Nuclear magnetic resonance metabolic fingerprint of bevacizumab in mutant IDH1 glioma cells

Abstract

BackgroundMalignant gliomas are rapidly growing tumours that extensively invade the brain and have bad prognosis. Our study was performed to assess the metabolic effects of bevacizumab on the glioma cells carrying the IDH1 mutation, a mutation, associated with better prognosis and treatment outcome. Bevacizumab is known to inhibit tumour growth by neutralizing the biological activity of vascular endothelial growth factor (VEGF). However, the direct effects of bevacizumab on tumour cells metabolism remain poorly known.Materials and methodsThe immunoassay and MTT assay were used to assess the concentration of secreted VEGF and cell viability after bevacizumab exposure. Metabolomic studies on cells were performed using high resolution magic angle spinning spectroscopy (HRMAS).ResultsmIDH1-U87 cells secreted VEGF (13 ng/mL). Regardless, bevacizumab had no cytotoxic effect, even after a 72h exposure and with doses as high as 1 mg/mL. Yet, HRMAS analysis showed a significant effect of bevacizumab (0.1 mg/mL) on the metabolic phenotype of mIDH1-U87 cells with elevation of 2-hydroxyglutarate and changes in glutamine group metabolites (alanine, glutamate, glycine) and lipids (polyunsaturated fatty acids [PUFA], glycerophosphocholine, and phosphocholine).ConclusionsIn mIDH1-U87 cells, changes in glutamine group metabolites and lipids were identified as metabolic markers of bevacizumab treatment. These data support the possibility of a functional tricarboxylic acid cycle that runs in reductive manner, as a probable mechanism of action of bevacizumab in IDH1 mutated gliomas and propose a new target pathway for effective treatment of malignant gliomas.

Metabolomic and Lipidomic Profiling of Gliomas—A New Direction in Personalized Therapies

“…In addition, Fack et al [ 48 ] also reported the down-regulation of the γ-glutamyl cycle and a reduction in all associated metabolites, including cysteine, glutamate, glycine, and glutathione [ 48 ]. Most of the above-mentioned findings were confirmed in mutant IDH1 glioma cells by Mesti et al [ 47 ], who found significant changes not only in the metabolites from the glutamine group (e.g., glutamate, alanine, and glycine) but also in the lipids. In this work, Mesti et al observed modified levels of polyunsaturated fatty acids (PUFA), glycerophosphorylcholine (GPC), and phosphatidylcholine (PC) following bevacizumab treatment and considered these changes early markers of metabolic alterations due to this treatment [ 47 ].…”

“…Most of the above-mentioned findings were confirmed in mutant IDH1 glioma cells by Mesti et al [ 47 ], who found significant changes not only in the metabolites from the glutamine group (e.g., glutamate, alanine, and glycine) but also in the lipids. In this work, Mesti et al observed modified levels of polyunsaturated fatty acids (PUFA), glycerophosphorylcholine (GPC), and phosphatidylcholine (PC) following bevacizumab treatment and considered these changes early markers of metabolic alterations due to this treatment [ 47 ]. The identification of biochemical pathways affected by bevacizumab enables the proposition of potentially effective combination therapies.…”

Two-Week Aflibercept or Erlotinib Administration Does Not Induce Changes in Intestinal Morphology in Male Sprague–Dawley Rats But Aflibercept Affects Serum and Urine Metabolic Profiles

“…Although the authors attributed this effect to temrolimus, our results suggest that VEGF inhibition alone can increase serum lipid concentrations. In vitro studies with different glioma cell lines have also demonstrated increased levels of fatty acids and changes in choline metabolism after pharmacological treatment with VEGF receptor 2 inhibitor or with bevacizumab [30], [31]. Overall, similar metabolic alterations have also been associated with increased apoptosis in several cell lines [30], [31], [32], [33], [34].…”

“…In vitro studies with different glioma cell lines have also demonstrated increased levels of fatty acids and changes in choline metabolism after pharmacological treatment with VEGF receptor 2 inhibitor or with bevacizumab [30], [31]. Overall, similar metabolic alterations have also been associated with increased apoptosis in several cell lines [30], [31], [32], [33], [34]. Although aflibercept does not appear to overtly induce apoptosis in cell models [35], [36], [37], [38], the metabolic shifts observed in our study could reflect aflibercept-mediated in vivo tissue hypoxia and oxidative stress that drive apoptotic processes.…”

High‐resolution magic angle spinning NMR for intact biological specimen analysis: Initial discovery, recent developments, and future directions

“…Most cell-line studies are conducted in the domain of defining metabolomic profiles or testing treatment efficacy for various malignant and nonmalignant diseases. 40,[136][137][138][139][140][141][142][143][144][145][146][147][148] In addition, a number of studies have used HRMAS NMR to investigate cell structures and real-time metabolic reactions.…”