B3GALT4 remodels the tumor microenvironment through GD2-mediated lipid raft formation and the c-met/AKT/mTOR/IRF-1 axis in neuroblastoma

Abstract

Background Beta-1,3-galactosyltransferase-4 (B3GALT4) plays a critical regulatory role in tumor biology. However, the role of B3GALT4 in modulating the tumor microenvironment (TME) of neuroblastoma (NB) remains unknown. Methods Public datasets and clinical NB samples were collected to evaluate the expression and clinical significance of GD2 and B3GALT4 in NB patients. CCK-8, colony formation, and transwell assays and experiments in tumor-bearing mouse models were conducted to investigate the function of B3GALT4. Flow cytometry, ELISA, immunohistochemistry, immunofluorescence, western blotting, and chemotaxis assays were conducted to ascertain the immunomodulatory mechanism of B3GALT4. The combined therapeutic effect of the lipid raft inhibitor MβCD and anti-GD2 mAb was validated in a murine model of NB. Results GD2 was overexpressed in NB tissues and high expression of GD2 was associated with poor prognosis in NB patients. B3GALT4 was downregulated in NB tissues, and low expression of B3GALT4 indicated poor prognosis in NB patients. Silencing B3GALT4 significantly enhanced tumor progression both in vitro and in vivo. Meanwhile, the overexpression of B3GALT4 increased the recruitment of CD8+ T lymphocytes via the chemokines CXCL9 and CXCL10. Additionally, B3GALT4 regulated NB-cell GD2 expression and lipid raft formation. Mechanistically, B3GALT4 regulated the expression of CXCL9 and CXCL10 via the c-Met signaling in the lipid rafts and the downstream AKT/mTOR/IRF-1 pathway. The lipid raft inhibitor, MβCD, attenuated B3GALT4 deficiency-induced tumor progression and immune evasion. Last, MβCD combined with anti-GD2 mAb treatment significantly enhanced the antitumor effect and the infiltration of CD8+ T cells. Conclusions Upregulation of B3GALT4 promotes the secretion of CXCL9 and CXCL10 to recruit CD8+ T lymphocytes via the GD2-mediated lipid rafts and the c-Met/AKT/mTOR/IRF-1 pathway. Moreover, lipid raft inhibitors may enhance the efficacy of anti-GD2 immunotherapy for NB.

Optimal combination of MYCN differential gene and cellular senescence gene predicts adverse outcomes in patients with neuroblastoma

“…Studies on the TME have demonstrated that the formation of an immunosuppressive TME contributes to the poor prognosis of NB patients and the limited effectiveness of immunotherapy ( 42 , 43 ). In a previous study conducted by our group, we successfully reconstructed normal blood vessels in a mouse model of NB using a multi-target tyrosine kinase inhibitor (TKI) called Anlotinib ( 9 ).…”

Biology of GD2 ganglioside: implications for cancer immunotherapy

“…As will be seen in further details later in this work, there is a tissue- and age-dependent restriction of GD2 expression ( Svennerholm et al, 1991 ; Lammie et al, 1993 ; Huang et al, 2022 ). GD2 biosynthesis is mostly regulated by the activity of different glycosyltransferases, with emphasis on ST8SIA1, B4GALNT1 & B3GALT4 ( Sorokin et al, 2020 ; Sha et al, 2022 ), and is controlled epigenetically, transcriptionally, and post-transcriptionally.…”

“…This suggests additional levels of regulation differentiating normal from aberrant cells. This distinction can be a result of varying transcriptional regulations in the three genes mainly responsible for GD2 expression ( Sorokin et al, 2020 ; Sha et al, 2022 ). This additionally means that GD2 can be a useful stage-specific marker molecule of developing cells.…”

“…GD2 overexpression can be further correlated with the upregulation of ST8SIA1 and B4GALNT1, or the downregulation of B3GALT4 ( Sorokin et al, 2020 ; Yoshida et al, 2020 ; Kasprowicz et al, 2022 ; Mabe et al, 2022 ; Sha et al, 2022 ). So far, no mutation has been reported for these genes, and the specific causes of GD2 overexpression remain unclear.…”

“…At the plasma membrane level, GD2 has been identified as a lipid raft component ( Sha et al, 2022 ). Lipid rafts are specialized, sphingolipid-enriched, dynamic microdomains within the membrane that can compartmentalize cellular processes.…”