O2161: Fucosyltransferase Blockade is a Novel Metabolic Vulnerability in MYCN-amplified NBs

Background: High-risk neuroblastomas (NBs) have less than 50% overall survival despite aggressive multimodal treatment strategies, highlighting the need to develop new therapeutics. Glycosylation is an underappreciated mediator of cancer progression. We have previously reported enrichment of core fucosylated glycans in MYCN-amplified human NB tissues using matrix-assisted laser desorption ionization mass spectroscopy imaging (MALDI-MSI). 2-FF is a small molecule inhibitor of fucosyltransferases. Herein, we hypothesized that small molecule fucosyltransferase blockade would impede MYCN-amplified NB progression.

Methods: Kaplan-meier analysis was performed to determine whether specific fucosyltransferase expression was associated with overall survival. Core fucosylated glycan abundance was measured by western blotting, ELISA, and flow cytometry using Aleuria aurantia lectin (AAL). Cellular adhesion was measured in vitro using a Vybrant blue adhesion assay. Cyclin D1 and PARP cleavage were quantified by western blotting for markers of proliferation and apoptosis induction. Subcutaneous tumor formation using MYCN-amp BE(2)-C cells was our measure of in vivo tumorigenesis (UK IACUC #2021-3934). For small molecule trials, mice with established BE(2)-C tumors were randomized to receive water-supplemented with 2-FF or vehicle control.

Results: High expression of fucosyltransferase 5 (FUT5; bonf P= 2.57x10-4) and fucosyltransferase 8 (FUT8; bonf P= 5.95x10-3) are associated with poor overall patient survival. 2-FF blocks NB core fucosylation, and NB cell growth and adherence in vitro. 2-FF induces also induces PARP cleavage and cell cycle arrest detected by western blotting. Oral 2-FF administration blocks core fucosylation in vivo and impedes established tumor progression ( < 0.05; Fig. 1). Histologic analysis revealed enhanced induction of cell necrosis (51%vs.12%; p< 0.0001) within treated tumor samples (Fig.2).

Conclusion: Elevated fucosyltransferase 5 and 8 expression are associated with poor overall survival in human NB samples. Small molecule inhibition with 2-FF blocks NB cell growth and adherence in vitro and abrogates tumor growth via the induction of cancer cell death in vivo. These critical findings identify fucosyltransferase blockade as a novel metabolic vulnerability that may be exploited in designing novel treatment paradigms for high-risk NBs.