Background Chen's U-suture technique was presented with a low incidence of clinically relevant postoperative pancreatic fistula (CR-POPF) in 2014. This study aimed to compare the outcomes of Chen's U-suture technique with those of duct-to-mucosa and traditional invagination pancreaticojejunostomy. Methods The data of patients who underwent pancreaticoduodenectomy across 21 hospitals between 2014 and 2019 were analyzed and categorized into Chen's group, the duct-to-mucosa (DTM) group, and the traditional invagination (TIG) group. Propensity score matching (PSM) analysis was performed to balance the baseline differences among three groups. Subsequently, the surgical outcomes were compared across the groups. Results After PSM, 1060 patients in each group were matched, resulting in balanced baseline characteristics. The CR-POPF rate was 5.19 % in Chen's group, compared to 8.02 % in the TIG group and 7.45 % in the DTM group (P=0.025). A statistically significant difference was identified between Chen's group and the TIG group (P = 0.034), whereas no significant difference was observed when comparing Chen's group to the DTM group (P = 0.060). In the subgroup with a small pancreatic duct (≤3 mm), the CR-POPF rate in Chen's group was significantly lower than that in the DTM group (4.6 % vs 7.4 %, P = 0.019). The incidences of intra-abdominal infection and abscess in Chen's group were 7.45 % and 0.47 % respectively, compared to 10.28 % and 2.26 % in the TIG group, and 10.19 % and 1.51 % in DTM group (all P < 0.05). No significant differences were observed in the rates of severe complications or mortality among the three groups. Conclusions Chen's U-suture technique was a better invagination pancreaticojejunostomy with decreased CR-POPF and POPF-related infection. Furthermore, it was superior to the duct-to-mucosa method for the patients with a small pancreatic duct.

Cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitors are widely used in the treatment of estrogen receptor–positive (ER⁺) breast cancer; however, the metabolic adaptations induced by CDK4/6 inhibition remain incompletely defined. In ER⁺ breast cancer, estrogen signaling plays a central role in coordinating cell cycle progression and metabolic programs that support tumor growth. Glycolytic flux is regulated at the level of phosphofructokinase-1 (PFK1) through the inducible enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3), which is transcriptionally regulated by estrogen receptor signaling and has been shown to promote glycolysis and proliferation in ER⁺ breast cancer cells. Yet, how CDK4/6 inhibition intersects with estrogen-regulated glycolytic control to rewire glucose utilization in ER⁺ breast cancer has not been explored. Glucose metabolism was assessed using extracellular flux analysis, untargeted metabolomics, and stable isotope tracing with uniformly labeled 13C-glucose in ER+ breast cancer cell lines. In vivo metabolic tracing was performed following bolus administration of [U-13C]-glucose. The effects of pharmacologic PFKFB3 inhibition, alone and in combination with CDK4/6 inhibitors, were evaluated in vitro and in patient-derived xenograft (PDX) models. Statistical analyses were performed using appropriate tests with correction for multiple comparisons where applicable. CDK4/6 inhibition increased glycolytic flux, as evidenced by elevated basal and compensatory glycolysis, accumulation of early glycolytic intermediates, and increased 13C labeling of fructose 1,6-bisphosphate. PFKFB3 deficiency significantly attenuated the CDK4/6 inhibitor-induced increase in glycolytic flux. Despite increased glycolysis, stable isotope tracing revealed markedly reduced incorporation of glucose-derived carbon into nucleotide biosynthesis and lipid-associated metabolites, consistent with reduced anabolic demand during G1 cell cycle arrest. In vivo glucose tracing demonstrated a dissociation between increased glycolytic flux and downstream biosynthetic utilization. Pharmacologic inhibition of PFKFB3 imposed additional constraints on glucose utilization and significantly enhanced the antitumor efficacy of CDK4/6 inhibition in PDX models. CDK4/6 inhibition rewires glucose metabolism in ER+ breast cancer by increasing glycolytic flux while limiting downstream glucose utilization, resulting in heightened reliance on regulated glycolytic control to maintain metabolic homeostasis during cell cycle arrest. Disruption of this adaptive metabolic state through PFKFB3 inhibition enhances the antitumor effects of CDK4/6 inhibition and supports the therapeutic potential of targeting glycolytic regulation in combination with CDK4/6 inhibitor-directed therapies.

Lysosomes are critical for maintaining cellular homeostasis and nutrient availability, yet how tumor cells survive under lysosomal inhibition remains unclear. Here, we revealed that inhibiting lysosome function with chloroquine unexpectedly stimulated glucose uptake across various cancer cells. This effect was driven by sterol regulatory element-binding protein 1 (SREBP-1), a key lipogenic transcription factor, which specifically increased the expression of glucose transporters GLUT3 and GLUT6, enhancing glucose uptake and macromolecule synthesis. Elevated glucose, induced by chloroquine, stabilized SREBP cleavage-activating protein (SCAP), the activator of SREBP-1, further amplifying its activity and contributing to tumor resistance to lysosome inhibition. Disrupting this SREBP-1-glucose uptake feedforward loop by combining chloroquine with inhibitors of glucose transporters, SREBP-1, or lipogenic enzymes induced a synergistic antitumor effect in squamous cell and adenocarcinoma lung cancer patient-derived organoids and xenografts. This combination impaired mitochondrial structure and function, inducing apoptotic tumor cell death. Our study uncovers a role for SREBP-1 in regulating glucose metabolism and provides a promising therapeutic strategy that combines lysosome inhibition with glucose transporter or lipogenic enzyme inhibition for effective cancer treatment.

The impact of advanced age on pancreaticoduodenectomy (PD) remains controversial, partly due to inconsistent definitions of “elderly”. To determine the optimal age cut-off for risk stratification and evaluate the safety of different surgical approaches and anastomotic techniques in elderly patients, this study retrospectively analyzed clinical data of 7,028 patients who underwent PD between 2014 and 2019 at 21 centers. Logistic regression analysis demonstrated that advancing age was significantly associated with increased risks of clinically relevant postoperative pancreatic fistula (CR-POPF), bile leakage, pulmonary infection, intra-abdominal infection, and mortality. The optimal age cut-off for predicting CR-POPF was determined as 65 years using receiver operating characteristic curve analysis and the Youden index. Patients were stratified into younger and elderly groups based on this threshold. Both before and after propensity score matching, the elderly group continued to demonstrate significantly higher rates of CR-POPF (before matching: 9.02

Alcohol-associated liver disease (ALD) is associated with disturbances in bile acid (BA) metabolism. Several mouse models have been established to mimic human ALD in the clinical setting for mechanistic investigations, and differences in BA metabolism between these models have not been systematically studied. We quantified BA alterations by liquid chromatography-mass spectrometry (LC-MS) in the livers of two widely used mouse models: the chronic Lieber-DeCarli ethanol diet (CLD) model and the National Institute on Alcohol Abuse and Alcoholism binge-on-chronic alcohol feeding (NIAAA) model, both of which aim to mimic the early stages of human ALD. Statistical analysis showed that total BA levels did not change significantly in either model. However, unconjugated BAs were elevated in both models, and glycol-conjugated BAs were significantly increased only in the NIAAA model. The deconjugation capacity of ursodeoxycholic acid (UDCA) and β-muricholic acid (β-MCA) was increased in the CLD model, whereas that of cholic acid (CA) and lithocholic acid (LCA) was increased in the NIAAA model. NIAAA mice showed increased FXR affinity, implying that the classical biosynthetic pathway of hepatic BAs was inhibited. In conclusion, although total BA levels remained unchanged in the early stages of ALD in both models, the BA composition was more altered in the NIAAA model than in the CLD model, suggesting that different ALD mouse models may exhibit divergent regulatory mechanisms for BA metabolism.

Stable isotopes have frequently been used to study metabolic processes in live cells both in vitro and in vivo. Glutamine, the most abundant amino acid in human blood, plays multiple roles in cellular metabolism by contributing to the production of nucleotides, lipids, glutathione, and other amino acids. It also supports energy production via anaplerosis of tricarboxylic acid cycle intermediates. While 13C-glutamine has been extensively employed to study glutamine metabolism in various cell types, detailed analyses of specific lipids derived from 13C-glutamine via the reductive carboxylation pathway are limited. In this protocol, we present a detailed procedure to investigate glutamine metabolism in human glioblastoma (GBM) cells by conducting 13C-glutamine tracing coupled with untargeted metabolomics analysis using liquid chromatography-mass spectrometry (LC-MS/MS). The method includes step-by-step instructions for the extraction and detection of polar metabolites and long-chain fatty acids (LCFAs) derived from 13C-glutamine in GBM cells. Notably, this approach enables the distinction between isomers of two monounsaturated FAs with identical masses: palmitoleic acid (16:1n-7) (cis-9-hexadecenoic acid) and palmitelaidic acid (16:1n-7) (trans-9-hexadecenoic acid) derived from 13C-glutamine through the reductive carboxylation process. In addition, using this protocol, we also unveil previously unknown metabolic alterations in GBM cells following lysosome inhibition by the antipsychotic drug pimozide. Key features • Methods for analyzing the flux of the stable isotope 13C-glutamine in cancer cells and identifying its derived polar metabolites and long-chain fatty acids (LCFAs). • Distinguishes isomers of long-chain fatty acids, such as palmitoleic acid (16:1n-7) (cis-9-Hexadecenoic acid) and palmitelaidic acid (16:1n-7) (trans-9-Hexadecenoic acid), which share the exact same mass. • The method is utilized to investigate glutamine metabolism reprogramming in cancer cells following lysosome inhibition.

Background/Aims: We explored the possibility of using urine polar metabolites as non-invasive biomarkers of alcohol-associated liver disease (ALD) for early-stage diagnosis and severity assessment, as well as the possible changes in metabolic pathways in ALD patients. Methods: Polar metabolites were extracted with 80% methanol, and parallel 2DLC-MS was used for polar metabolite quantification. Results: Data from untargeted metabolomics showed that 194 metabolites were significantly changed in patients, and three metabolites can differentiate healthy controls (HC), non-severe ALD, and severe alcohol-associated hepatitis (severe AH) with high accuracy (0.92–0.97). Pathway analysis showed that arginine biosynthesis and histidine metabolism pathways were among the pathways containing the metabolites that were most altered in the urine of patients. Metabolites in the urea cycle, histidine catabolism, and histidine dipeptides pathways were notably increased in the urine of ALD patients, but none of the metabolites in these two pathways can simultaneously differentiate patients from healthy volunteers and non-severe ALD from severe AH. As the top differentiated pathways, the alterations of arginine biosynthesis and histidine metabolism indicate their importance in the metabolic dysfunction of ALD. Conclusions: Our results show that the abundance changes of specific metabolites can differentiate the disease severity of ALD, showing the potential of urine polar metabolites as non-invasive biomarkers for early-stage diagnosis and disease severity assessment of ALD.

Elevated hexosamine biosynthesis fuels tumor growth by facilitating protein and lipid glycosylation. But which enzyme in this pathway is better to serve as an antitumor target remains unclear. Here, we revealed that targeting GFAT1, the rate-limiting enzyme in hexosamine synthesis, exhibits limited inhibitory effects on glioblastoma (GBM), the most lethal brain tumor. This outcome is due to the compensation of NAGK-mediated hexosamine salvage pathway. Unexpectedly, inhibiting PGM3, which controls the flux of both de novo hexosamine synthesis and salvage pathways, down-regulates the expression of other enzymes in this pathway and suppresses SREBP-1, a critical lipogenic transcription factor, effectively inhibiting GBM growth. Unexpectedly, SREBP-1 transcriptionally up-regulates the expression of hexosamine synthesis enzymes, while inhibition of these enzymes in turn down-regulates SREBP-1 activation via reducing N-glycosylation of its transporter, SCAP. Our study identified PGM3 as a promising target for treating GBM. Its inhibition disrupts the SREBP-1 activation-hexosamine synthesis positive feedback regulation to effectively eliminate GBM cells.

Background: Wastewater-based epidemiology (WBE) enables the population-level surveillance of molecular and chemical targets. Despite the high prevalence of respiratory diseases, there is a lack of sensitive analytical methods for detecting associated medications in complex wastewater matrices. Methods: We developed and validated a liquid chromatography-mass spectrometry (LC-MS)/MS method using multiple reaction monitoring for 10 common respiratory pharmaceuticals. The workflow integrated freeze-drying for preconcentration, online solid-phase extraction for cleanup, and stable isotope-labeled internal standards (SILs) to compensate for matrix effects. Results: Detection and quantification limits ranged from 0.7 to 19 ng L-1 and 3 to 125 ng L-1, respectively, with recoveries of 82-194% and precision within 0.14-7.2% relative standard deviation. Matrix effects (64-228%) were effectively corrected using SILs. Application to 12 neighborhood-level wastewater samples detected 9 of the 10 target compounds, with 6 (albuterol, amoxicillin, azithromycin, cetirizine, diphenhydramine, and fexofenadine), detected above their quantification limits. Fexofenadine was the most abundant, reaching 3309 ng L-1. Conclusion: This robust, low-volume, high-throughput LC-MS/MS method enables the reliable detection of respiratory pharmaceuticals in wastewater, supporting WBE applications for pharmaceutical use surveillance.

OBJECTIVE:Spatial metabolic differences recently found in glioblastoma (GBM) have been linked to the infiltrating nature of the tumor edge tissue, which is mostly unresectable, and to the tumor core tissue, which resists therapy. The impact of metabolic dysregulation in core and edge GBM tissues on patient survival remains unclear. This study evaluated metabolites obtained from core and edge GBM tissues at the time of resection as biomarkers to risk stratify patients in terms of overall survival (OS). METHODS:Paired core and edge tumor samples from 27 patients with glioma obtained after craniotomy were evaluated postsurgery with high-resolution 2D liquid chromatography-mass spectrometry/mass spectrometry, and metabolomic data for grade IV samples (n = 21) were analyzed by Kaplan-Meier survival analysis and univariable and multivariable Cox proportional hazard regression models. GBM patients were stratified into low- and high-risk groups via a linear equation based on log-transformed signal intensities of key metabolites. Risk scores were generated by summing the product of weights and metabolite signal intensities for each patient's tumor. Weights for significant metabolites were calculated by scaling the univariable Cox proportional hazard ratio for each metabolite by the standard error. For risk score validation, OS events were predicted using an Extreme Gradient Boosting model with Linear Booster (XGBL). RESULTS:Kaplan-Meier survival analysis identified 6 significant metabolites in core tissue and 5 in edge tissue, respectively. Key metabolites in core and edge tissue identified through univariable Cox regression analyses combined with covariates were used to generate multivariable Cox regression models, with edge metabolites remaining significant after correction by patient sex and age at resection. Risk scores based on either 4 core or 11 edge metabolites, or the combination of both, with covariates, generated multivariable Cox regression models significantly associated with OS. Risk score derived from core metabolites remained significant after correction by covariates and was validated with XGBL classification model (area under the receiver operating characteristic curve = 0.876). CONCLUSIONS:OS of patients with GBM can be stratified based on metabolomic differences between core and edge tumor tissues.

Stable isotopes have frequently been used to study metabolic processes in live cells both in vitro and in vivo. Glutamine, the most abundant amino acid in human blood, plays multiple roles in cellular metabolism by contributing to the production of nucleotides, lipids, glutathione, and other amino acids. It also supports energy production via anaplerosis of tricarboxylic acid cycle intermediates. While 13C-glutamine has been extensively employed to study glutamine metabolism in various cell types, detailed analyses of specific lipids derived from 13C-glutamine via the reductive carboxylation pathway are limited. In this protocol, we present a detailed procedure to investigate glutamine metabolism in human glioblastoma (GBM) cells by conducting 13C-glutamine tracing coupled with untargeted metabolomics analysis using liquid chromatography-mass spectrometry (LC-MS/MS). The method includes step-by-step instructions for the extraction and detection of polar metabolites and long-chain fatty acids (LCFAs) derived from 13C-glutamine in GBM cells. Notably, this approach enables the distinction between isomers of two monounsaturated FAs with identical masses: palmitoleic acid (16:1n-7) (cis-9-hexadecenoic acid) and palmitelaidic acid (16:1n-7) (trans-9-hexadecenoic acid) derived from 13C-glutamine through the reductive carboxylation process. In addition, using this protocol, we also unveil previously unknown metabolic alterations in GBM cells following lysosome inhibition by the antipsychotic drug pimozide. Key features • Methods for analyzing the flux of the stable isotope 13C-glutamine in cancer cells and identifying its derived polar metabolites and long-chain fatty acids (LCFAs). • Distinguishes isomers of long-chain fatty acids, such as palmitoleic acid (16:1n-7) (cis-9-Hexadecenoic acid) and palmitelaidic acid (16:1n-7) (trans-9-Hexadecenoic acid), which share the exact same mass. • The method is utilized to investigate glutamine metabolism reprogramming in cancer cells following lysosome inhibition.

PurposeThis study aims to investigate global metabolomic effects of soy protein supplementation.Design/methodology/approachThis study was designed as an exploratory study based on residual specimens from a randomized placebo-controlled trial. The authors investigated changes in serum metabolomic and lipidomic profiles of prostate cancer patients who were assigned to soy protein (19.2 g/day, 41 mg isoflavones, n = 10) or a casein-based placebo (19.8 g/day n = 11) for up to one year following prostatectomy. A total of 63 serum samples collected at baseline, 6 and 12 months were analyzed by untargeted, global metabolomics.FindingsPartial least squares discriminant analysis, based on 493 polar metabolites and 4,706 lipids, showed clear separation of changes in metabolomic profiles between the soy and placebo groups. However, the only metabolite that was significantly different after multiple comparison adjustment was daidzein 4 '-sulfate (p = 0.036). A few other metabolites/lipids were weakly associated with soy supplementation; betaine and C18-ceramide derivative increased and l-alpha-aminoadipic acid and a linolenic acid derivative decreased, compared with the placebo.Originality/valueThe present study demonstrates that physiologically relevant dietary levels of soy protein supplementation close to real-life levels of consumption lead to a shift in global metabolomic profiles.

ABSTRACT Infections with the pathogenic free-living amoebae Naegleria fowleri can lead to life-threatening illnesses including catastrophic primary amebic meningoencephalitis (PAM). Efficacious treatment options for these infections are lacking and the mortality rate remains >95% in the US. Glycolysis is very important for the infectious trophozoite lifecycle stage and inhibitors of glucose metabolism have been found to be toxic to the pathogen. Recently, human enolase 2 (ENO2) phosphonate inhibitors have been developed as lead agents to treat glioblastoma multiforme (GBM). These compounds, which cure GBM in a rodent model, are well-tolerated in mammals because enolase 1 (ENO1) is the predominant isoform used systemically. Here, we describe findings that demonstrate that these agents are potent inhibitors of N. fowleri ENO ( Nf ENO) and are lethal to amoebae. In particular, (1-hydroxy-2-oxopiperidin-3-yl) phosphonic acid (HEX) was a potent enzyme inhibitor (IC 50 value of 0.14 ± 0.04 µM) that was toxic to trophozoites (EC 50 value of 0.21 ± 0.02 µM) while the reported CC 50 was >300 µM. Molecular docking simulation revealed that HEX binds strongly to the active site of Nf ENO with a binding affinity of –8.6 kcal/mol. Metabolomic studies of parasites treated with HEX revealed a 4.5 to 78-fold accumulation of glycolytic intermediates upstream of Nf ENO. Last, nasal instillation of HEX increased longevity of amoebae-infected rodents. Two days after infection, animals were treated for 10 days with 3 mg/kg HEX, followed by one week of observation. At the conclusion of the experiment, eight of 12 HEX-treated animals remained alive (resulting in an indeterminable median survival time) while one of 12 vehicle-treated rodents remained, yielding a median survival time of 10.9 days. Brains of six of the eight survivors were positive for amoebae, suggesting the agent at the tested dose suppressed, but did not eliminate, infection. These findings suggest that HEX is a promising lead for the treatment of PAM.

Antipsychotic drugs have been shown to have antitumor effects but have had limited potency in the clinic. Here, we unveil that pimozide inhibits lysosome hydrolytic function to suppress fatty acid and cholesterol release in glioblastoma (GBM), the most lethal brain tumor. Unexpectedly, GBM develops resistance to pimozide by boosting glutamine consumption and lipogenesis. These elevations are driven by SREBP-1, which we find upregulates the expression of ASCT2, a key glutamine transporter. Glutamine, in turn, intensifies SREBP-1 activation through the release of ammonia, creating a feedforward loop that amplifies both glutamine metabolism and lipid synthesis, leading to drug resistance. Disrupting this loop via pharmacological targeting of ASCT2 or glutaminase, in combination with pimozide, induces remarkable mitochondrial damage and oxidative stress, leading to GBM cell death in vitro and in vivo. Our findings underscore the promising therapeutic potential of effectively targeting GBM by combining glutamine metabolism inhibition with lysosome suppression.

Aim:To explore clinical features and prognosis of hepatocellular carcinoma (HCC) in hepatitis B virus surface antigen (HBsAg)-serocleared patients and identify risk factors associated with postoperative recurrence after curative hepatectomy. Methods:Patients who had undergone initial hepatectomy for HCC from January 2010 through December 2022. Clinicopathological data were compared between HBsAg-seropositive and HBsAg-serocleared patients. Furthermore, risk factors associated with early and late postoperative HCC recurrence (early and late recurrences (ER and LR), respectively) were analyzed for HBsAg-serocleared HCC patients treated by curative hepatectomy. Results:A total of 2184 consecutive patients undergoing initial hepatectomy for HCC were enrolled, including 339 (15.5%) HBsAg-serocleared and 1845 (84.5%) HBsAg-seropositive cases. Tumor characteristics were comparable between the two groups. After curative hepatectomy, the ER rate was lower in the HBsAg-serocleared group than in the HBsAg-seropositive group (16.2% vs 26.3%; p = 0.000). LR rates in the HBsAg-seropositive and HBsAg-serocleared groups were similar (8.3% vs 6.9%, respectively, p = 0.418). Multivariate analysis showed that among HBsAg-serocleared patients, Hong Kong Liver Cancer stage and microvascular invasion were risk factors associated with postoperative ER, while γ-glutamyl transferase level and neutrophil-to-lymphocyte ratio were associated with LR. Conclusion:HBsAg-serocleared and HBsAg-seropositive HCC patients exhibited similar tumor characteristics. Curative hepatectomy-treated HBsAg-serocleared HCC patients experienced a lower ER rate and better short-term (⩽3 years) overall survival (OS) rates than their HBsAg-seropositive counterparts. LR, very late recurrence, and long-term (4-, and 5-year) OS rates were similar between the two groups.

Metabolic dysfunction in the liver represents a predominant feature in the early stages of alcohol-associated liver disease (ALD). However, the mechanisms underlying this are only partially understood. To investigate the metabolic characteristics of the liver in ALD, we did a relative quantification of polar metabolites and lipids in the liver of mice with experimental ALD using untargeted metabolomics and untargeted lipidomics. A total of 99 polar metabolites had significant abundance alterations in the livers of alcohol-fed mice. Pathway analysis revealed that amino acid metabolism was the most affected by alcohol in the mouse liver. Metabolites involved in glycolysis and the TCA cycle were decreased, while glycerol 3-phosphate (G3P) and long-chain fatty acids were increased. Relative quantification of lipids unveiled an upregulation of multiple lipid classes, suggesting that alcohol consumption drives metabolism toward lipid synthesis. Results from enzyme expression and activity detection indicated that the decreased activity of mitochondrial glycerol 3-phosphate dehydrogenase contributed to the disordered metabolism.

Abstract An effective therapy for glioblastoma (GBM), the most lethal brain tumor, is lacking1-11. Several antipsychotic drugs, including pimozide, have recently been shown to have anti-GBM potential 12,13, but the development of tumor resistance diminishes their efficacy. Identifying the mechanisms for such resistance will help repurpose these brain-penetrant drugs for GBM therapy. Here, we found that pimozide inhibits cholesterol and fatty acids (FAs) release via lysosome-mediated lipid droplets (LDs) and lipoprotein hydrolysis, starving GBM cells of these essential building blocks. However, we identified that GBM cells become resistant to pimozide-based therapy due to an elevation of glutamine metabolism. Mechanistically, reductions in cholesterol and the promotion of signaling via sterol regulatory element-binding protein 1 (SREBP-1), a classical lipogenic transcription factor14, stimulate glutamine transporter ASCT2 expression, increasing glutamine uptake and consumption. Enhanced glutamine uptake leads to the release of ammonia that further stimulates SREBP-1 activation, thereby forming a feedforward loop to concurrently increase both glutamine metabolism and lipid synthesis, resulting in drug resistance. Pharmacological targeting of either ASCT2, or glutaminase (GLS) to limit glutaminolysis, abolishes this loop, and in combination with pimozide result in marked killing of GBM cells. Our study demonstrates that combinatorial targeting of glutamine metabolism and lysosomal function is a promising approach to treat GBM, which may be applicable to other aggressive cancers that are also addictive to glutamine and lipids.