GD2 Expression in Medulloblastoma and Neuroblastoma for Personalized Immunotherapy: A Matter of Subtype

Abstract

Neuroblastoma (NBL) and medulloblastoma (MB) are aggressive pediatric cancers which can benefit from therapies targeting gangliosides. Therefore, we compared the ganglioside profile of 9 MB and 14 NBL samples by thin layer chromatography and mass spectrometry. NBL had the highest expression of GD2 (median 0.54 nmol GD2/mg protein), and also expressed complex gangliosides. GD2-low samples expressed GD1a and were more differentiated. MB mainly expressed GD2 (median 0.032 nmol GD2/mg protein) or GM3. Four sonic hedgehog-activated (SHH) as well as one group 4 and one group 3 MBs were GD2-positive. Two group 3 MB samples were GD2-negative but GM3-positive. N-glycolyl neuraminic acid-containing GM3 was neither detected in NBL nor MB by mass spectrometry. Furthermore, a GD2-phenotype predicting two-gene signature (ST8SIA1 and B4GALNT1) was applied to RNA-Seq datasets, including 86 MBs and validated by qRT-PCR. The signature values were decreased in group 3 and wingless-activated (WNT) compared to SHH and group 4 MBs. These results suggest that while NBL is GD2-positive, only some MB patients can benefit from a GD2-directed therapy. The expression of genes involved in the ganglioside synthesis may allow the identification of GD2-positive MBs. Finally, the ganglioside profile may reflect the differentiation status in NBL and could help to define MB subtypes.

Glucosylceramide Synthase Inhibitors Induce Ceramide Accumulation and Sensitize H3K27 Mutant Diffuse Midline Glioma to Irradiation

“…By using GD2 as a surrogate biomarker of GSI deregulation, we observed that the loss of trimethylation at H3.3K27 in ependymoma was associated with a very high GD2 expression. The expression was even higher than the one observed in a H3K27M-mut DMG and was in the range of the expression described in neuroblastoma [23]. Loss of methylation at H3K27 in ependymoma was observed in the posteriora fossa tumors group A (PFA), which has a poor clinical outcome [66].…”

“…Therefore, tumor tissue from one adult H3K27M-mut DMG patient was used. GD2 amount in the H3K27M-mut DMG sample was not as high as in neuroblastoma [ 23 ]. GD2 expression was rather low in ependymoma; however, the only sample with loss of H3K27me3 showed a very high concentration of GD2, which was even higher than the concentration in the H327M-mut DMG tumor sample.…”

“…Liquid-chromatography-coupled tandem mass spectrometric analysis of sphingolipids. Liquid-chromatography-coupled tandem mass spectrometric analysis of sphingolipids was performed on an Aqcuity I class UPLC (Waters, Milford, MA, USA) coupled through electrospray ionization (ESI) to a triple-quadrupole-like tandem mass spectrometer (Xevo TQ-S, Waters, Milford, MA, USA) according to previous publications [ 23 , 78 ]. As described, GD2 was analyzed with an Anionoic Polar Pesticide column (5 µm, 150 mm × 2.1 mm, Waters) in MRM mode [ 23 ], whereas all other sphingolipids were separated on a reversed-phase CSH C18 column (2.1 mm × 100 mm; 1.7 μm, Waters) in MRM mode [ 78 , 79 ].…”

“…Liquid-chromatography-coupled tandem mass spectrometric analysis of sphingolipids was performed on an Aqcuity I class UPLC (Waters, Milford, MA, USA) coupled through electrospray ionization (ESI) to a triple-quadrupole-like tandem mass spectrometer (Xevo TQ-S, Waters, Milford, MA, USA) according to previous publications [ 23 , 78 ]. As described, GD2 was analyzed with an Anionoic Polar Pesticide column (5 µm, 150 mm × 2.1 mm, Waters) in MRM mode [ 23 ], whereas all other sphingolipids were separated on a reversed-phase CSH C18 column (2.1 mm × 100 mm; 1.7 μm, Waters) in MRM mode [ 78 , 79 ]. In positive electrospray ionization mode protonated molecular ions of S1P, Cer, Cer1P, HexCer (likely GlcCer), and Hex2Cer (likely LacCer), GM3 and GD3 ions were detected with transitions to the product ion m/z 264 and protonated molecular HexNAc-Hex2Cer (likely GA2), GM2, GD2, Tetraosylceramide (likely GA1 and Gb4Cer), and GM1 ions to the product ion m/z 204.…”

“…Additionally, they are associated with key processes of malignant transformation such as tumor growth, differentiation, and the ability to metastasize (reviewed in [ 20 ]). In particular, the disialoganglioside GD2 is characterized by a weak expression in normal tissue, which is predominantly localized to the CNS and peripheral nervous system (PNS) [ 21 ], and a comparatively high expression in solid tumors such as neuroblastoma [ 22 , 23 ] or Ewing’s sarcoma [ 24 , 25 ]. H3K27M-mut DMGs also exhibit strong and homogeneous expressions of GD2 [ 26 ], predisposing them to potential anti-GD2 therapy.…”