1H-NMR metabolomics reveals the Glabrescione B exacerbation of glycolytic metabolism beside the cell growth inhibitory effect in glioma

Abstract

Background Glioma is the most common and primary brain tumors in adults. Despite the available multimodal therapies, glioma patients appear to have a poor prognosis. The Hedgehog (Hh) signaling is involved in tumorigenesis and emerged as a promising target for brain tumors. Glabrescione B (GlaB) has been recently identified as the first direct inhibitor of Gli1, the downstream effector of the pathway. Methods We established the overexpression of Gli1 in murine glioma cells (GL261) and GlaB effect on cell viability. We used 1 H-nuclear magnetic resonance (NMR) metabolomic approach to obtain informative metabolic snapshots of GL261 cells acquired at different time points during GlaB treatment. The activation of AMP activated protein Kinase (AMPK) induced by GlaB was established by western blot. After the orthotopic GL261 cells injection in the right striatum of C57BL6 mice and the intranasal (IN) GlaB/mPEG 5kDa -Cholane treatment, the tumor growth was evaluated. The High Performance Liquid Chromatography (HPLC) combined with Mass Spectrometry (MS) was used to quantify GlaB in brain extracts of treated mice. Results We found that GlaB affected the growth of murine glioma cells both in vitro and in vivo animal model. Using an untargeted 1 H-NMR metabolomic approach, we found that GlaB stimulated the glycolytic metabolism in glioma, increasing lactate production. The high glycolytic rate could in part support the cytotoxic effects of GlaB, since the simultaneous blockade of lactate efflux with α-cyano-4-hydroxycinnamic acid (ACCA) affected glioma cell growth . According to the metabolomic data, we found that GlaB increased the phosphorylation of AMPK, a cellular energy sensor involved in the anabolic-to-catabolic transition. Conclusions Our results indicate that GlaB inhibits glioma cell growth and exacerbates Warburg effect, increasing lactate production. In addition, the simultaneous blockade of Gli1 and lactate efflux amplifies the anti-tumor effect in vivo, providing new potential therapeutic strategy for this brain tumor. Graphical Abstract Electronic supplementary material The online version of this article (10.1186/s12964-019-0421-8) contains supplementary material, which is available to authorized users.

Diauxie and co-utilization of carbon sources can coexist during bacterial growth in nutritionally complex environments

“…Spectral acquisition and processing were performed according to procedures developed at CERM 54,[70][71][72][73][74] . All the spectra were recorded using a Bruker 600 MHz spectrometer (Bruker BioSpin) operating at 600.13 MHz proton Larmor frequency and equipped with a 5 mm PATXI 1 H-13 C-15 N and 2 H-decoupling probe including a z axis gradient coil, an automatic tuning matching and an automatic and refrigerate sample changer (SampleJet).…”

Diauxie and co-utilization are not exclusive during growth in nutritionally complex environments

“…Samples were prepared in 5.00 mm NMR tubes by mixing 60 μL of a potassium phosphate buffer (1.5 M K2HPO4, 100% (v/v) 2 H2O, 10 mM sodium trimethylsilyl [2,2,3,3− 2 H4]propionate (TMSP), pH 7.4) and 540 μL of sample. Spectral acquisition and processing were performed according to procedures developed at CERM [47][48][49][50][51][52] . All the spectra were recorded using a Bruker 600 MHz spectrometer (Bruker BioSpin) operating at 600.13 MHz proton Larmor frequency and equipped with a 5 mm PATXI 1 H-13 C-15 N and 2 H-decoupling probe including a z axis gradient coil, an automatic tuning-matching (ATM) and an automatic and refrigerate sample changer (SampleJet).…”

The SHH/GLI signaling pathway: a therapeutic target for medulloblastoma

“…In adult, the activity of the HH pathway is significantly reduced, except for tissue maintenance conditions [3]; its aberrant reactivation is known to be related to the development of several human malignancies, i.e. basal cell carcinoma (BCC) and medulloblastoma (MB), thus representing a promising therapeutic target for cancer treatment [4][5][6][7][8]. The main players of the HH pathway include the HH ligands Sonic (SHH), Indian (IHH) or Desert Hedgehog (DHH), the Patched transmembrane receptors (PTCH1 and 2), the G protein-coupled-like receptor Smoothened (SMO), and the glioma-associated oncogene (GLI) transcriptional factors (GLI1, GLI2 and GLI3).…”