Table 2.

Summary of studies employing metabolomics in GB research.

Application	Aim	Tissue Type /Cell Line	Technique	Key Metabolites	Key Metabolic Pathways	Principle Insights	Reference
Biochemical characterization	Differentiate glioblastoma subtypes; define infiltrative tumor boundaries; potential utility in evaluating treatment effects.	Tumor and peritumoral GB tissue	MALDI-TOF-MSI (matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry imaging)	Antioxidants Fatty acids Purine and pyrimidine metabolites Reduced N-acetylaspartate abundance, etc.	Purine and pyrimidine metabolism, arachidonic acid synthesis, TCA cycle.	Metabolic information obtained could enhance and customize therapy methods The study underlines MSI’s appropriateness for GB research	[86]
Biochemical characterization	Create a xenograft for GB therapeutic testing; investigate the link between treatment efficacy and tumor metabolism.	Glioblastoma xenograft tissue	MALDI –Fourier transform ion cyclotron resonance (FT-ICR)-MSI	Heme ATP Acylcarnitine	Glycolysis, fatty acid metabolism, antioxidant, and anti-apoptotic functions.	Cells in the tumor’s core and edge experience distinct fatty acid metabolism, leading to different chemical microenvironments within the tumor.This can impact medication distribution via changes in tissue drug affinity or transport and is an essential consideration for therapeutic options in the treatment of GB.	[87]
Pharmacometabolomic approach	To investigate, for the first time, the influence of glabrescione B (GlaB), a known Hedgehog (Hh) pathway inhibitor, on glioma cell proliferation and metabolism in in vivo and in vitro models.	Murine glioma cells (GL261)	1H-NMR, HPLC–MS	Lactate Glycine Tyrosine Phenylalanine Histidine Alanine Leucine Isoleucine Valine	Glycolytic metabolism.	The endo- and exo-metabolomes of GlaB-treated and untreated cells exhibited changes in metabolite levels over time. GlaB, a direct inhibitor of the transcription factor Gli1, suppresses glioma cell proliferation while exacerbating the Warburg effect.	[88]
Comparative biomarker discovery	Because altered tumor metabolism is one of the hallmarks of cancer, the aim was to explore if the rate-limiting enzyme argininosuccinate synthetase (ASS1) positive and negative GB cell lines had distinct metabolic profiles that may allow for non-invasive diagnosis and reveal new treatment prospects.	GAMG LN229 SNB19 T98G U118 U87 Normal Human Astrocytes (NHA)	One and two-dimensional gas chromatography-time-of-flight mass spectrometry (1D/2D GC-TOFMS), LC-TOFMS.	Mannose Galactose Glucose Pyruvic acid Citrate α-ketoglutaric acid	Not detected.	The metabolome contains systematic information distinguishing between ASS1 positive and negative GB cell lines. There is a possibility of identifying metabolite biomarkers for the non-invasive detection from these subtypes, as well as the identification of novel treatment targets.	[89]
Pharmacometabolomic approach	The goal of this trial was to see if carboplatin worked in tandem with the mTOR complex 1 inhibitor (everolimus) in pediatric low-grade glioma (pLGG).	pLGG cell line BT66 JHH-NF1-PA1 Res259 Res18	LC–MS	Glutathione Glutamine Glutamate	Comparable pathways were discovered in patient-derived xenograft in mice.	The combination of everolimus and carboplatin works synergistically in pLGG. The study confirms a novel therapy regimen that may be promptly pushed into pediatric phase I/II clinical trials. This work presents a justification for novel mTORC1-based inhibitor therapy combinations in brain malignancies.	[90]
Pharmacometabolomic approach	To investigate the effect of glutaminase (GLS) inhibition on GSCs, which have been implicated in the development of medication resistance and tumor recurrence.	1H-NMR JHH520 GBM1 268, 407, 23, 233, 349 SF188 NCH644	1H-NMR	Alanine Aspartate Glutamine Glutamate Glycine Glutathione Lactate Myo-inositol Succinate Tricarboxylic acid Total choline	Not detected	The findings demonstrate the use of in vitro pharmaco-metabolomics for therapeutic effectiveness evaluation and compound risk assessment. It emphasizes the importance of GLS as a druggable and prospective therapeutic target in our desire to enhance the management of GB medication resistance and tumor relapse by focusing on GSCs subpopulation.	[91]
Developmental therapeutics	To develop standardized pediatric high-grade gliomas (pHGGs) models for drug testing and to generate an exact physiological brain environment in vitro.	Primary glioblastoma	NMR	Acetate Alanine Beta-glucose Choline Creatine Glutamate Glycerophosphocholine Glycine Lactate Myo-inositol N Acetylaspartate, Serine Taurine Valine	Some pathways were altered in the 2D/3D cell cultures pathways in patient tumor relapse.	A hypoxic environment helps to preserve the original patient tumor metabolism and characteristics. The multi-step effort may be regarded as a standard for developing therapeutically relevant models.	[92]
Developmental therapeutics	Researchers hypothesized that the branched-chain α-ketoacids (BCKA) depletion is caused by the (R) enantiomer of 2-hydroxyglutarate((R)-2HG)’s direct, competitive suppression of branched chain amino acids transaminases (BCAT) activity.	GSC lines: TS603, TS516, MGG152, TS676, BT054 BT260NHA HT1080 HOG IDH1 R132H mutant IDH2 R172K mutant HCT116 HEK293T NCI-H82	GC-MS, hybrid triple quadrupole mass spectrometer, Hydrophilic interaction liquid chromatography(HILIC)	Alpha-Keto-beta-methylvalerate Alpha-Ketoisocaproate Glutamate 2-hydroxyglutarate Leucine Valine isoleucine	Increased BCAT activity in vitro and in vivo.	BCAT suppression produces metabolic vulnerabilities that can be leveraged therapeutically to sensitize IDH mutant gliomas. ((R)-2HG is overproduced in IDH mutant GBs). Gliomas with IDH mutations are more sensitive to radiation when combined with glutaminase inhibition, suggesting a novel way to treating these tumors.	[93]
Developmental therapeutics	To assess the effect of paclitaxel and/or etoposide on the molecular changes in GB cells	U87 U373	Ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS)	Nutriacholic acid L-phenylalanine L-arginine Guanosine ADP Hypoxanthine guanine	Urea and citric acid cycles Metabolism of polyamines and amino acids	The results can be used to map the anticancer activity of paclitaxel and/or etoposide within the cancer cells under investigation.	[94]
Biomarker discovery	To use NMR spectroscopy to characterize the metabolome of tiny EVs or exosomes produced from distinct GB cells and compare them to the metabolic profile of their parental cells.	NHA U118 LN-18 A172	1H-NMR	Asparagine Acetone Carnitine Ethanol Formate Glycerol malate GSSG GSH GABA G6P Glucose Isoleucine Taurocholic acid Niacinamide lactate 5-oxoproline Citrate Proline succinate Homoserine Glycine	Not detected	The findings revealed a distinct divergence in the metabolic profiles of GB cells, EVs, and medium. The findings are reviewed in relation to new GB diagnostics and therapy monitoring.	[95]
Biomarker discovery	To describe a transcriptional adaption regulatory system that is influenced by environmental factors.	Primary GB	1H-NMR	Alpha-ketoglutarate Arginine Caproic acid Choline Dodecanoic acid Fructose Fumarate Glyceraldehyde Glutathione Glycine Guaiacol Glucose-6-phosphateLysine Succinic acid Serine Selenomethionine	Comparable metabolic environment spatial disparities	A multi-regional examination of a glioblastoma patient biopsy indicated complex metabolic landscape with varied degrees of hypoxia and creatine enrichment. In creatine-enriched settings, the glycine cleavage system, and hypoxia-inducible factor-1α (HIF1A) destabilization were changed, resulting in transcriptional adaptability.	[96]
Biomarker discovery	To test the hypothesis that GB plasma metabolite profiles may predict clinical outcomes.	Primary and recurrent glioblastoma	LC- triple quadrupole- MS	Arginine Kynurenate Methionine	N/A	The study discovered numerous plasma metabolites that are predictive in glioblastoma patients.	[97]
Biomarker discovery	To investigate the effects of a survivin inhibitor (pro-apoptotic effect) on the metabolome of primary GSCs to look for treatment response signals.	GSCs cultures established from IDH-wildtype GB tumor	NMR spectroscopy	Citrate Lactate	N/A	In comparison to spectrometry-based proteomics, the metabolomics technique used generated alternative biomarker possibilities, highlighting the benefits of complementary approaches. Citrate and lactate are magnetic resonance spectroscopy (MRS) -visible, therefore, these first findings provide the groundwork for further research into in vivo MRS of brain malignancies. NMR metabolomics, when combined, is a technique for tackling glioblastoma.	[98]