Melatonin promotes cell cycle progression of neural stem cells subjected to manganese via Nurr1.

Excessive exposure to manganese (Mn) through drinking water and food during pregnancy significantly heightens the likelihood of neurodevelopmental damage in offspring. Multiple studies have indicated that melatonin (Mel) may help to relieve neurodevelopmental disorders caused by Mn, but potential mechanisms underlying this effect require further exploration. Here, we utilized primary neural stem cells (NSCs) as a model to elucidate the molecular mechanism underlying the protective function of Mel on Mn-induced cell proliferation dysfunction and cycle arrest. Our results showed that Mn disrupted the cell cycle in NSCs by suppressing positive regulatory proteins (CDK2, Cyclin A, Cyclin D1, and E2F1) and enhancing negative ones (p27KIP1 and p57KIP2), leading to cell proliferation dysfunction. Mel inhibited the Mn-dependent changes to these proteins and the cell cycle through nuclear receptor-related protein 1 (Nurr1), thus alleviating the proliferation dysfunction. Knockdown of Nurr1 using lentivirus-expressed shRNA in NSCs resulted in a diminished protective effect of Mel. We concluded that Mel mitigated Mn-induced proliferation dysfunction and cycle arrest in NSCs through Nurr1.

SLFN11 promotes clear cell renal cell carcinoma progression via the PI3K/AKT signaling pathway.

SLFN11 is abnormally expressed and associated with survival outcomes in various human cancers. However, the role of SLFN11 in clear cell renal cell carcinoma (ccRCC) remains unclear. This study aimed to investigate the clinical value and potential functions of SLFN11 in ccRCC. Comprehensive bioinformatics analyses were performed using online databases. Quantitative real-time PCR (qPCR) and western blotting were used to validate the expression data. CCK8, flow cytometry analysis, and EdU staining were performed to determine the level of cell proliferation. Flow cytometry analysis was also used to detect cell apoptosis. Wound-healing assay and Transwell assays were performed to assess cell migration and invasion capability, respectively. SLFN11 was overexpressed and was an independent prognostic factor in ccRCC. SLFN11 knockdown inhibited cell proliferation, migration, and invasion and promoted apoptosis. Functional and pathway enrichment analyses suggested that SLFN11 may have an impact on tumorigenesis in ccRCC through regulation of the inflammatory response, the PI3K/AKT signaling pathway and other effectors. Furthermore, SLFN11 knockdown inhibited the phosphorylation of the PI3K/AKT signaling pathway and could be activated by 740 Y-P. Finally, we demonstrated that miR-183 may specifically target SLFN11, and miR-183 expression was correlated with predicted survival. SLFN11 may play a critical role in ccRCC progression and may serve as a novel prognostic biomarker in ccRCC.

Cr(VI) reduction by Agrobacterium sp. Cr-1 and Lysinibacillus sp. Cr-2, novel Cr(VI)-reducing strains isolated from chromium plant soil.

The bioremediation of Cr(VI)-contaminated soil is a promising strategy; however, the performance of Cr(VI)-reducing bacteria is limited by the toxicity of Cr(VI). In this study, two novel Cr(VI)-reducing bacteria were isolated from a Cr salt plant and identified as Agrobacterium sp. and Lysinibacillus sp. The Cr(VI) reduction conditions of the two strains were optimized. At a Cr(VI) concentration of 500 mg/L, Agrobacterium sp. Cr-1 reduced Cr(VI) with a removal rate of 96.91%, while that for Lysinibacillus sp. Cr-2 was 92.82%. First-order reaction kinetic equations simulated the positive relationship between time and Cr(VI) concentration during Cr(VI) reduction in these two strains. Agrobacterium sp. Cr-1 was further studied, and the effects of different cell components on Cr(VI) reduction were detected. The extracellular extracts of Agrobacterium sp. Cr-1 played a major role in Cr(VI) reduction, followed by intracellular extracts and cell membranes. The scanning electron microscope-energy dispersive spectrometer (SEM-EDS) images show that the precipitation was Cr. The high Cr(VI) reducing ability of Agrobacterium sp. Cr-1 suggests that this strain is promising for the remediation of Cr(VI)-contaminated sites.

A liposomal carbohydrate vaccine, adjuvanted with an NKT cell agonist, induces rapid and enhanced immune responses and antibody class switching.

BACKGROUND: Congenital disorders of glycosylation (CDGs) are genetic diseases caused by gene defects in glycan biosynthesis pathways, and there is an increasing number of patients diagnosed with CDGs. Because CDGs show many different clinical symptoms, their accurate clinical diagnosis is challenging. Recently, we have shown that liposome nanoparticles bearing the ALG1-CDG and PMM2-CDG biomarkers (a tetrasaccharide: Neu5Ac-α2,6-Gal-ß1,4-GlcNAc-ß1,4-GlcNAc) stimulate a moderate immune response, while the generated antibodies show relatively weak affinity maturation. Thus, mature antibodies with class switching to IgG are desired to develop high-affinity antibodies that may be applied in medical applications. RESULTS: In the present study, a liposome-based vaccine platform carrying a chemoenzymatic synthesized phytanyl-linked tetrasaccharide biomarker was optimized. The liposome nanoparticles were constructed by dioleoylphosphatidylcholine (DOPC) to improve the stability and immunogenicity of the vaccine, and adjuvanted with the NKT cell agonist PBS57 to generate high level of IgG antibodies. The results indicated that the reformulated liposomal vaccine stimulated a stronger immune response, and PBS57 successfully induce an antibody class switch to IgG. Further analyses of IgG antibodies elicited by liposome vaccines suggested their specific binding to tetrasaccharide biomarkers, which were mainly IgG2b isotypes. CONCLUSIONS: Immunization with a liposome vaccine carrying a carbohydrate antigen and PBS57 stimulates high titers of CDG biomarker-specific IgG antibodies, thereby showing great potential as a platform to develop rapid diagnostic methods for ALG1-CDG and PMM2-CDG.

Evaluation of Xpert MTB/RIF assay for the diagnosis of extrapulmonary tuberculosis in Southwest China.

The purpose of this study was to determine the diagnostic efficacy of Xpert MTB/RIF assay for rapid diagnosis of Tuberculosis (TB) and detection of rifampicin (RIF) resistance in patients suspected of having EPTB, assessing it against traditional culture and drug susceptibility test (DST) by proportional method, and the ability to predict multidrug resistance TB by Xpert MTB/RIF assay. In this study, the Xpert MTB/RIF assay was applied to 1,614 extrapulmonary specimens. Compared with TB culture and Composite Reference Standard (CRS), the Xpert MTB/RIF assay had a high sensitivity and specificity for detection of EPTB. Depending on the culture method or CRS as the standard, sensitivity of the Xpert MTB/RIF assay for detection of MTB in pleural effusion, cerebrospinal fluid, thoracic drainage fluid and throat swabs specimens were lower than that of other specimens. According to the experimental results, we have reason to believe that Xpert MTB/RIF assay is a rapid and simple technique with high sensitivity and specificity for diagnosing EPTB and detecting drug resistance in variety of specimens. Xpert MTB/RIF assay combined with DST maybe identify more cases of multi-drug resistant tuberculosis (MDR-TB).

Crystal Structure, Chemical Bond, and Microwave Dielectric Properties of Ba1-xSrx(Zn1/3Nb2/3)O3 Solid Solution Ceramics.

Ba1-xSrx(Zn1/3Nb2/3)O3 (BSZN) perovskite ceramics are prepared using the traditional solid-state reaction method. X-ray diffraction (XRD), Scanning electron microscopy (SEM), Raman spectroscopy, and X-ray photoelectron spectroscopy (XPS) were used to analyze the phase composition, crystal structure, and chemical states of BSZN ceramics, respectively. In addition, the dielectric polarizability, octahedral distortion, complex chemical bond theory, and PVL theory were investigated in detail. Systematic research showed that Sr2+ addition could considerably optimize the microwave dielectric properties of BSZN ceramics. The change in τf value in the negative direction was attributed to oxygen octahedral distortion and bond energy (Eb), and the optimal value of 1.26 ppm/°C was obtained at x = 0.2. The ionic polarizability and density played a decisive role in the dielectric constant, achieving a maximum of 45.25 for the sample with x = 0.2. The full width at half-maximum (FWHM) and lattice energy (Ub) jointly contributed to improving the Q × f value, and a higher Q × f value corresponded to a smaller FWHM value and a larger Ub value. Finally, excellent microwave dielectric properties (εr = 45.25, Q × f = 72,704 GHz, and τf = 1.26 ppm/°C) were obtained for Ba0.8Sr0.2(Zn1/3Nb2/3)O3 ceramics sintered at 1500 °C for 4 h.

Research Progress of Ba(Zn1/3Nb2/3)O3 Microwave Dielectric Ceramics: A Review.

Ba(Zn1/3Nb2/3)O3 (BZN) microwave dielectric ceramics have attracted great attention due to their high-quality factor (Q), near-zero temperature coefficient of resonant frequency (τf), and suitable dielectric constant (εr), making them promising materials for application in microwave devices. Due to their superior dielectric properties, composite perovskite ceramics are widely used in the field of microwave communication, base stations, navigation, radar, etc. This article summarized the latest research progress of BZN ceramics and discusses the main preparation methods and performance modifications. Furthermore, the problems faced by BZN ceramics and solutions to improve their performance, as well as their potential applications, are analyzed. This article provides a reference for the design and preparation of BZN ceramics.

Simultaneous treatment of chromium-containing wastewater and electricity generation using a plant cathode-sediment microbial fuel cell: investigation of associated mechanism and influencing factors.

A novel plant cathode-sediment microbial fuel cell (P-SMFC) was constructed to treat Cr-containing wastewater, and the effects of the plants used, initial concentrations of Cr(VI) employed, and the external resistance on the treatment of wastewater and generation of electricity were investigated. The results showed that the system achieved the best performance when Acorus calamus was the cathode plant, the external resistance was 2000 Ω, and the initial Cr (VI) concentration of the overlying water of is 230 mg/L. A maximum power density of 40.16 mW/m2 was reached, and Cr (VI) and COD removal efficiencies in the overlying water were 99.94% and 98.21%, respectively. The closed-circuit installation promoted the attachment of many microorganisms to the cathode, anode and sediment, increased species abundance, and reduced species diversity. The P-SMFC is inexpensive to construct, it consumes no energy, and it can generate bioelectricity; it thus has great application development value as a chromium-containing wastewater treatment method.

Crystal Structures and Microwave Dielectric Properties of Novel MgCu2Nb2O8 Ceramics Prepared by Two-Step Sintering Technique.

In this work, novel MgCu2Nb2O8 (MCN) ceramics were synthesized by the two-step sintering (TSS) technique, and the phase composition, crystal structures, and microwave dielectric properties were comprehensively studied. X-ray diffraction (XRD) and Raman analysis demonstrated that MCN ceramics are multi-phase ceramics consisting of MgNb2O6 and CuO phases. X-ray photoelectron spectroscopy (XPS) was utilized to investigate the chemical composition and element valence of MgCu2Nb2O8 ceramics. Scanning electron microscopy (SEM) analysis demonstrated dense microstructures in the MCN ceramics prepared at a sintering temperature of 925 °C. The microwave dielectric properties were largely affected by the lattice vibrational modes and densification level of the ceramics. The outstanding microwave dielectric properties of εr = 17.15, Q × f = 34.355 GHz, and τf = -22.5 ppm/°C were obtained for the MCN ceramics sintered at 925 °C, which are results that hold promise for low temperature co-fired ceramic (LTCC) applications.

Effect of Electrode Distances on Remediation of Eutrophic Water and Sediment by Sediment Microbial Fuel Cell Coupled Floating Beds.

Efficient and sustainable technologies for cleaning of contaminated water and sediments are in urgent demand. In this study, a new type of sediment microbial fuel cell coupled floating bed (FB-SMFC) was developed to repair eutrophic water and sediment in a cleaner way. The effect of electrode spacing on the power generation capacity and the synchronous remediation of pollutants from eutrophic water and sediment were studied. When the electrode distance was 60 cm, the maximum power generation and pollutant removal effects were obtained. At the end of the experiment, the maximum output voltage was 0.4 V, and the chemical oxygen demand (CODCr, potassium dichromate method), total nitrogen (TN), and total phosphorus (TP) contents in the overlying water were 8 mg/L, 0.7 mg/L, and 0.39 mg/L. The corresponding removal rates were 88.2%, 78.8%, and 59.0%, respectively. The removal rates of organic matter and TN in the sediment were 12.8% and 86.4%, respectively, and the fixation rate of TP was 29.2%. Proteobacteria was the dominant phylum of bacteria in the sediment and anode. Many anaerobic bacteria were found in the overlying water, which facilitated denitrification. Overall, the results of this research revealed a highly efficient and reliable strategy for eutrophic water and sediment remediation, aquatic ecosystems restoration, and human health protection.

PTEN loss drives resistance to the neddylation inhibitor MLN4924 in glioblastoma and can be overcome with TOP2A inhibitors.

BACKGROUND: Neddylation inhibition, affecting posttranslational protein function and turnover, is a promising therapeutic approach to cancer. We report vulnerability to MLN4924 or pevonedistat (a neddylation inhibitor) in a subset of glioblastoma (GBM) preclinical models and identify biomarkers, mechanisms, and signatures of differential response. METHODS: GBM sequencing data were queried for genes associated with MLN4924 response status; candidates were validated by molecular techniques. Time-course transcriptomics and proteomics revealed processes implicated in MLN4924 response. RESULTS: Vulnerability to MLN4924 is associated with elevated S-phase populations, re-replication, and DNA damage. Transcriptomics and shotgun proteomics depict PTEN signaling, DNA replication, and chromatin instability pathways as significant differentiators between sensitive and resistant models. Loss of PTEN and its nuclear functions is associated with resistance to MLN4924. Time-course proteomics identified elevated TOP2A in resistant models through treatment. TOP2A inhibitors combined with MLN4924 prove synergistic. CONCLUSIONS: We show that PTEN status serves as both a novel biomarker for MLN4924 response in GBM and reveals a vulnerability to TOP2A inhibitors in combination with MLN4924.

MicroRNA-4521 targets hepatoma up-regulated protein (HURP) to inhibit the malignant progression of breast cancer.

Breast cancer (BC) is a common malignancy among women, and microRNAs (miRNAs) play a role in its progression. Reportedly, microRNA-4521 (miR-4521) participates in regulating the progression of some carcinomas. In the present work, miR-4521 and hepatoma up-regulated protein (HURP) mRNA expressions in BC tissues and cell lines were examined by qRT-PCR. After miR-4521 was overexpressed or knocked down in BC cells, CCK-8 experiment and EdU experiment were employed to detect BC cell growth. Moreover, the Transwell assay was adopted to detect the migration and invasion. Subsequently, flow cytometry was executed to evaluate the changes in cell cycle progression. KEGG analysis was utilized to predict the signaling pathways related with the target genes of miR-524-5p. The binding relationship between miR-4521 and HURP 3'UTR was validated by dual-luciferase reporter gene experiment. HURP and NF-kB p65 protein expression in BC cells was detected by Western blot. It was revealed that, miR-4521 was lowly expressed in BC tissues, and its expression was associated with tumor grade, lymph node metastasis and clinical stage of the patients. Overexpression of miR-4521 suppressed the growth, migration, invasion and cell cycle progression of BC cells and restrained p65 expression; downregulation of miR-4521 in BC cells showed opposite biological effects. Additionally, HURP was a downstream target gene of miR-4521, and overexpression of HURP reversed the above effects of miR-4521 overexpression. In short, miR-4521 can inhibit BC progression by modulating the HURP/NF-κB pathway.

[Research on the influence of mental fatigue on information resources allocation of working memory].

Mental fatigue is the subjective state of people after excessive consumption of information resources. Its impact on cognitive activities is mainly manifested as decreased alertness, poor memory and inattention, which is highly related to the performance after impaired working memory. In this paper, the partial directional coherence method was used to calculate the coherence coefficient of scalp electroencephalogram (EEG) of each electrode. The analysis of brain network and its attribute parameters was used to explore the changes of information resource allocation of working memory under mental fatigue. Mental fatigue was quickly induced by the experimental paradigm of adaptive N-back working memory. Twenty-five healthy college students were randomly recruited as subjects, including 14 males and 11 females, aged from 20 to 27 years old, all right-handed. The behavioral data and resting scalp EEG data were collected simultaneously. The results showed that the main information transmission pathway of the brain changed under mental fatigue, mainly in the frontal lobe and parietal lobe. The significant changes in brain network parameters indicated that the information transmission path of the brain decreased and the efficiency of information transmission decreased significantly. In the causal flow of each electrode and the information flow of each brain region, the inflow of information resources in the frontal lobe decreased under mental fatigue. Although the parietal lobe region and occipital lobe region became the main functional connection areas in the fatigue state, the inflow of information resources in these two regions was still reduced as a whole. These results indicated that mental fatigue affected the information resources allocation of working memory, especially in the frontal and parietal regions which were closely related to working memory.

Elevated fibroblast growth factor-inducible 14 expression transforms proneural-like gliomas into more aggressive and lethal brain cancer.

High-grade gliomas (HGGs) are aggressive, treatment-resistant, and often fatal human brain cancers. The TNF-like weak inducer of apoptosis (TWEAK)/fibroblast growth factor-inducible 14 (Fn14) signaling axis is involved in tissue repair after injury and constitutive signaling has been implicated in the pathogenesis of numerous solid cancers. The Fn14 gene is expressed at low levels in the normal, uninjured brain but is highly expressed in primary isocitrate dehydrogenase wild-type and recurrent HGGs. Fn14 signaling is implicated in numerous aspects of glioma biology including brain invasion and chemotherapy resistance, but whether Fn14 overexpression can directly promote tumor malignancy has not been reported. Here, we used the replication-competent avian sarcoma-leukosis virus/tumor virus A system to examine the impact of Fn14 expression on glioma development and pathobiology. We found that the sole addition of Fn14 to an established oncogenic cocktail previously shown to generate proneural-like gliomas led to the development of highly invasive and lethal brain cancer with striking biological features including extensive pseudopalisading necrosis, constitutive canonical and noncanonical NF-κB pathway signaling, and high plasminogen activator inhibitor-1 (PAI-1) expression. Analyses of HGG patient datasets revealed that high human PAI-1 gene (SERPINE1) expression correlates with shorter patient survival, and that the SERPINE1 and Fn14 (TNFRSF12A) genes are frequently co-expressed in bulk tumor tissues, in tumor subregions, and in malignant cells residing in the tumor microenvironment. These findings provide new insights into the potential importance of Fn14 in human HGG pathobiology and designate both the NF-κB signaling node and PAI-1 as potential targets for therapeutic intervention. MAIN POINTS: This work demonstrates that elevated levels of the TWEAK receptor Fn14 in tumor-initiating, neural progenitor cells leads to the transformation of proneural-like gliomas into more aggressive and lethal tumors that exhibit constitutive NF-κB pathway activation and plasminogen activator inhibitor-1 overexpression.

Uncertainty quantification in the radiogenomics modeling of EGFR amplification in glioblastoma.

Radiogenomics uses machine-learning (ML) to directly connect the morphologic and physiological appearance of tumors on clinical imaging with underlying genomic features. Despite extensive growth in the area of radiogenomics across many cancers, and its potential role in advancing clinical decision making, no published studies have directly addressed uncertainty in these model predictions. We developed a radiogenomics ML model to quantify uncertainty using transductive Gaussian Processes (GP) and a unique dataset of 95 image-localized biopsies with spatially matched MRI from 25 untreated Glioblastoma (GBM) patients. The model generated predictions for regional EGFR amplification status (a common and important target in GBM) to resolve the intratumoral genetic heterogeneity across each individual tumor-a key factor for future personalized therapeutic paradigms. The model used probability distributions for each sample prediction to quantify uncertainty, and used transductive learning to reduce the overall uncertainty. We compared predictive accuracy and uncertainty of the transductive learning GP model against a standard GP model using leave-one-patient-out cross validation. Additionally, we used a separate dataset containing 24 image-localized biopsies from 7 high-grade glioma patients to validate the model. Predictive uncertainty informed the likelihood of achieving an accurate sample prediction. When stratifying predictions based on uncertainty, we observed substantially higher performance in the group cohort (75% accuracy, n = 95) and amongst sample predictions with the lowest uncertainty (83% accuracy, n = 72) compared to predictions with higher uncertainty (48% accuracy, n = 23), due largely to data interpolation (rather than extrapolation). On the separate validation set, our model achieved 78% accuracy amongst the sample predictions with lowest uncertainty. We present a novel approach to quantify radiogenomics uncertainty to enhance model performance and clinical interpretability. This should help integrate more reliable radiogenomics models for improved medical decision-making.

Remediation of soil contaminated with high levels of hexavalent chromium by combined chemical-microbial reduction and stabilization.

In order to solve the problem of re-oxidation after chemical remediation of soil contaminated with high levels of hexavalent chromium (Cr(VI)), we investigated the use of chemical reduction combined with microbial stabilization to remediate soils contaminated with high Cr(VI) concentration. The leaching toxicity and microbial diversity of Cr(VI)-contaminated soil and the leaching toxicity of remediated soil oxidized by potassium permanganate (KMnO4) were measured. The results indicate that the conversion rate of Cr(VI) reached 97 %, and the concentration of Cr(VI) in toxic solutions leaching can be reduced by 95 % after 40 days of microbial stabilization. Sterilization experiments showed that the reduction of Cr(VI) by microorganisms is stable. The results of microbial diversity analysis indicate that bacterial community changed more than fungal community during the reduction process of Cr(VI), and the species abundance and species evenness of bacteria decreased. Bacillus spp. and Halomonas spp. were the dominant species in this study.

Single-Cell RNA Sequencing of a Postmenopausal Normal Breast Tissue Identifies Multiple Cell Types That Contribute to Breast Cancer.

The human breast is composed of diverse cell types. Studies have delineated mammary epithelial cells, but the other cell types in the breast have scarcely been characterized. In order to gain insight into the cellular composition of the tissue, we performed droplet-mediated RNA sequencing of 3193 single cells isolated from a postmenopausal breast tissue without enriching for epithelial cells. Unbiased clustering analysis identified 10 distinct cell clusters, seven of which were nonepithelial devoid of cytokeratin expression. The remaining three cell clusters expressed cytokeratins (CKs), representing breast epithelial cells; Cluster 2 and Cluster 7 cells expressed luminal and basal CKs, respectively, whereas Cluster 9 cells expressed both luminal and basal CKs, as well as other CKs of unknown specificity. To assess which cell type(s) potentially contributes to breast cancer, we used the differential gene expression signature of each cell cluster to derive gene set variation analysis (GSVA) scores and classified breast tumors in The Cancer Gene Atlas (TGGA) dataset (n = 1100) by assigning the highest GSVA scoring cell cluster number for each tumor. The results showed that five clusters (Clusters 2, 3, 7, 8, and 9) could categorize >85% of breast tumors collectively. Notably, Cluster 2 (luminal epithelial) and Cluster 3 (fibroblast) tumors were equally prevalent in the luminal breast cancer subtypes, whereas Cluster 7 (basal epithelial) and Cluster 9 (other epithelial) tumors were present primarily in the triple-negative breast cancer (TNBC) subtype. Cluster 8 (immune) tumors were present in all subtypes, indicating that immune cells may contribute to breast cancer regardless of the subtypes. Cluster 9 tumors were significantly associated with poor patient survival in TNBC, suggesting that this epithelial cell type may give rise to an aggressive TNBC subset.

Temporospatial genomic profiling in glioblastoma identifies commonly altered core pathways underlying tumor progression.

BACKGROUND: Tumor heterogeneity underlies resistance and disease progression in glioblastoma (GBM), and tumors most commonly recur adjacent to the surgical resection margins in contrast non-enhancing (NE) regions. To date, no targeted therapies have meaningfully altered overall patient survival in the up-front setting. The aim of this study was to characterize intratumoral heterogeneity in recurrent GBM using bulk samples from primary resection and recurrent samples taken from contrast-enhancing (EN) and contrast NE regions. METHODS: Whole exome and RNA sequencing were performed on matched bulk primary and multiple recurrent EN and NE tumor samples from 16 GBM patients who received standard of care treatment alone or in combination with investigational clinical trial regimens. RESULTS: Private mutations emerge across multi-region sampling in recurrent tumors. Genomic clonal analysis revealed increased enrichment in gene alterations regulating the G2M checkpoint, Kras signaling, Wnt signaling, and DNA repair in recurrent disease. Subsequent functional studies identified augmented PI3K/AKT transcriptional and protein activity throughout progression, validated by phospho-protein levels. Moreover, a mesenchymal transcriptional signature was observed in recurrent EN regions, which differed from the proneural signature in recurrent NE regions. CONCLUSIONS: Subclonal populations observed within bulk resected primary GBMs transcriptionally evolve across tumor recurrence (EN and NE regions) and exhibit aberrant gene expression of common signaling pathways that persist despite standard or targeted therapy. Our findings provide evidence that there are both adaptive and clonally mediated dependencies of GBM on key pathways, such as the PI3K/AKT axis, for survival across recurrences.