bims-glucam 2026-06-21 papers

bims-glucam

Biomed News

on Glutamine cancer metabolism

Issue of 2026–06–21
fifteen papers selected by
Sreeparna Banerjee, Middle East Technical University

PYCR1 promotes glutamine metabolism and the progression of lung adenocarcinoma by regulating the expression of OPLAH.
Metabolic Reprogramming and Molecular Mechanisms in Renal Cell Carcinoma: Therapeutic Implications and Future Perspectives.
Semaglutide targets muscle mitochondria to regulate glutamine metabolism and treat osteoarthritis.
Posttranscriptional Regulation of Metabolism in Glioblastoma: A Multipathway Review.
Glutamine metabolism shapes the phenotypic adaptation of uterine dendritic cells in response to early allogeneic pregnancy.
The metabolic escape: how tumor metabolic reprogramming drives drug resistance.
Metabolic reprogramming of glutamine is associated with M2 macrophage polarization in allergic rhinitis.
The competition of amino acid among tumors, tumor-associated macrophages and T cells based on metabolic reprogramming.
Ketogenic diet as a metabolic vehicle enhancing the therapeutic efficacy of mebendazole and devimistat in juvenile syngeneic high-grade glioma.
Glutamine Tautomerization Drives RhoGAP-Aided GTP Hydrolysis in Small Rho GTPases.
Protective Effects of Glycyl-Glutamine Dipeptide on In Vitro Brain Ischemia Reperfusion Model and the Role of Sigma-1 Receptors.
Growth-mode-independent glutamine/glutamate/aspartate metabolic shift and urea cycle remodeling driven by FOXA1 in benzo[a]pyrene-transformed human bronchial epithelial cells.
Development of new liquid chromatography-tandem mass spectrometry methods for the quantification of seven metabolites in three related breast cancer cell lines.
Targeted LC-MS/MS profiling of serum amino acids reveals type 2 diabetes-specific alterations in advanced gastric cancer.
Evaluation of FASP for Mass Spectrometry-Based Untargeted Metabolomics Analysis of Urine Samples.

Clin Exp Med. 2026 Jun 15.

PYCR1 promotes glutamine metabolism and the progression of lung adenocarcinoma by regulating the expression of OPLAH.

Wei Chen, Kai Wang, Renyu Wang, Xufeng Deng, Xiaobing Liu, Peng Dai, Quanxing Liu.

  Lung adenocarcinoma (LUAD) is the most common subtype of lung cancer. Glutamine plays a critical role in the progression of LUAD. However, the function of pyrroline-5-carboxylate reductase 1 (PYCR1) and its regulatory role in glutamine metabolism remain unclear. Transcriptomic and clinical data for LUAD were obtained from The Cancer Genome Atlas (TCGA) and validated using Gene Expression Omnibus (GEO) datasets (GSE19188, GSE13213). Glutamine metabolism-related genes were analyzed for differential expression and prognostic significance. Functional enrichment was performed via gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) analyses. Single-cell RNA-seq data (GSE117570) were processed using Seurat, and cell-cell communication was inferred with CellChat. In vitro, lentiviral overexpression, Western blotting, EdU, CCK-8, and glutamine uptake assays were conducted. An orthotopic xenograft model was established in nude mice to assess tumor growth in vivo. Six glutamine-metabolism-related genes were found significantly overexpressed in LUAD tissues and associated with poor overall survival. Single-cell sequencing revealed predominant PYCR1 expression in malignant cells. Functional assays demonstrated that PYCR1 overexpression enhanced glutamine uptake, proliferation, and inhibited apoptosis in LUAD cells, effects mediated via suppression of the P53 pathway. PYCR1 promoted tumor growth in a xenograft model and was found to transcriptionally upregulate 5-oxoprolinase (OPLAH), which augmented its oncogenic effects. Our findings identify the PYCR1/OPLAH axis as a key driver of LUAD progression via p53 signaling, revealing a promising therapeutic target.

Keywords:  5-oxoprolinase; Glutamine; Lung adenocarcinoma; pyrroline-5-carboxylate reductase 1

DOI:  https://doi.org/10.1007/s10238-026-02181-9
Curr Med Chem. 2026 Jun 10.

Metabolic Reprogramming and Molecular Mechanisms in Renal Cell Carcinoma: Therapeutic Implications and Future Perspectives.

Ferdos Faghihkhorasani, Guodong Zhu.

  Renal cell carcinoma (RCC) is a biologically heterogeneous malignancy with distinct metabolic dependencies that differentiate it from many other solid tumors. Despite recent advances in targeted therapies and immunotherapies, therapeutic resistance and variable clinical responses remain major challenges, underscoring the need for a deeper understanding of RCC-specific metabolic vulnerabilities. Current evidence indicates that metabolic reprogramming is a central driver of RCC progression, involving enhanced glycolysis, glutaminolysis, one-carbon metabolism, altered lipid metabolism, and mitochondrial adaptations. These metabolic shifts are largely regulated by dysregulated oncogenic signaling, constitutive activation of hypoxia-inducible factors (HIFs), and dynamic interactions within the tumor microenvironment. Key metabolic regulators and enzymes, including HIF-2α, glutaminase (GLS), fatty acid synthase (FASN), and methylenetetrahydrofolate dehydrogenase 2 (MTHFD2), have emerged as clinically relevant targets with therapeutic potential. In this review, we synthesized current knowledge on RCC metabolism while highlighting features that distinguish RCC from other malignancies, particularly its HIF-driven metabolic landscape and pronounced microenvironmental influences. Importantly, we extended descriptive metabolism by focusing on clinically actionable pathways, biomarker-driven patient stratification, and rational combination strategies integrating metabolic inhibitors with immunotherapy or targeted agents. We also discussed some emerging methodologies, including metabolic imaging and spatial profiling approaches, to address intratumoral metabolic heterogeneity. Overall, this review emphasizes how leveraging RCC-specific metabolic vulnerabilities can inform precision medicine approaches and improve therapeutic outcomes for patients with RCC.

Keywords:  HIF-2α inhibitors; VHL-HIF axis; glutamine metabolism; lipid metabolism.; tumor microenvironment

DOI:  https://doi.org/10.2174/0109298673461719260530152756
iScience. 2026 Jun 19. 29(6): 116347

Semaglutide targets muscle mitochondria to regulate glutamine metabolism and treat osteoarthritis.

Yuchen Tian, Zhikai Zheng, Yiyang Ma, Fang Ye, Yidan Pang, Zhicheng Zhang, Zhiwei Li, Changqing Zhang, Junjie Gao.

  As a metabolic regulatory drug for lowering body weight, semaglutide has shown additional therapeutic effects in osteoarthritis (OA), but the underlying mechanism requires further investigation. Here, we employed cross-tissue single-cell RNA sequencing analysis and found that semaglutide improved mitochondrial metabolic disorders in muscle tissue under high-fat diet and OA conditions in mice. The results of multi-omics analysis indicated that semaglutide targeted muscle mitochondria to regulate glutamine metabolism during OA. Intramuscular injection of mitochondria from C2C12 cells, pre-stimulated by semaglutide, could alleviate pain symptoms and cartilage damage of OA, which was achieved by inhibiting muscle glutaminase activity and upregulating circulating glutamine concentration. In vitro experiments confirmed the alleviating effect of glutamine produced by C2C12 cells stimulated by semaglutide on chondrocyte inflammation. These findings reveal the mitochondrial regulatory mechanism in the muscle-cartilage axis and provide a perspective for the application of semaglutide in OA treatment.

Keywords:  human metabolism; musculoskeletal medicine; therapeutics

DOI:  https://doi.org/10.1016/j.isci.2026.116347
J Biochem Mol Toxicol. 2026 Jun;40(6): e70924

Posttranscriptional Regulation of Metabolism in Glioblastoma: A Multipathway Review.

Suleiman Ibrahim Mohammad, Asokan Vasudevan, Favian Bayas Morejón, Bobur Abduvoyitov, Tina Saeed Basunduwah, Zahraa Khudhair Abbas Al-Khafaji, Basma Mohammed Abdulhussein, Ahmed Hjazi, Khetam Habeeb Rasool, Majid S Jabir.

  Glioblastoma (GBM) is the most aggressive and lethal form of primary brain tumor. A hallmark of GBM metabolism is the Warburg effect, whereby tumor cells preferentially utilize aerobic glycolysis despite oxygen availability, producing ATP inefficiently but supporting anabolic processes. Concurrently, the pentose phosphate pathway (PPP), amino acid metabolism, lipid biosynthesis, and nucleotide synthesis are rewired to meet the energetic and biosynthetic demands of GBM cells. Recent discoveries underscore the role of microRNAs (miRNAs) as master regulators orchestrating these metabolic rewiring events. Acting posttranscriptionally, miRNAs target key transporters, enzymes, and signaling molecules involved in glycolysis, glutaminolysis, lipid biosynthesis, and oxidative metabolism. This review explores how miRNA networks modulate metabolic plasticity in GBM. Specific miRNAs, such as miR-153, miR-451, miR-940, and miR-200b, suppress glutamine metabolism, regulate glucose transporters (e.g., GLUT1/3), inhibit lactate dehydrogenase, and disrupt mitochondrial folate metabolism. Others, such as miR-29 and miR-183, control lipid and nucleotide metabolism via the SREBP1 and IDH2 pathways. Furthermore, regulatory interactions among miRNAs, long non-coding RNAs (lncRNAs), and circular RNAs (circRNAs), such as the XIST/miR-126 or circ-CREBBP/miR-375 axes, create complex feedback loops that fine-tune metabolic pathways and enhance tumor survival under stress. We also discuss therapeutic strategies targeting these miRNA-metabolism circuits, including nanoparticle delivery, dietary restriction, and combination therapies that re-sensitize tumors to temozolomide and radiation. Understanding and therapeutically exploiting these networks presents a powerful approach to overcoming GBM's metabolic resilience, thereby opening new avenues for precision oncology.

Keywords:  glioblastoma; glycolysis; metabolic reprogramming; microRNAs; therapeutics

DOI:  https://doi.org/10.1002/jbt.70924
Am J Physiol Endocrinol Metab. 2026 Jun 15.

Glutamine metabolism shapes the phenotypic adaptation of uterine dendritic cells in response to early allogeneic pregnancy.

Songchen Cai, Agnes Wieczorek, Christopher Urbschat, Maria Emilia Solano, Yong Zeng, Petra Clara Arck, Lianghui Diao, Kristin Thiele.

  The impact of metabolic reprogramming on immune cell functions is increasingly recognized. However, it remains largely unexplored in terms of immune cells adaptation during reproduction. Dendritic cells (DC) are crucial for establishing and maintaining pregnancy by orchestrating maternal immune adaptation essential for embryo implantation and decidualization. Here, we characterized the phenotypic and metabolic characteristics of DCs during early pregnancy in an allogeneic mouse model and in response to the specific deletion of hormonal receptors on DCs. Frequency of uterine CD11c+ DCs on gestational day (gd) 7.5 remained equal to that of non-pregnant mice. However, we observed a functional shift from cDC1 to cDC2 in pregnant mice. In parallel, a metabolic switch in uterine DCs was identified by upregulation of genes representing fatty acid synthesis (Fasn, Acaca), fatty acid oxidation (Cpt1a), and glutamine-related metabolic pathways (Got2). The cell-specific deletion of the glucocorticoid receptor in DCs reduced their MHCII expression, accompanied by a reduction in Got2 expression. Glutamine deprivation in vitro dramatically reduced the absolute number of cultured bone marrow cells and the frequency of cDC1s while simultaneously increasing the frequency of cDC2s. Collectively, these findings establish glutamine metabolism as a key driver of DC adaptation during early pregnancy, revealing novel metabolic-immunological crosstalk at the maternal-fetal interface.

Keywords:  Allogeneic pregnancy; Dendritic cells; Glutamine; Immunometabolism; Pregnancy hormones

DOI:  https://doi.org/10.1152/ajpendo.00520.2025
PeerJ. 2026 ;14 e21400

The metabolic escape: how tumor metabolic reprogramming drives drug resistance.

Yan Chen, Zixu Wu, Wenzhe Si, Xujun Liu.

  In the last few years, metabolic reprogramming has been recognized as a fundamental characteristic of cancer, and is also acknowledged as a crucial cause to drug resistance, which consistently acts as a significant barrier in cancer treatment by allowing tumor cells to adapt and escape various therapies. This review gives a systematically investigation of how metabolic reprogramming contributes to drug resistance in cancer, including aerobic glycolysis (also known as the Warburg effect), lactate metabolism, glutamine addiction, lipid synthesis reprogramming, mitochondrial and ion metabolic changes. Furthermore, by clarifying the mechanisms behind these reprogrammed metabolic pathways, we explain how these changes lead to drug resistance and highlight potential molecular targets for therapeutic intervention. Additionally, we discuss emerging strategies aimed at exploiting these metabolic vulnerabilities, offering new insights for overcoming drug resistance in cancer. By integrating recent discoveries in this field, we present a unified perspective on targeting metabolic vulnerabilities to overcome drug resistance, which is an urgent need in precision oncology, and timely and concise insights for cancer biologists and researchers in the field of exploring the metabolic mechanisms of drug resistance. We hope this review will provide valuable insights for molecular tumor biologists seeking to elucidate the molecular roles of tumor metabolic reprogramming and drug resistance in cancer.

Keywords:  Cancer; Cancer therapeutics; Drug resistance; Metabolic reprogramming; Tumor microenvironment

DOI:  https://doi.org/10.7717/peerj.21400
Front Immunol. 2026 ;17 1815245

Metabolic reprogramming of glutamine is associated with M2 macrophage polarization in allergic rhinitis.

Shang Yan, Kai Xue, Yinan Wang, Desheng Jia, Yongchao Chen, Xueyou Lai, Chunrui Zhao, Guowei Chen, Yang Xu, Xiaoxu Wang, Hongguang Pan, Nan Zeng.

   Introduction: Allergic rhinitis (AR) is classically regarded as a type 2 immune-driven disease, yet its chronicity and heterogeneity suggest that additional regulatory layers shape the local immune microenvironment. Although immune cell function is closely linked to metabolic state, how specific metabolic cues are integrated into immune signaling in AR remains unclear.
Methods: We combined human nasal mucosal metabolomic analysis, murine models, single-cell transcriptomics, bulk RNA sequencing, and protein assays to investigate the role of glutamine in AR pathophysiology.
Results: We identified elevated glutamine levels in AR nasal mucosa and found that dietary glutamine supplementation was associated with altered nasal behavioral responses in experimental AR. Single-cell profiling revealed changes in immune cell composition, with macrophages displaying a shift toward an M2-like transcriptional state under high-glutamine conditions. Transcriptomic and pathway analyses positioned fibroblast growth factor receptor 1 (FGFR1) within differentially enriched signaling networks, and its expression increased under high-glutamine conditions. Protein-level assays further showed that aminoacylation-associated signals on FGFR1 varied with glutamine availability, together with coordinated changes in YARS and SIRT1.
Discussion: These hypothesis-generating findings support an associative model in which metabolic alterations in AR are linked to non-canonical modification of FGFR1 and macrophage transcriptional polarization, suggesting a potential immunometabolic layer regulating the nasal mucosal microenvironment.

Keywords:  fibroblast growth factor receptor 1; glutamine; immunometabolism; macrophage polarization; metabolomics; single-cell RNA sequencing

DOI:  https://doi.org/10.3389/fimmu.2026.1815245
iScience. 2026 Jun 19. 29(6): 116253

The competition of amino acid among tumors, tumor-associated macrophages and T cells based on metabolic reprogramming.

Chenyang Xianyu, Yuxiao Song, Ci Zhao, Qian Min, Zhixu Wang, Mingbo Zhang, Bicheng Zhang.

  Amino acids are important nutrients in the process of tumor proliferation. Dysregulated amino acid metabolism profoundly influences tumor growth and immune cell function. Within the tumor microenvironment (TME), metabolic reprogramming of amino acids modulates the polarization of tumor-associated macrophages (TAMs) and the differentiation of T cells, processes intimately linked to tumor immune evasion. Meanwhile, metabolic reprogramming leads to amino acid competition between tumor cells and immune cells, particularly TAMs and T cells. To meet their own amino acid needs, tumors carry out a series of optimized metabolic strategies by expressing specific enzymes, cytokines, and amino acid transporters, and so forth promoting the formation of an immunosuppressive microenvironment and hindering anti-tumor immunity. Notably, this metabolic competition may exhibit spatial heterogeneity and temporal dynamics. Given the central role of amino acid metabolism in tumor progression and immune evasion, targeting key metabolic pathways represents a promising therapeutic strategy for cancer treatment.

Keywords:  Cancer; Cancer systems biology; Human metabolism; Immune response

DOI:  https://doi.org/10.1016/j.isci.2026.116253
Cell Rep Med. 2026 Jun 16. pii: S2666-3791(26)00262-4. [Epub ahead of print]7(6): 102845

Ketogenic diet as a metabolic vehicle enhancing the therapeutic efficacy of mebendazole and devimistat in juvenile syngeneic high-grade glioma.

Purna Mukherjee, Jack Maurer, Sylwia A Stopka, Bennett Greenwood, Juan J Aristizabal-Henao, Srada Karmacharya, Alexandra Chimento, Nathan Ta, Derek C Lee, Tomas Duraj, Roderick T Bronson, Michael A Kiebish, Thomas N Seyfried.

  Invasive pediatric high-grade gliomas (HGGs) are associated with poor clinical outcomes, and current therapies often cause significant long-term toxicities. This study investigates the influence of nutritional ketosis on the therapeutic efficacy of mebendazole (MBZ) and devimistat (CPI-613) in invasive VM-M3 and non-invasive CT-2A glioblastoma models in juvenile syngeneic mice. Both drugs were also evaluated in the human pediatric glioma cell line SF-188. Mesenchymal-origin VM-M3 tumors exhibited extensive invasion throughout the brain and spinal cord, whereas neural stem cell-derived CT-2A and VM-NM1 tumors did not show distal spread. The greatest reductions in tumor invasion and progression, together with prolonged survival, occurred when drug treatment was combined with a ketogenic diet (KD). MBZ inhibited glycolysis and glutaminolysis in VM-M3 cells and reduced proliferation and viability of SF-188 cells. KD-enabled combination therapy allowed lower drug dosing, reduced toxicity, and improved survival, supporting further investigation of metabolically informed diet-drug strategies for pediatric gliomas.

Keywords:  childhood brain cancer; glucose; glutamine; glutaminolysis; glycolysis; ketones; metabolic therapy; press-pulse; spinal cord

DOI:  https://doi.org/10.1016/j.xcrm.2026.102845
J Am Chem Soc. 2026 Jun 15.

Glutamine Tautomerization Drives RhoGAP-Aided GTP Hydrolysis in Small Rho GTPases.

Angela Parise, Riccardo Rozza, Karolina Mitusinska, Jana Aupič, Alessandra Magistrato.

Rho GTPases promote GTP hydrolysis aided by specific GTPase-activating proteins (GAPs). By alternating between an active GTP-bound and an inactive GDP-bound state, Rho GTPases function as molecular switches regulating cytoskeletal dynamics and cell motility. Despite their biological relevance, the detailed molecular mechanism underlying Rho GTPases catalysis remains contentious. Here, using classical and hybrid quantum-classical molecular dynamics, we resolve the mechanism of GTP hydrolysis in the RhoGAP-RhoA complex. We reveal that GTP hydrolysis proceeds through a dissociative nucleophilic substitution mechanism, driven by an amide → imide tautomerization of Gln63, which aids in delivering a proton from the nucleophilic water to the leaving phosphate group. The Gln63 imide tautomer also loosens RhoGAP-RhoA interfacial contacts, allowing solvent molecules to enter and drive a water-mediated reverse tautomerization of Gln63 that restores the catalytically competent configuration of the RhoA active site. Conservation of key interface residues across Rho/Rho GAP family members suggests that this mechanism may be shared by most Rho GTPases.

DOI: https://doi.org/10.1021/jacs.6c06153
Arch Med Res. 2026 Jun 13. pii: S0188-4409(26)00079-2. [Epub ahead of print]57(6): 103456

Protective Effects of Glycyl-Glutamine Dipeptide on In Vitro Brain Ischemia Reperfusion Model and the Role of Sigma-1 Receptors.

Gülce Sevdar Çeçen, Sümeyye Keskin, Mustafa Sertaç Yılmaz, Rıfat Levent Büyükuysal, Mine Sibel Gürün, Pınar Ellergezen, Sinan Çavun.

   BACKGROUND: Cerebral ischemia-reperfusion injury is characterized by oxidative stress, inflammation, and apoptotic cell death, which results in tissue damage. Although the Glycyl-Glutamine (Gly-Gln) dipeptide has primarily been investigated as a glutamine donor, its direct pharmacological effects and underlying mechanisms remain poorly defined.
METHODS: In this study, the effects of Gly-Gln on tissue viability and biochemical parameters were investigated using an in vitro model of cerebral ischemia-reperfusion injury. Dose-response analysis was performed using 2,3,5-triphenyltetrazolium chloride (TTC) staining to determine the minimum effective concentration of Gly-Gln. To assess whether the observed effects were specific to the dipeptide, equimolar concentrations of its degradation products, glycine and glutamine, were also evaluated. Oxidative stress and apoptotic markers were analyzed, and the potential involvement of sigma receptor ligands was examined.
RESULTS: Gly-Gln improved tissue viability in a dose-dependent manner, with 1,000 µM identified as the minimum effective concentration. Equimolar administration of glycine or glutamine did not produce comparable protective effects, indicating that the observed actions were specific to the intact dipeptide. Ischemia-reperfusion significantly increased lipid peroxidation and disrupted antioxidant balance, as evidenced by elevated malondialdehyde levels and reduced glutathione content. Although Gly-Gln alone did not significantly attenuate lipid peroxidation at the tested concentration, modulation of sigma-1 receptors markedly reduced oxidative damage. Apoptotic activity was increased following ischemia-reperfusion, whereas sigma receptor agonist treatment decreased the expression of apoptotic markers.
CONCLUSION: In conclusion, Gly-Gln exerts direct, dose-dependent protective effects on tissue viability in cerebral ischemia-reperfusion injury, independently of its degradation products, and sigma-1 receptors may contribute to the underlying mechanisms.

Keywords:  Gly-Gln; Ischemia-reperfusion; Neuroprotection; Sigma-1 receptors

DOI:  https://doi.org/10.1016/j.arcmed.2026.103456
Toxicol Appl Pharmacol. 2026 Jun 16. pii: S0041-008X(26)00215-2. [Epub ahead of print]514 117919

Growth-mode-independent glutamine/glutamate/aspartate metabolic shift and urea cycle remodeling driven by FOXA1 in benzo[a]pyrene-transformed human bronchial epithelial cells.

Yujin Fu, Yanmin Qi, Yunkun Xing, Yitong Lin, Zhiyu Liu, Juanling Fu, Biyun Yao, Peng Zhao.

  To characterize the amino acid metabolic remodeling in THBEc1 cells malignantly transformed by benzo[a]pyrene (BaP) relative to parental human bronchial epithelial HBE cells, and to delineate the role of FOXA1 in this remodeling, the amino acid metabolic phenotypes and gene expression patterns of HBE, THBEc1, THBEc1-ctrl (knockout control), and THBEc1-FOXA1 KO (FOXA1 knockout) cells were profiled under 2D, 3D spheroid, and in vivo conditions. Differential metabolic amino acids across these growth modes were identified. THBEc1 cells exhibited a glutamine/glutamate/aspartate metabolic shift and urea cycle remodeling that were conserved across all growth modes, representing key metabolic features of BaP-induced malignant transformation. FOXA1 knockout partially reversed these metabolic alterations across growth modes. Mechanistically, FOXA1 transcriptionally regulated GLUL, SLC6A14, CPS1, and SLC7A2 in a growth-mode-independent manner. These findings establish FOXA1 as a key mediator of the glutamine/glutamate/aspartate metabolic shift and urea cycle remodeling during BaP-induced malignant transformation, with GLUL, SLC6A14, CPS1, and SLC7A2 as critical downstream targets. This work advances the understanding of amino acid metabolic reprogramming in BaP-induced lung carcinogenesis.

Keywords:  3D spheroid culture; FOXA1; Gln/Glu/Asp metabolic shift; In vivo xenograft model; THBEc1 cells; Urea cycle remodeling

DOI:  https://doi.org/10.1016/j.taap.2026.117919
J Pharm Biomed Anal. 2026 Jun 13. pii: S0731-7085(26)00280-3. [Epub ahead of print]280 117612

Development of new liquid chromatography-tandem mass spectrometry methods for the quantification of seven metabolites in three related breast cancer cell lines.

Marziyeh Ghanbarian, Deborah Michel, Maryam Alyari, Amir Khajavinia, Oleg Y Dmitriev, Anas El-Aneed.

  Metabolites of the tricarboxylic acid (TCA) cycle play crucial roles in cancer biology, and their accurate quantification is essential for understanding energy metabolism, signaling dynamics, and identifying metabolic vulnerabilities in cancer cells. However, traditional liquid chromatograph-tandem mass spectrometry (LC-MS/MS) methods for these polar metabolites often encounter challenges, such as limited retention on reversed-phase columns and ion suppression. This study developed and validated two LC-MS/MS methods for the accurate quantification of seven key TCA cycle metabolites in MDA-MB-231, M67-2 (MEMO1 knockdown), and M67-9 (MEMO1 knockout) breast cancer cell lines. For five metabolites, namely citrate (CA), L-malate (MA), fumarate (FA), α-ketoglutarate (AKG), and glutamate (GA), an isotope-coded derivatization approach utilizing 12C/13C-labeled dimethylaminophenacyl (DmPA) bromide was employed to develop a targeted high-performance liquid chromatography (HPLC)-MS/MS method. Inefficient DmPA derivatization in aqueous matrices was addressed by optimizing sample preparation in non-aqueous conditions, and the presence of multiple peaks of AKG was resolved by selecting triethanolamine (TEOA) as the reaction base to improve specificity. Conversely, due to persistent interferences with DmPA derivatization, pyruvic acid (PA) and succinic acid (SA) were quantified using another novel hydrophilic interaction liquid chromatography (HILIC)-MS/MS method in their native underivatized forms. Both methods were validated according to regulatory bodies, ensuring linearity, accuracy, precision, selectivity, and stability. The methods ensured the utilization of two multiple reaction monitoring (MRM) transitions to enhance specificity. The validation approach was adjusted to fit tissue culture studies. The validated methods were successfully used to measure the TCA metabolites in tested cell lines, providing valuable tools for investigating metabolic dynamics in cancer research.

Keywords:  Cancer cells; DmPA derivatization; HILIC-MS/MS; HPLC-MS/MS; LC-MS/MS; TCA cycle metabolites; Targeted metabolomics

DOI:  https://doi.org/10.1016/j.jpba.2026.117612
Bioanalysis. 2026 Jun 17. 1-13

Targeted LC-MS/MS profiling of serum amino acids reveals type 2 diabetes-specific alterations in advanced gastric cancer.

Mehmet Emrah Yaman, Omer Faruk Kocak, Omer Topdagi, Yucel Kadioglu.

   BACKGROUND: Altered amino acid metabolism is a feature of gastric cancer. Type 2 diabetes (T2D), a prevalent metabolic comorbidity, may affect circulating amino acid profiles; this study aimed to quantify its impact.
RESEARCH DESIGN AND METHODS: A validated targeted LC-MS/MS assay profiled 22 serum amino acids in 156 participants, including gastric cancer patients with and without type 2 diabetes and controls.
RESULTS: Adjusted models showed that many observed differences were attributable to type 2 diabetes and body mass index. Type 2 diabetes was associated with lower ornithine (β = -1.16; q = 0.0019) and interacted with gastric cancer for arginine (β_int = -1.10; q = 0.0044) and phenylalanine (β_int = +1.05; q = 0.0112). After adjustment, only valine, taurine, and total amino acids remained lower in gastric cancer. Body mass index was inversely associated with glutamine and positively associated with glutamate.
CONCLUSIONS: Covariate-adjusted analysis distinguishes cancer-related from comorbidity-driven alterations and identifies type 2 diabetes as a key modifier of amino acid metabolism. Metabolic covariates should be considered when interpreting amino acid alterations in advanced gastric cancer biomarker studies. However, the findings should be interpreted in light of the predominantly advanced-stage gastric cancer cohort and require validation in larger prospective studies.

Keywords:  Targeted LC–MS/MS; amino acid profiling; covariate-adjusted analysis; gastric cancer; type 2 diabetes

DOI:  https://doi.org/10.1080/17576180.2026.2684669
J Proteome Res. 2026 Jun 18.

Evaluation of FASP for Mass Spectrometry-Based Untargeted Metabolomics Analysis of Urine Samples.

Muath Khairi Mousa, Luis B Carvalho, Alexander D Giddey, André Figueiredo, Hamza Al-Hroub, Mohammad H Semreen, Mohammed Uddin, Hugo M Santos, Nelson C Soares.

  Filter-Aided Sample Preparation (FASP) is a well-established method in proteomics, yet its potential for the parallel recovery of metabolites remains largely unexplored. Herein, we evaluate the performance of FASP as a straightforward workflow for the simultaneous isolation of protein and corresponding metabolite fractions from a single urine sample. The FASP-based LC-MS/MS approach for both proteomics and metabolomics analysis identified 3,163 nonredundant peptides corresponding to 957 unique protein groups. The metabolomic profile comparison of three urine fractions, specifically FASP-concentrated, FASP flow-through, and raw samples, resulted in the identification of 176 common metabolites. Next, as a proof-of-concept, the FASP protocol was applied to compare the metabolomic profiles of clinical urine samples from healthy individuals (n = 13) and patients with Ta bladder cancer (n = 12). The metabolomic modulation was consistent with previously reported findings, highlighting perturbations in phenylacetate, purine, and tryptophan metabolism, as reflected by changes in metabolites such as adenosine monophosphate (AMP), phenylacetic acid, glutamine, cytosine, and l-tryptophan. FASP protocol can be effectively adapted for the concurrent profiling of both proteomic and metabolomic fractions from urine samples. Thus, FASP-based workflow represents a viable alternative for single-step sample preparation, facilitating subsequent quantitative multiomics data integration.

Keywords:  Bladder cancer; FASP; Metabolomics; Multiomics; Proteomics; Sample preparation; Urine

DOI:  https://doi.org/10.1021/acs.jproteome.6c00173