Fig 1: TERT expression and the ALT pathway are associated with MRS-detectable metabolic reprogramming in LGOG and LGA patient biopsies.TERT mRNA expression (a), telomerase activity (b), ATRX mRNA expression (c), and c-circle level (d) in LGOG (red circles), LGA (magenta circles), and gliosis (blue circles) biopsies. Steady-state metabolite levels measured by 1H-MRS (e) and NADH levels measured by spectrophotometry (f) in ex vivo extracts from LGOG (red circles), LGA (magenta circles), and gliosis (blue circles) biopsies. 2-HG: 2-hydroxyglutarate; PC: phosphocholine; tCreatine: total creatine; NAA: N-acetylaspartate. GLS activity (g), NAMPT activity (h), and expression of ASCT2 and LAT2 (i) in LGOG (red circles), LGA (magenta circles), and gliosis (blue circles) biopsies. All experiments were performed on eight biological replicates each (n = 8). Results are presented as mean ± standard deviation. Statistical significance was assessed using an unpaired Student’s t test assuming unequal variance with p < 0.05 considered significant. Correction for multiple comparisons was performed using the Holm–Šídák method. *** represents statistical significance with p < 0.005. For the sake of clarity, statistical significance with p < 0.005 is represented with # for panel (e). Source data with exact p values are provided as a source data file.
Fig 2: Silencing TERT expression or the ALT pathway normalizes 1H-MRS biomarkers and hyperpolarized [1-13C]-alanine metabolism in patient-derived LGOG and LGA models.a Effect of TERT silencing on steady-state metabolite levels as measured by 1H-MRS in the BT54 LGOG model (BT54 TERT + neurospheres: red circles; BT54 TERT- #1 neurospheres: dark brown circles; BT54 TERT- #2 neurospheres: light brown circles). TERT silencing was carried out with two independent, nonoverlapping siRNA sequences/pools. b Representative summed 13C-MRS spectra showing hyperpolarized [1-13C]-alanine metabolism in BT54 TERT+ and BT54 TERT- neurospheres. c Quantification of the ratios for hyperpolarized lactate/alanine and hyperpolarized pyruvate/alanine in BT54 TERT+ and TERT- neurospheres (BT54 TERT+ neurospheres: red circles; BT54 TERT- #1 neurospheres: dark brown circles; BT54 TERT- #2 neurospheres: light brown circles). TERT silencing was carried out with two independent, nonoverlapping siRNA sequences/pools. d Effect of silencing the ALT pathway via ATRX re-expression on steady-state metabolite levels as measured by 1H-MRS in the BT142 LGA model (BT142 ALT+ neurospheres: magenta circles; BT142 ALT- neurospheres (lavender circles). e Representative summed 13C-MRS spectra showing hyperpolarized [1-13C]-alanine metabolism in BT142 ALT+ and BT142 ALT- neurospheres. f Quantification of the ratios for hyperpolarized pyruvate/alanine and hyperpolarized lactate/alanine in BT142 ALT+ (magenta circles) and BT142 ALT- (lavender circles) neurospheres. All experiments were performed on three biological replicates (n = 3). Results are presented as mean ± standard deviation. Statistical significance was assessed using an unpaired Student’s t test assuming unequal variance with p < 0.05 considered significant. Correction for multiple comparisons was performed using the Holm–Šídák method. *** represents statistical significance with p < 0.005; ** represents statistical significance with p < 0.01; * represents statistical significance with p < 0.05. Also, refer to Supplementary Fig. 4. Source data with exact p values are provided as a source data file.
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