developed an algorithm which mathematically calculates a relative probability index in each voxel, resulting in the generation of the TS map. Based on the information provided by the TS map, the original radiotherapy target volume was then modified to include areas with a higher probability of tumour spread and exclude other areas with a lower probability of spread. A volumetric modulated arc therapy (VMAT) treatment plan was then developed utilizing the modified target volumes and subsequently compared to that using the original target volumes. Follow-up anatomical imaging obtained 8 months post-surgery was assessed to validate our findings. A TS map was generated on a glioblastoma patient demonstrating a relative probability of tumour spread along fiber tracts throughout the brain. The modified planning target volume better covered brain regions with a higher risk of tumour spread while still demonstrating a 21% reduction in volume compared to the original planning target volume, resulting in greater preservation of normal tissue. The modified VMAT plan resulted in an average mean dose to four identified recurrences of 80% of the prescription dose, while the original VMAT plan delivered only 63% of the prescription dose as the average mean dose to the recurrences. The utilization of tractography and the generation of corresponding TS maps offer a promising approach to predicting patterns of tumour recurrence and optimizing treatment delivery. Further research is needed to validate the predictive value of the TS map on a larger cohort of patients and explore its potential in personalized treatment strategies for GBM patients.

Glycosylated Delphinidins Decrease Chemoresistance to Temozolomide by Regulating NF-κB/MGMT Signaling in Glioblastoma

Glioblastoma (GB) is a highly malignant brain tumor with a poor prognosis, with a median survival of only 14.6 months despite aggressive treatments. Resistance to chemotherapy, particularly temozolomide (TMZ), is a significant challenge. The DNA repair enzyme MGMT and glioblastoma stem cells (GSCs) often mediate this resistance. Recent studies highlight the therapeutic potential of natural compounds, particularly delphinidins, found in deep purple berries. Delphinidins are known for their ability to inhibit NF-κB signaling, a critical pathway for GB progression, chemoresistance, and MGMT expression. Our research demonstrates that glycosylated delphinidins have potential adjuvant use in the treatment of GB, offering a promising natural strategy to combat TMZ resistance. Specifically, we observed that delphinidin 3,5 di-glucoside has potent anticancer effects when used alone. Meanwhile, delphinidin 3 glucoside acted in synergy with temozolomide to decrease cell viability, highlighting its potential as an adjuvant. It also exerted a faster and more sustained inhibition of NF-κB, highlighting its potential for long-lasting therapeutic effects. These findings open new avenues for targeted therapies against glioblastoma, particularly to overcome treatment resistance.

Copper-Induced Enhancement of Glioblastoma Tumorigenicity via Cytochrome C Oxidase

Copper is an essential trace element, yet chronic copper exposure can lead to toxicity in humans, and high levels of copper have been found in the blood or tumors of patients with various forms of cancer and may affect cancer severity and response to treatment. Copper is required for the activation of cytochrome c oxidase (CcO), the mitochondrial complex that facilitates oxidative phosphorylation (OXPHOS)-mediated ATP production. We recently reported that the increased activation of CcO underlies the acquisition of treatment resistance in glioblastoma (GBM) cells. However, the potential role of copper in GBM progression or treatment resistance has not been investigated. Here, we present evidence that exposure to 20 µM copper, the maximum allowable limit for public water supplies set by the U.S. Environmental Protection Agency, promotes GBM tumor growth and reduces overall survival in vivo and increases GBM cell resistance to radiation and chemotherapy in vitro. In vitro exposure to 20 µM copper substantially increased the activity of CcO, elevated the rate and level of ATP production, and triggered a metabolic shift to an OXPHOS phenotype in GBM cells. Furthermore, copper exposure led to a substantial increase in the accumulation of glutathione and glutathione precursors in these cells. These findings establish copper as a tumor promoter in GBM and suggest that copper mediates these effects through the upregulation of CcO activity, which enhances OXPHOS metabolism and glutathione production.

The natural product micheliolide promotes the nuclear translocation of GAPDH via binding to Cys247 and induces glioblastoma cell death in combination with temozolomide.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is significantly upregulated in glioblastoma (GBM) and plays a crucial role in cell apoptosis and drug resistance. Micheliolide (MCL) is a natural product with a variety of antitumour activities, and the fumarate salt form of dimethylamino MCL (DMAMCL; commercial name ACT001) has been tested in clinical trials for recurrent GBM; this compound suppresses the proliferation of GBM cells by rewiring aerobic glycolysis. Herein, we demonstrated that MCL directly targets GAPDH through covalent binding to the cysteine 247 (Cys247) residue. Intriguingly, MCL does not affect the enzyme activity of GAPDH but facilitates the nuclear translocation of the GAPDH/Siah1 (E3 ligase) complex. Furthermore, MCL/DMAMCL can exacerbate temozolomide (TMZ)-induced DNA damage. This treatment synergistically induced GBM cell death and suppressed tumour growth in a GBM xenograft mouse model. Collectively, our results reveal that MCL triggers non-glycolysis-related functions of GAPDH and that MCL promotes GBM cell death, especially when combined with TMZ, thus providing a novel strategy for clinical GBM treatment.

Targeted therapy for glioblastoma utilizing hyaluronic acid-engineered liposomes for adriamycin delivery.

Glioblastoma (GBM) is a malignant tumor with highly heterogeneous and invasive characteristics leading to a poor prognosis. The CD44 molecule, which is highly expressed in GBM, has emerged as a highly sought-after biological marker. Therapeutic strategies targeting the cell membrane protein CD44 have emerged, demonstrating novel therapeutic potential. In this study, we constructed a nanodrug system (HA-Liposome@Dox) based on hyaluronic acid-engineered liposomes delivering adriamycin to target GBM. The system efficiently encapsulated Dox inside the liposomes through a hydrophilic-hydrophobic interaction mechanism, and the resulting HA-Liposome@Dox exhibited excellent loading efficacy, attributed to its uniform particle size distribution and negatively charged surface. Further evaluation revealed that HA-Liposome@Dox possessed excellent stability and safety and could promote the effective uptake of drug particles by CD44-overexpressing tumor cells, thus exerting a more potent cell-killing effect. Notably, in the treatment of GBM, HA-Liposome@Dox demonstrated significantly greater tumor growth inhibition compared to free Dox and prolonged the survival of tumor-bearing mice. Taken together, the present study not only verified the feasibility of HA-Liposome@Dox as an effective therapeutic tool against GBM and other CD44-positively expressing tumors, but also opened a promising new avenue for the clinical treatment of this type of refractory malignancies.&#xD.

Severe Temozolomide-Induced Thrombocytopenia is Linked to Increased Healthcare Utilisation in Glioblastoma and Disproportionally Impacts Female Patients

Abstract Background Thrombocytopenia is a major temozolomide-induced adverse event during the standard treatment of glioblastoma. Consequently, platelet transfusions and treatment modifications may impact quality of life and long-term treatment outcomes. Understanding the impact of thrombocytopenia on healthcare utilisation is crucial to mitigate the need for healthcare resources in glioblastoma patients. Here, we assess the influence of thrombocytopenia-related healthcare among patients diagnosed with glioblastoma. Methods We retrospectively collected patient information treated at the Brain Tumor Center Amsterdam between 2008 and 2021. The occurrence of thrombocytopenia, patient demographics, treatment details and healthcare utilisation data were gathered from patients who received standard glioblastoma treatment. Associations between temporal severity of thrombocytopenia as categorised by the Common Terminology Criteria for Adverse Events, patient characteristics, and healthcare utilisation were analysed using Generalized Linear Mixed Models. Results We included 206 patients with a median age of 58 years, 35.9% were female and we found that thrombocytopenia (any grade) occurred in 61.1% of patients. The occurrence of thrombocytopenia during CRT was associated with increased healthcare utilisation and was largest in females who developed grade 4 thrombocytopenia compared to those who did not develop thrombocytopenia (OR=5.9, p<0.001 in females vs OR=4.4, p<0.001 in males). Grade 4 thrombocytopenia was also associated with heightened healthcare utilisation during the adjuvant phase (OR=7.6, p<0.001), and was comparable between sexes. Conclusions Severe thrombocytopenia during glioblastoma treatment is linked to increased healthcare utilisation, disproportionally impacting females. These data suggest that prevention and early management of thrombocytopenia can reduce healthcare utilisation in patients with glioblastoma.

Nitroaromatic-based triazene prodrugs to target the hypoxic microenvironment in glioblastoma.

Hypoxia is a hallmark of the glioblastoma multiforme microenvironment and represents a promising therapeutic target for cancer treatment. Herein, we report nitroaromatic-based triazene prodrugs designed for selective activation by tumoral endogenous reductases and release of the cytotoxic methyldiazonium ion via a self-immolative mechanism. While compounds bearing a 2-nitrofuran bioreductive group were more efficiently activated by nitroreductases, 4-nitrobenzyl prodrugs 1b, 1d and 1e elicited a more pronounced cytotoxic effect against LN-229 and U-87 MG glioblastoma cell lines under hypoxic conditions when compared to temozolomide (TMZ), the golden standard for glioblastoma treatment. This cytotoxic response aligns with the increased apoptosis levels in LN-229 cells and senescence induction in U-87 MG cells, promoted by prodrugs 1d and 1e, under hypoxic conditions. These results highlight the potential of these hypoxia-activated nitroaromatic-based triazene prodrugs for selective delivery of the cytotoxic methyldiazonium ion and support further optimization to provide a safer alternative for glioblastoma treatment.

Nanoparticle-in-Hydrogel Delivery System for the Sequential Release of Two Drugs.

Glioblastoma is the most common and lethal primary brain tumor. Patients often suffer from tumor- and treatment induced vasogenic edema, with devastating neurological consequences. Intracranial edema is effectively treated with dexamethasone. However, systemic dexamethasone requires large doses to surpass the blood brain barrier in therapeutic quantities, which is associated with significant side effects. The aim of this study was to investigate a biodegradable, dextran-hydroxyethyl methacrylate (dex-HEMA) based hydrogel, containing polymeric micelles loaded with dexamethasone and liposomes encapsulating dexamethasone phosphate for localized and prolonged delivery. Poly(ethylene glycol)-b-poly(N-2-benzoyloxypropyl methacrylamide (mPEG-b-p(HPMA-Bz)) micelles were loaded with dexamethasone and characterized. The dexamethasone micelles, together with dexamethasone phosphate liposomes, were dispersed in an aqueous dex-HEMA solution followed by radical polymerization using a photoinitiator in combination with light. The kinetics and mechanisms of drug release from this hydrogel were determined. The diameter of the nanoparticles was larger than the mesh size of the hydrogel, rendering them immobilized in the polymer network. The micelles immediately released free dexamethasone from the hydrogel for two weeks. The dexamethasone phosphate loaded in the liposomes was not released until the gel degraded and intact liposomes were released, starting after 15 days. The different modes of release result in a biphasic and sequential release profile of dexamethasone followed by dexamethasone phosphate liposomes. The results show that this hydrogel system loaded with both dexamethasone polymeric micelles and dexamethasone phosphate loaded liposomes has potential as a local delivery platform for the sequential release of dexamethasone and dexamethasone phosphate, for the intracranial treatment of glioblastoma associated edema.

Boric acid impedes glioblastoma growth in a rat model: insights from multi-approach analysis

Limited advancements in managing malignant brain tumors have resulted in poor prognoses for glioblastoma (GBM) patients. Standard treatment involves surgery, radiotherapy, and chemotherapy, which lack specificity and damage healthy brain tissue. Boron-containing compounds, such as boric acid (BA), exhibit diverse biological effects, including anticancer properties. This study aimed to examine whether boron supplementation, as BA, can inhibit glioblastoma growth in a xenograft animal model. Using MRI-based tumor size measurement, survival rates, hematological, clinical biochemistry analyses, and genotoxicity parameters, we assessed the impact of BA. Histopathological, immunohistochemical, and immunofluorescence examinations were also conducted. All BA doses (3.25, 6.5, and 13 mg kg−1 b.w.) extended survival compared to GBM controls after 14 days, with a dose-dependent anti-GBM effect observed in MRI analyses. BA treatment improved hematological (WBC and PLT counts) and biochemical parameters (LDL-C, CREA, and ALP). Histopathological examination revealed a significant reduction in tumor diameter with 6.5 and 13 mg kg−1 BA. Immunohistochemical and immunofluorescence staining showed modulation of intracytoplasmic Ki67, cytoplasmic CMPK2, and GFAP expressions in tumor cells post-BA treatment. Additionally, BA did not increase micronuclei formations, indicating its non-genotoxic nature. In conclusion, targeting tumor suppressor networks with boron demonstrates significant therapeutic potential for GBM treatment.

Targeting JAK/STAT3 in glioblastoma cells using an alginate-PNIPAm molecularly imprinted hydrogel for the sustained release of ruxolitinib.

Glioblastoma (GBM) is a notoriously aggressive primary brain tumor characterized by elevated recurrence rates and poor overall survival despite multimodal treatment. Local treatment strategies for GBM are safer and more effective alternatives to systemic chemotherapy, directly tackling residual cancer cells in the resection cavity by circumventing the blood-brain barrier. Molecularly imprinted polymers (MIPs) are promising drug delivery systems due to their high-affinity binding cavities that enable tailored release kinetics. This study reports the development of a semi-synthetic polysaccharide MIP-based hydrogel intended for the post-surgical management of GBM. The biodegradable implant, made of calcium-crosslinked alginate-poly(N-isopropylacrylamide) graft copolymer, was designed for the sustained release of ruxolitinib (RUX) in the resection cavity, targeting the Janus kinase/Signal Transducer and Activator of Transcription-3 signaling pathway. The molecularly imprinted hydrogel demonstrated thermo-thickening and shear-thinning behavior, high entrapment efficiency of RUX (84.59±0.73%), and sustained release over 14days, underscoring the advantages that molecular imprinting of the alginate matrix provides compared to conventional MIPs. The dose-dependent inhibitory effects of the imprinted hydrogel against U251 and A172 GBM cells were demonstrated by increased apoptosis, reduced confluence, colony formation, and delayed wound healing, whereas the non-imprinted hydrogel was biocompatible. The MIP hydrogel could be a safe and effective GBM treatment.

V-ATPase in glioma stem cells: a novel metabolic vulnerability

BackgroundGlioblastoma (GBM) is a lethal brain tumor characterized by the glioma stem cell (GSC) niche. The V-ATPase proton pump has been described as a crucial factor in sustaining GSC viability and tumorigenicity. Here we studied how patients-derived GSCs rely on V-ATPase activity to sustain mitochondrial bioenergetics and cell growth.MethodsV-ATPase activity in GSC cultures was modulated using Bafilomycin A1 (BafA1) and cell viability and metabolic traits were analyzed using live assays. The GBM patients-derived orthotopic xenografts were used as in vivo models of disease. Cell extracts, proximity-ligation assay and advanced microscopy was used to analyze subcellular presence of proteins. A metabolomic screening was performed using Biocrates p180 kit, whereas transcriptomic analysis was performed using Nanostring panels.ResultsPerturbation of V-ATPase activity reduces GSC growth in vitro and in vivo. In GSC there is a pool of V-ATPase that localize in mitochondria. At the functional level, V-ATPase inhibition in GSC induces ROS production, mitochondrial damage, while hindering mitochondrial oxidative phosphorylation and reducing protein synthesis. This metabolic rewiring is accompanied by a higher glycolytic rate and intracellular lactate accumulation, which is not exploited by GSCs for biosynthetic or survival purposes.ConclusionsV-ATPase activity in GSC is critical for mitochondrial metabolism and cell growth. Targeting V-ATPase activity may be a novel potential vulnerability for glioblastoma treatment.

Targeting glycolysis: exploring a new frontier in glioblastoma therapy.

Glioblastoma(GBM) is a highly malignant primary central nervous system tumor that poses a significant threat to patient survival due to its treatment resistance and rapid recurrence.Current treatment options, including maximal safe surgical resection, radiotherapy, and temozolomide (TMZ) chemotherapy, have limited efficacy.In recent years, the role of glycolytic metabolic reprogramming in GBM has garnered increasing attention. This review delves into the pivotal role of glycolytic metabolic reprogramming in GBM, with a particular focus on the multifaceted roles of lactate, a key metabolic product, within the tumor microenvironment (TME). Lactate has been implicated in promoting tumor cell proliferation, invasion, and immune evasion. Additionally, this review systematically analyzes potential therapeutic strategies targeting key molecules within the glycolytic pathway, such as Glucose Transporters (GLUTs), Monocarboxylate Transporters(MCTs), Hexokinase 2 (HK2), 6-Phosphofructo-2-Kinase/Fructose-2,6-Biphosphatase 3 (PFKFB3), Pyruvate Kinase Isozyme Type M2 (PKM2), and the Lactate Dehydrogenase A (LDHA). These studies provide a novel perspective for GBM treatment. Despite progress made in existing research, challenges remain, including drug penetration across the blood-brain barrier, side effects, and resistance. Future research will aim to address these challenges by improving drug delivery, minimizing side effects, and exploring combination therapies with radiotherapy, chemotherapy, and immunotherapy to develop more precise and effective personalized treatment strategies for GBM.

Sodium valproate enhances efficacy of NKG2D CAR-T cells against glioblastoma.

Chimeric antigen receptor T-cell (CAR-T) therapies have shown promise in glioblastoma clinical studies, but responses remain inconsistent due to heterogeneous tumor antigen expression and immune evasion post-treatment. NKG2D CAR-T cells have demonstrated a favorable safety profile in patients with hematologic tumors, and showed robust antitumor efficacy in various xenograft models, including glioblastoma. However, malignant glioma cells evade immunological surveillance by reducing NKG2D ligands expression or cleavage. To enhance the effectiveness of NKG2D CAR-T therapy, we investigated the potential of combining NKG2D CAR-T with approved drugs that cross the blood-brain barrier and augment NKG2D ligands expression in glioma cells. We found that sodium valproate (VPA), an antiepileptic drug, significantly increased surface NKG2D ligands expression on glioblastoma cells at a sublethal concentration. VPA treatment enhanced the susceptibility of glioblastoma cells to NKG2D CAR-T mediated cytotoxicity in both 2D monolayer and 3D tumor spheroid models in vitro. Moreover, VPA-treated glioblastoma cells stimulated CAR-T cells to produce higher levels of inflammatory cytokines (IL-2, IFN-γ, and IL-6). Mechanistically, VPA upregulated NKG2D ligands expression via the PI3K/Akt signaling pathway. Additionally, VPA treatment augmented the antitumor activity of NKG2D CAR-T cells in a glioblastoma xenograft model in vivo. These preclinical results suggest that combining VPA with NKG2D CAR-T therapy represents a promising strategy for improving glioblastoma treatment, warranting further clinical investigation.

Disease stage-specific role of the mitochondrial pyruvate carrier suppresses differentiation in temozolomide and radiation-treated glioblastoma.

The mitochondrial pyruvate carrier (MPC), a central metabolic conduit linking glycolysis and mitochondrial metabolism, is instrumental in energy production. However, the role of the MPC in cancer is controversial. In particular, the importance of the MPC in glioblastoma (GBM) disease progression following standard temozolomide (TMZ) and radiation therapy (RT) remains unexplored. Leveraging in vitro and in vivo patient-derived models of TMZ-RT treatment in GBM, we characterize the temporal dynamics of MPC abundance and downstream metabolic consequences using state-of-the-art molecular, metabolic, and functional assays. Our findings unveil a disease stage-specific role for the MPC, where in post-treatment GBM, but not therapy-naïve tumors, the MPC acts as a central metabolic regulator that suppresses differentiation. Temporal profiling reveals a dynamic metabolic rewiring where a steady increase in MPC abundance favors a shift towards enhanced mitochondrial metabolic activity across patient GBM samples. Intriguingly, while overall mitochondrial metabolism is increased, acetyl-CoA production is reduced in post-treatment GBM cells, hindering histone acetylation and silencing neural differentiation genes in an MPC-dependent manner. Finally, the therapeutic translations of these findings are highlighted by the successful pre-clinical patient-derived orthotopic xenograft (PDOX) trials utilizing a blood-brain-barrier (BBB) permeable MPC inhibitor, MSDC-0160, which augments standard TMZ-RT therapy to mitigate disease relapse and prolong animal survival. Our findings demonstrate the critical role of the MPC in mediating GBM aggressiveness and molecular evolution following standard TMZ-RT treatment, illuminating a therapeutically-relevant metabolic vulnerability to potentially improve survival outcomes for GBM patients.

Laboratory-Engineered Glioblastoma Organoid Culture and Drug Screening.

Glioblastoma (GBM) is described as a group of highly malignant primary brain tumors and stands as one of the most lethal malignancies. The genetic and cellular characteristics of GBM have been a focal point of ongoing research, revealing that it is a group of heterogeneous diseases with variations in RNA expression, DNA methylation, or cellular composition. Despite the wealth of molecular data available, the lack of transferable pre-clinic models has limited the application of this information to disease classification rather than treatment stratification. Transferring the patients' genetic information into clinical benefits and bridging the gap between detailed descriptions of GBM, genotype-phenotype associations, and treatment advancements remain significant challenges. In this context, we present an advanced human GBM organoid model, the Laboratory Engineered Glioblastoma Organoid (LEGO), and illustrate its use in studying the genotype-phenotype dependencies and screening potential drugs for GBM. Utilizing this model, we have identified lipid metabolism dysregulation as a critical milestone in GBM progression and discovered that the microsomal triglyceride transfer protein inhibitor Lomitapide shows promise as a potential treatment for GBM.