Peripheral neuroblastic tumors (pNTs) are the most common childhood extracranial solid tumors. There are several therapeutic strategies targeting disialoganglioside GD2. Disialoganglioside GD3 has become a potential target. However, the mechanism by which pNTs express GD3 and GD2 remains unclear. We investigated the combined expression status of GD3 and GD2 in pNTs and delineated their clinicopathological values.
Methods
GD3 and GD2 expression was examined in pNT tissue samples (n = 35) using immunohistochemistry and multiple immunofluorescence imaging.
Results
GD3 and GD2 expression was positive in 32/35 and 25/35 samples, respectively. Combinatorial analysis of GD3 and GD2 expression in neuroblastoma showed that both were heterogeneously expressed from cell to cell. There were higher numbers of GD3-positive and GD2-negative cells in the low-risk group than in the intermediate-risk (P = 0.014) and high-risk (P = 0.009) groups. Cases with high proportions of GD3-positive and GD2-negative cells were associated with the International Neuroblastoma Staging System stage (P = 0.004), Children’s Oncology Group risk group (P = 0.001), and outcome (P = 0.019) and tended to have a higher overall survival rate.
Conclusion
We demonstrated that neuroblastomas from low-risk patients included more GD3-positive and GD2-negative cells than those from high-risk patients. Clarifying the heterogeneity of neuroblastoma aids in better understanding the biological characteristics and clinical behavior.
ShimadaHAmbrosIMDehnerLPHataJJoshiVV.The International Neuroblastoma Pathology Classification (the Shimada system). Cancer. 1999;86(2):364–372.
4.
SchmidtM LouLalASeegerRC, et al. Favorable prognosis for patients 12 to 18 months of age with stage 4 nonamplified MYCN neuroblastoma: a Children’s Cancer Group Study.J Clin Oncol. 2005;23:6474–6480.
5.
LammieGACheungNKVGeraldWRosenblumMCordon-CardoC.Ganglioside GD2 expression in the human nervous system and in neuroblastomas – An immunohistochemical study. Int J Oncol. 1993;3:909–915.
6.
NazhaBInalCOwonikokoTK.Disialoganglioside GD2 expression in solid tumors and role as a target for cancer therapy. Front Oncol. 2020;10:1–15.
7.
MatthayKKVillablancaJGSeegerRC, et al. Treatment of high-risk neuroblastoma with intensive chemotherapy, radiotherapy, autologous bone marrow transplantation, and 13-cis-retinoic acid. N Engl J Med. 1999;341:1165–1173.
8.
PortoukalianJDavidM‐JRichardM, et al. Shedding of GD2 ganglioside in patients with retinoblastoma.Int J Cancer. 1993;53:948–951.
9.
MountCWMajznerRGSundareshS, et al. Potent antitumor efficacy of anti-GD2 CAR T-cells in H3K27M+ diffuse midline gliomas Christopher. Nat Med. 2018;24:572–579.
10.
DobrenkovKOstrovnayaIGuJ, et al. Oncotargets GD2 and GD3 are highly expressed in sarcomas of children, adolescents, and young adults. Pediatr Blood Cancer. 2016;63(10):1780–1785.
11.
JulienSBobowskiMSteenackersALe BourhisXDelannoyP.How do gangliosides regulate RTKs signaling?Cells. 2013;2:751–767.
12.
MatthayKKReynoldsCPSeegerRC, et al. Long-term results for children with high-risk neuroblastoma treated on a randomized trial of myeloablative therapy followed by 13-cis-retinoic acid: a Children’s Oncology Group Study. J Clin Oncol. 2009;27:1007–1013.
13.
CohnSLPearsonADJLondonWB, et al. The International Neuroblastoma Risk Group (INRG) classification system: an INRG task force report. J Clin Oncol. 2009;27:289–297.
14.
MatthayKKGeorgeREYuAL.Promising therapeutic targets in neuroblastoma. Clin Cancer Res. 2012;18:2740–2753.
15.
TerzicTCordeauMHerblotS, et al. Expression of disialoganglioside (GD2) in neuroblastic tumors: a prognostic value for patients treated with anti-GD2 immunotherapy.Pediatr Dev Pathol. 2018;21:355–362.
16.
OhmiYKambeMOhkawaY, et al. Differential roles of gangliosides in malignant properties of melanomas. PLoS One. 2018;13:1–23.
17.
IwasawaTZhangPOhkawaY, et al. Enhancement of malignant properties of human glioma cells by ganglioside GD3/GD2. Int J Oncol. 2018;52:1255–1266.
18.
WebbTJLiXGiuntoliRL, et al. Molecular identification of GD3 as a suppressor of the innate immune response in ovarian cancer.Cancer Res. 2013;72:3744–3752.
19.
LeeMKimKSFukushiAKimDHKimCHLeeYC.Transcriptional activation of human GD3 synthase (hST8sia i) gene in curcumin-induced autophagy in A549 human lung carcinoma cells. Int J Mol Sci. 2018;19:1–11.
20.
WagenerRRöhnGSchillingerGSchröderRKobbeBErnestusRI.Ganglioside profiles in human gliomas: quantification by microbore high performance liquid chromatography and correlation to histomorphology and grading. Acta Neurochir (Wien). 1999;141:1339–1345.
21.
AllredDCHarveyJMClarkGM.Prognostic and predictive factors in breast cancer by immunohistochemical analysis. Mod Pathol. 1998;11:155–168.
22.
NakanishiYShimizuTTsujinoI, et al. Semi-nested real-time reverse transcription polymerase chain reaction methods for the successful quantitation of cytokeratin mRNA expression levels for the subtyping of non-small-cell lung carcinoma using paraffin-embedded and microdissected lung biopsys. Acta Histochem Cytochem. 2013;46:85–96.
23.
Macabeo-OngMGinzingerDGDekkerN, et al. Effect of duration of fixation on quantitative reverse transcription polymerase chain reaction analyses.Mod Pathol. 2002;15:979–987.
24.
CaseyDLCheungNKV.Immunotherapy of pediatric solid tumors: treatments at a crossroads, with an emphasis on antibodies.Cancer Immunol Res. 2020;8:161–166.
25.
HamamuraKTsujiMHottaH, et al. Functional activation of Src family kinase yes protein is essential for the enhanced malignant properties of human melanoma cells expressing ganglioside GD3.J Biol Chem. 2011;286:18526–18537.
26.
HedbergKMDellhedenBWikstrandCJFredmanP.Monoclonal anti-GD3 antibodies selectively inhibit the proliferation of human malignant glioma cells in vitro. Glycoconj J. 2000;10:717–726.
27.
KangNYKimCHKimKS, et al. Expression of the human CMP-NeuAc:GM3 α2,8-sialyltransferase (GD3 synthase) gene through the NF-κB activation in human melanoma SK-MEL-2 cells.Biochim Biophys Acta – Gene Struct Expr. 2007;1769:622–630.
28.
SarkarTRBattulaVLWerdenSJ, et al. GD3 synthase regulates epithelial-mesenchymal transition and metastasis in breast cancer. Oncogene. 2015;34:2958–2967.
29.
LluisJMLlacunaLvon MontfortC, et al. GD3 synthase overexpression sensitizes hepatocarcinoma cells to hypoxia and reduces tumor growth by suppressing the cSrc/NF-κb survival pathway. PLoS One. 2009;4:e8059.
OblingerJLPearlDKBoardmanCL, et al. Diagnostic and prognostic value of glycosyltransferase mRNA in glioblastoma multiforme patients. Neuropathol Appl Neurobiol. 2006;32:410–418.
34.
DippoldWGKnuthAZum BuschenfeldeKHMDienesHP.Immunohistochemical localization of ganglioside GD3 in human malignant melanoma, epithelial tumors, and normal tissues. Cancer Res. 1985;45:3699–3705.
35.
KushnerBHCheungIYModakSKramerKRagupathiGCheungNKV.Phase I trial of a bivalent gangliosides vaccine in combination with β-glucan for high-risk neuroblastoma in second or later remission. Clin Cancer Res. 2014;20:1375–1382.
36.
RossigCKailayangiriSJamitzkySAltvaterB.Carbohydrate targets for CAR T cells in solid childhood cancers. Front Oncol. 2018;8:1–12.
37.
DoroninIIVishnyakovaPAKholodenkoIV, et al. Ganglioside GD2 in reception and transduction of cell death signal in tumor cells. BMC Cancer. 2014;14:295.
38.
ShibuyaHHamamuraKHottaH, et al. Enhancement of malignant properties of human osteosarcoma cells with disialyl gangliosides GD2/GD3. Cancer Sci. 2012;103:1656–1664.
39.
OhkawaYMiyazakiSHamamuraK, et al. Ganglioside GD3 enhances adhesion signals and augments malignant properties of melanoma cells by recruiting integrins to glycolipid-enriched microdomains. J Biol Chem. 2010;285:27213–27223.
40.
SwertsKAmbrosPFBrouzesC, et al. Standardization of the immunocytochemical detection of neuroblastoma cells in bone marrow. J Histochem Cytochem. 2005;53:1433–1440.
41.
BeiskeKBurchillSACheungIY, et al, Consensus criteria for sensitive detection of minimal neuroblastoma cells in bone marrow, blood and stem cell preparations by immunocytology and QRT-PCR: recommendations by the International Neuroblastoma Risk Group Task Force. Br J Cancer. 2009;19:1627–1637.
