NUDT5 promotes the growth,metastasis,and Warburg effect of IDH wild-type glioblastoma multiforme cells by upregulating TRIM47

Objective:To explore the regulatory mechanism of NUDT5 in glioblastoma multiforme(GBM).Methods:GEPIA database was used to predict the expressions of NUDT5 and tripartite motif family proteins 47(TRIM47)in GBM patients.RT-qPCR and Western blot analyses were performed to examine NUDT5 expression in GBM cells.LN-229 cell proliferation,migration as well as invasion were estimated by CCK-8,colony formation,wound healing,and Transwell assays following interference with NUDT5.ECAR assay,L-lactic acid kit,glucose detection kit,and ATP detection kit were applied for the detection of glycolysis-related indexes.Co-immunoprecipitation experiment was carried out to verify the relationship between NUDT5 and TRIM47.Results:GEPIA database showed that NUDT5 expression was significantly increased in GBM patients.Inhibiting the expression of NUDT5 in GBM cells significantly suppressed the viability,proliferation,invasion,migration,and glycolysis of GBM cells.Moreover,TRIM47 was highly expressed in GBM cells and interacted with NUDT5.Overexpression of TRIM47 partially reversed the inhibitory effect of NUDT5 downregulation on the proliferation,metastasis,and glycolysis of GBM cells.Conclusions:NUDT5 promotes the growth,metastasis,and Warburg effect of GBM cells by upregulating TRIM47.Both NUDT5 and TRIM47 can be used as targets for GMB treatment.

Glioblastoma multiforme:Diagnosis, treatment, and invasion

Glioblastoma multiforme(GBM) is an essentially incurable brain tumor, which has been explored for approximately a century. Nowadays, surgical resection, chemotherapy, and radiation therapy are still the standardized therapeutic options. However, due to the intrinsic invasion and metastasis features and the resistance to chemotherapy, the survival rate of glioblastoma patients remains unsatisfactory. To improve the current situation, much more research is needed to provide comprehensive knowledge of GBM. In this review, we summarize the latest updates on GBM treatment and invasion. Firstly, we review the traditional and emerging therapies that have been used for GBM treatment. Given the limited efficiency of these therapies, we further discuss the role of invasion in GBM recurrence and progression, and present current research progress on the mode and mechanisms of GBM invasion.

Nanosensitizers for sonodynamic therapy for glioblastoma multiforme: current progress and future perspectives

Glioblastoma multiforme(GBM) is the most common primary malignant brain tumor, and it is associated with poor prognosis. Its characteristics of being highly invasive and undergoing heterogeneous genetic mutation, as well as the presence of the blood–brain barrier(BBB), have reduced the efficacy of GBM treatment. The emergence of a novel therapeutic method, namely, sonodynamic therapy(SDT), provides a promising strategy for eradicating tumors via activated sonosensitizers coupled with low-intensity ultrasound. SDT can provide tumor killing effects for deep-seated tumors, such as brain tumors. However, conventional sonosensitizers cannot effectively reach the tumor region and kill additional tumor cells, especially brain tumor cells. Efforts should be made to develop a method to help therapeutic agents pass through the BBB and accumulate in brain tumors. With the development of novel multifunctional nanosensitizers and newly emerging combination strategies, the killing ability and selectivity of SDT have greatly improved and are accompanied with fewer side effects. In this review, we systematically summarize the findings of previous studies on SDT for GBM, with a focus on recent developments and promising directions for future research.

Exploring Chemovar-Specific Cannabis Extracts Quantification and Evaluation of Cytotoxic Compounds for Targeting Glioblastoma Multiforme

Glioblastoma Multiforme (GBM) represents one of the most aggressive and metastatic brain tumors, with a dismal success rate of less than three percent after five years, particularly in tumors with active immune checkpoints. This necessitates the development of targeted endogenous agents for precise GBM treatment. Previous experiments utilizing Chemovar Specific Cannabis Extractions (CSCEs), fractionated with polar solvents and quantified using Liquid and Gas Column Chromatography combined with Mass Spectrometry (LC/GCMS), have shown reduced viability and motility in human GBM cell lines. However, the complexity of the botanical substance has hindered the personalization of standard cannabis medicines for GBM due to unknown synergistic effects of multiple compounds. To address this limitation, our study focuses on exposing AM251 cells to chemovar fractions extracted using a non-polar solvent, thereby isolating a broader spectrum of constituents. By employing LC/GCMS in conjunction with Nuclear Magnetic Resonance (NMR), we have identified and quantified nine* compounds present in the non-polar CSCE that exhibit significant efficacy (0.1 μM) in inducing cytotoxicity* in GBM tumor cells. Conversely, the polar fraction in our experiment did not demonstrate efficacy against UM251 cells. The quantification of individual compounds within a cannabis extraction that selectively induces cell death in brain tumors holds promise for guiding future research and facilitating the development of a standardized CSCE for GBM therapy.

Consideration of transmembrane water exchange in pharmacokinetic model significantly improves the accuracy of DCE-MRI in estimating cellular density:A pilot study in glioblastoma multiforme

Transmembrane water exchange(TWE)including transcytolemmal water exchange and transvascular water exchange is involved in many in vivo measurements and makes different contributions to the measuring results.In this study,we focus on the potential influence of TWE on the cell density parameter,intracellular water mole fraction pi,derived by dynamic contrast enhanced-magnetic resonance imaging(DCE-MRI)which has been reported as a technique to characterize perfusion and vascularization of tissues,but its accuracy in measuring cell density(or interstitial space)has been questioned.Sixteen patients with glioblastoma multiforme(GBM)were enrolled since GBM shows strong intratumor heterogeneity in both cell density and TWE.All the subjects were collected with DCE-MRI and apparent diffusion coefficient(ADC)map.The latter was considered as a valid surrogate of cell density.Extended Tofts(eTofts)model considering TWE as infinitely large variables and shutter-speed model(SSM)considering TWE as finite ones were used to fit DCE-MRI data.Monte Carlo(MC)and finite difference(FD)methods were used to simulate the influence of TWE on DCE-MRI-derived pi and ADC,respectively.The eTofts model shows a significant overestimation of pi in comparison with SSM in GBM(P<0.001),which is in accordance with MC simulations,and this overestimation shows dependence on the intra-to-extracellular water exchange rate constant(kio).Significant negative correlations between ADC and SSM-derived pi were found in both voxel-wise analyses(t-test P<0.001,average r=-0.74)and inter-subject comparisons(r=-0.63,P=0.009).But no consistent voxel-wise correlations(P>0.05)and a weaker inter-subject negative correlation(r=-0.56,P=0.02)were found between ADC and eTofts-derived pi.Further experimental and FD results revealed that kio made a limited contribution to ADC values in the physiological kio range in GBM,supporting ADC as a valid biomarker of cell density.These results suggest that the DCE-MRI pharmacokinetic shutter-speed model could significantly improve its accuracy in cell density estimation because of the considering transmembrane water exchange.

The cuproptosis-related gene signature predicts clinical diagnosis,prognosis,and immune microenvironment for glioblastoma multiforme

Background:Glioblastoma multiforme(GBM)is a kind of extremely malignant brain tumor with a poor prognosis.The problem of high recurrence rates and drug resistance has been difficult to solve.Cuproptosis is a newly discovered mode of cell death,which is related to the occurrence and progression of GBM.Methods:We screened TCGA and GEO databases to comprehensively analyze the regulation patterns of cuproptosis in GBM based on cuproptosis-related genes(CRGs).The expression levels,copy number variants(CNV),and mutation of CRGs in GBM were probed.Different regulation patterns of cuproptosis were identified and performed GSVA analysis.Then we constructed a prognostic model based on genes that were differentially expressed in all regulation patterns.The immune cells and tumor microenvironment in different risk groups were compared.Finally,the potential therapeutic drugs were predicted.Results:We identified two key differentially expressed genes under different regulation patterns of cuproptosis and constructed a risk prognostic model.There were significant differences in immune infiltration and drug sensitivity between high and low risk groups.Conclusions:We established a risk prognostic model based on two genes and explored its relationship with immune characteristics,which might help to assess the prognosis and treatment sensitivity to immune and targeted therapy of GBM patients.

Radiation-Induced Spinal Glioblastoma Multiforme: A Rare Complication in the Management of Head and Neck Cancer

Background: Radiation-induced gliomas of the spinal cord are rare late complications of spinal cord irradiation that typically occur in patients treated at younger ages. Aim: Raise awareness of radiation induced high grade gliomas with a case presentation and a review of the literature. Case Presentation: A 50-year-old male with Stage IVA squamous cell carcinoma of the oropharynx was treated with external beam radiotherapy with a complete response. Seven years later, he presented with a cervical spinal cord mass on MRI. An open biopsy was performed. Pathology revealed an intramedullary WHO grade IV astrocytoma, (i.e., glioblastoma multiforme) of the cervical spine that fulfilled the criteria for a radiation-induced malignancy. Conclusions : Review of the literature suggests that radiation-induced gliomas tend to be high grade and may occur at the periphery of an irradiated field. Radiation-induced gliomas of the spinal cord are a serious complication of radiotherapy that may occur in older patients with head and neck cancers, but are so rare that it should not affect treatment decisions.

Biomaterial-based in vitro 3D modeling of glioblastoma multiforme

Adult-onset brain cancers,such as glioblastomas,are particularly lethal.People with glioblastoma multiforme(GBM)do not anticipate living for more than 15 months if there is no cure.The results of conventional treatments over the past 20 years have been underwhelming.Tumor aggressiveness,location,and lack of systemic therapies that can penetrate the blood–brain barrier are all contributing factors.For GBM treatments that appear promising in preclinical studies,there is a considerable rate of failure in phaseⅠandⅡclinical trials.Unfortunately,access becomes impossible due to the intricate architecture of tumors.In vitro,bioengineered cancer models are currently being used by researchers to study disease development,test novel therapies,and advance specialized medications.Many different techniques for creating in vitro systems have arisen over the past few decades due to developments in cellular and tissue engineering.Later-stage research may yield better results if in vitro models that resemble brain tissue and the blood–brain barrier are used.With the use of 3D preclinical models made available by biomaterials,researchers have discovered that it is possible to overcome these limitations.Innovative in vitro models for the treatment of GBM are possible using biomaterials and novel drug carriers.This review discusses the benefits and drawbacks of 3D in vitro glioblastoma modeling systems.

Combination immunotherapy of glioblastoma with dendritic cell cancer vaccines,anti-PD-1 and poly I:C

Glioblastoma(GBM)is a lethal cancer with limited therapeutic options.Dendritic cell(DC)-based cancer vaccines provide a promising approach for GBM treatment.Clinical studies suggest that other immunotherapeutic agents may be combined with DC vaccines to further enhance antitumor activity.Here,we report a GBM case with combination immunotherapy consisting of DC vaccines,anti-programmed death-1(anti-PD-1)and poly I:C as well as the chemotherapeutic agent cyclophosphamide that was integrated with standard chemoradiation therapy,and the patient remained disease-free for 69 months.The patient received DC vaccines loaded with multiple forms of tumor antigens,including mRNA-tumor associated antigens(TAA),mRNA-neoantigens,and hypochlorous acid(HOCl)-oxidized tumor lysates.Furthermore,mRNA-TAAs were modified with a novel TriVac technology that fuses TAAs with a destabilization domain and inserts TAAs into full-length lysosomal associated membrane protein-1 to enhance major histocompatibility complex(MHC)class I and II antigen presentation.The treatment consisted of 42 DC cancer vaccine infusions,26 anti-PD-1 antibody nivolumab administrations and 126 poly I:C injections for DC infusions.The patient also received 28 doses of cyclophosphamide for depletion of regulatory T cells.No immunotherapy-related adverse events were observed during the treatment.Robust antitumor CD4t and CD8t T-cell responses were detected.The patient remains free of disease progression.This is the first case report on the combination of the above three agents to treat glioblastoma patients.Our results suggest that integrated combination immunotherapy is safe and feasible for long-term treatment in this patient.A large-scale trial to validate these findings is warranted.

Prognostic significance of annexin VII expression in glioblastomas multiforme in humans

OBJECT:Glioblastoma multiforme(GBM)is the most common and lethal primary brain tumor in adults.It isnearly uniformly fatal,with a median survival time of approximately l year,despite modem treatment modalities.Nevertheless,a range of survival times exists around this median.Efforts to understand why some patients livelonger or shorter than the average may provide insight into the biology of these neoplasms.The annexin VII(ANX7)gene is located on the human chromosome 10q21,a site long hypothesized to harbor tumor

Role of MRI in Differentiation between Postoperative Tumoral Recurrence and Radiation-Induced Brain Necrosis in Patients of Glioblastoma Multiform

The distinction of radiation-induced brain necrosis (RBN) and recurrent glioblastoma multiform (rGBM) remains a diagnostic challenge due to their similarly on routine follow-up imaging studies and also their clinical manifestations. Our purpose of this review article is to evaluate the role of advanced MR imaging techniques such as Perfusion-weighted imaging (PWI), Diffusion-weighted imaging (DWI) and Magnetic resonance spectroscopy (MRS) in the differentiation of RBN and rGBM and their complications together with our experience and knowledge gained during our neuroimaging practice.

Multimodal Imaging to Delineate Tumor Heterogeneity in Cerebral Gliomas

Purpose: Magnetic resonance imaging (MRI) is the gold standard in visualizing brain tumors and their effects on adjacent structures. However, no reliable information concerning different tumor components and borders between perifocal edema and infiltration areas can be received. The aim of the study was to establish and evaluate a multimodal imaging concept, in order to differentiate different biological tumor components and to determine tumor borders. Materials and Methods: 12 patients with cerebral gliomas (four low and eight high grade) received a “morphological” MRI, a 3D MR spectroscopy and a T2* MR perfusion examination prior to surgery. Data was evaluated by defining different tumor components, which were entitled based upon their multimodal characteristics and histological data. Results: In high grade gliomas different components can be differentiated, which were described as: “true edema”, “cellular proliferation”, “vascular proliferation”, “cellular infiltration”, “tumor” and “necrosis”. In low grade gliomas, four different tumor components were found: “true edema”, “cellular infiltration”, “cellular proliferation” and “tumor”. Conclusion: With the applied multimodal imaging and a novel evaluation concept, it was possible to detect different tumor components, which could be helpful in detecting the optimal sites for tumor biopsy. Especially in morphological “edema appearing” sites, this knowledge could be important for the adaption of tumor resection borders and the planning of radiation therapy. Further studies with more patients and histological correlation are needed.

Tumor antigen and MHC expression in glioma cells for immunotherapeutic interventions

AIM: To investigate the expression of tumor-antigens and major histocompatibility complex(MHC)-machinery components in glioblastoma multiforme cell lines flow cytometry staining methods were applied.METHODS: Ten GBM cell lines(three commercially available: U-87 MG, U-138-MG and GMS-10 as well as seven newly established cell lines from individual patients in low-passages: HROG02, HROG04, HROG05, HROG06, HROG10, HROG13 and HROG17) were analyzed for expression of(Ⅰ) general and(Ⅱ) GBMrelated tumor antigens as well as of(Ⅲ) components of the MHC machinery by flow cytometry.RESULTS: All cell lines expressed MHC class?Ⅰ?with seven out of the ten being HLA-A02 positive. Four of the seven primary cell lines additionally expressedMHC class Ⅱ in a constitutive manner. Of note, after interferon gamma(IFN-γ) treatment, all seven cell lines expressed MHC class Ⅱ. The tumor associated antigens(TAA) EGFR and survivin were expressed at high levels in all cell lines; whereas MART-1, RHAMM, WT-1 and IL-13Rα were expressed by at least half of the cell lines and HER2/neu, MAGE-1 and tyrosinase were expressed only by few cell lines. However, all cell lines expressed at least two of the candidate antigens included into this analysis.CONCLUSION: No obvious differences between commercially available and newly-established cell lines were observed. Thus, the latter in low-passages are interesting for(therapy-) screening and immunotherapeutic strategies.

Parametrization of Survival Measures (Part III) Clinical Evidences in Single Arm Studies with Endpoint of Overall Survival

Many clinical trials have prospective or retrospective data-sets without comparison to the control-group formed by the same cohort as the active one. The measured single arm naturally contains the relevant information, however, in most of the cases, it is impossible to obtain it from the complex survival curve without a reference. In our previous articles [1] [2], we had shown that the self-similar Weibull distribution fits the self-organized biological mechanisms well, and so it is the best option to study the single-arm survival curves, where self-organizing process is actively present. With the Weibull decomposition of the survival curve, we can fit at least two subgroups of patients. The weighted sum of the decomposed fractions could be optimized analytically and determining the best parameters of the components and the best composition ratio of the weighted sum is also possible. In this part of our series of articles, we will show how the method works in a real clinical environment through modulated electro-hyperthermia (mEHT) as a complementary method, applied curatively when no other conventional curative therapies are available. The decomposed function of the non-responding group provides an excellent agreement with the historical controls in pancreatic cancer and non-small-cell-lung-cancer studies. In the case of glioblastoma multiform, the historical missing control from the institute where the treatment was made does not allow a comparison. We used a modified Hardin-Jones-Pauling statistical estimation and had shown in single arm clinical trials for advanced pancreas, non-small cell lung cancer and glioblastoma multiforme, that this estimation is applicable, and it is corresponding with the historical arm and with the non-responding group where this comparison was available.

An update on the molecular biology of glioblastoma,with clinical implications and progress in its treatment

Glioblastoma multiforme(GBM)is the most aggressive and common malig-nant primary brain tumor.Patients with GBM often have poor prognoses,with a median survival of∼15 months.Enhanced understanding of the molecular biology of central nervous system tumors has led to modifications in their classifications,the most recent of which classified these tumors into new categories and made some changes in their nomenclature and grading system.This review aims to give a panoramic view of the last 3 years’findings in glioblastoma characterization,its heterogeneity,and current advances in its treatment.Several molecular parameters have been used to achieve an accurate and personalized characterization of glioblastoma in patients,including epigenetic,genetic,transcriptomic and metabolic features,as well as age-and sex-related patterns and the involvement of several noncoding RNAs in glioblastoma progression.Astrocyte-like neural stem cells and outer radial glial-like cells from the subven-tricular zone have been proposed as agents involved in GBM of IDH-wildtype origin,but this remains controversial.Glioblastoma metabolism is characterized by upregulation of the PI3K/Akt/mTOR signaling pathway,promotion of the gly-colytic flux,maintenance of lipid storage,and other features.This metabolism also contributes to glioblastoma’s resistance to conventional therapies.Tumor heterogeneity,a hallmark of GBM,has been shown to affect the genetic expresion,modulation of metabolic pathways,and immune system evasion.GBM’s aggressive invasion potential is modulated by cell-to-cell crosstalk within the tumor microenvironment and altered expressions of specific genes,such as ANXA2,GBP2,FN1,PHIP,and GLUT3.Nevertheless,the rising number of active clinical trials illustrates the efforts to identify new targets and drugs to treat this malignancy.Immunotherapy is still relevant for research purposes,given the amount of ongoing clinical trials based on this strategy to treat GBM,and neoantigen and nucleic acid-based vaccines are gaining importance due to their antitumoral activity by inducing the immune response.Furthermore,there are clinical trials focused on the PI3K/Akt/mTOR axis,angiogenesis,and tumor heterogeneity for developing molecular-targeted therapies against GBM.Other strategies,such as nanodelivery and computational models,may improve the drug pharmacokinetics and the prognosis of patients with GBM.

Differentiation of radiation necrosis from glioblastoma recurrence after radiotherapy

The standard treatment of glioblastoma,the most common type of primary-brain-tumor,involves radiotherapy with concomitant temozolomide chemotherapy.A patient with glioblastoma,post radiotherapy developed magnatic resonance imaging(MRI)changes consistent with either radiation-induced tumor necrosis or tumor recurrence.Perfusion MRI was suggestive of radiation necrosis,but magnetic resonance spectroscopy and 99mTc-Tetrofosmin single photon emission computed tomography was indicative of tumor recurrence.Positron emission tomography scan was not available.Tumor recurrence was documented by biopsy.Several advanced imaging methods are available to differentiate tumor recurrence from radiation necrosis in glioblastoma patients.However,in inconclusive cases,brain biopsy should be performed for definite diagnosis.

Gene silencing of HIF-2α disrupts glioblastoma stem cell phenotype

malignancy and recurrence in glioblastoma multiforme.Hypoxic regions within the tumor microenvironment help maintain and promote the proliferation of CSCs.Here,we explored the effects of silencing hypoxia inducible factor-2α(HIF-2α)because of its specificity for CSCs within the hypoxic environment.Methods:Cancer stem cell neurospheres were formed by enriching from both the glioblastoma cell line U87 and from brain tumor stem cells isolated directly from human brain tumors.Silencing of human HIF-2αwas performed using both commercial and in-house transfection of a validated short interfering RNA,with all results compared to an established non-silencing control short interfering RNA.Silencing of HIF-2αwas established by Western blotting,and phenotypic effects were assayed by cell migration assays,cell viability measurements,and immunofluorescence staining of differentiation markers.Results:Transfection with either our previously reported pH-sensitive,cationic amphiphilic macromolecule-based delivery system or Lipofectamine was similarly effective in silencing HIF-2α.The chemotherapeutic resistance and neurosphere formation were reduced when HIF-2αwas silenced.Migratory capacities in the presence of macrophage conditioned media were modulated.HIF-2αsilencing was complementary to temozolomide treatment in producing phenotypic rather than cytotoxic effects.Conclusion:HIF-2αsilencing under hypoxia inhibited CSC phenotypes while promoting differentiated cell phenotypes and is complementary to existing DNA alkylating treatments in inhibiting glioma CSC activity.

Integrins and focal adhesion kinase in the malignant behavior of gliomas

Glioblastoma multiforme(GBM)is the most common type of glioma and is associated with a very poor prognosis.The standard treatment includes radiotherapy concurrent with temozolomide,however recently the Food and Drug Administration approved bevacizumab for use in patients with progressive glioblastoma following prior therapy.The limited number of treatment options points to the need for novel effective therapeutic approaches.A promising approach is the use of tyrosine kinase inhibitors(TKIs)in GBM treatment.However,the results from the majority of clinical trials using TKIs are not very encouraging.One growing area is the development of tumor-homing peptides that resemble the integrin recognition sequence RGD.In this article,the role of integrins and focal adhesion kinase in malignant glioma is reviewed,and an experimental study is proposed that will apply a strategy for peptide-mediated delivery of compounds deep into tumor parenchyma using tumor-homing peptides.

A distinction of gliomas at cellular and tissue level by surface-enhanced Raman scattering spectroscopy

Glioblastoma multiforme(GBM)is the most common malignant primary brain tumor in adults.The precise identification and distinction of GBM heterogeneity from surrounding brain parenchyma at the cellular level and even at the tissue level are important for GBM therapy.In this study,GBM cells are distinguished from normal astrocytes and non-central nervous system(CNS)tumor cells by surface-enhanced Raman scattering(SERS)based on gold nanoshell(SiO_(2)@Au)particles and support vector machine(SVM)algorithm.In addition,the gold nanoisland(AuNI)SERS substrates are further developed and explored for accurate detection of GBM at the tissue level.The distinction between glioma and trauma tissues,identification of different tumor grades,and IDH mutation are realized with the assistance of orthogonal partial least squares discriminant analysis(OPLS-DA)in a rapid,non-invasive,and convenient method.The results show that the developed SERS-based analytical method has the potential for practical application for the detection of GBM at the single-cell and tissue levels and even for real-time intraoperative diagnosis.