C1GALT1 expression predicts a favorable prognosis and suppresses malignant phenotypes via TrkA signaling in neuroblastoma

Neng-Yu Lin,Min-Chuan Huang,Shiann-Tarng Jou,Shu-Wei Chou,Kai-Hsin Lin,Wen-Ming Hsu,Dong-Tsamn Lin,Yung-Li Yang,Meng-Yao Lu,Hsiu-Ling Chang,Hsiu-Hao Chang,Syue-Ting Chen

doi:10.1038/s41389-022-00383-w

Abstract

Neuroblastoma (NB) is a childhood tumor derived from the sympathoadrenal lineage of the neural crest progenitor cells. Core 1 β1,3-galactosyltransferase (C1GALT1) controls the crucial step of GalNAc-type O-glycosylation, and its altered expression affects cancer behaviors. However, the role of C1GALT1 in NB tumors remains unclear. Our data showed that C1GALT1 expression was significantly associated with differentiated tumor histology, correlated with TrkA expression, and predicted good prognosis independently in NB. Downregulation of C1GALT1 promotes malignant behaviors of NB cells in vitro and in vivo. Mechanistic investigation showed that knockdown of C1GALT1 in NB cells increased TrkA pulled down through Vicia villosa agglutinin beads, indicating the modulation of O-glycans on TrkA by C1GALT1, and silencing C1GALT1 suppressed the TrkA expression on the NB cell surface. Overexpression of C1GALT1 increased the protein levels of TrkA and promoted the differentiation of NB cells, whereas knockdown of TrkA inhibited C1GALT1-induced neuronal differentiation. Moreover, the inhibitory effects of migration and invasion in C1GALT1-overexpressing NB cells were blocked by TrkA downregulation. C1GALT1 knockdown enhanced AKT phosphorylation but attenuated ERK phosphorylation, and these properties were consistent in C1GALT1-overexpressing NB cells with TrkA knockdown. Taken together, our data provided the first evidence for the existence of GalNAc-type O-glycans on TrkA and altered O-glycan structures by C1GALT1 can regulate TrkA signaling in NB cells. This study sheds light on the novel prognostic role of C1GALT1 in NB and provides new information of C1GALT1 and TrkA on the pathogenesis of NB.

Highlights

Neuroblastoma (NB) is a childhood tumor derived from sympathoadrenal lineage of the neural crest progenitor cells
C1GALT1-mediated O-glycans regulate cell surface expression and signaling of TrkA in NB cells We have identified that multiple receptor tyrosine kinase (RTKs), such as EGFR, FGFR2, EPHA2, and MET, are key targets for C1GALT1 and their appropriate O-glycosylation is essential for cancer progression and metastasis [24, 26–28]
The results showed that knockdown of C1GALT1 in NB cells increased TrkA pulled down through Vicia villosa agglutinin (VVA) and Griffonia simplicifolia lectin I (GSL-I) lectin beads

Summary

INTRODUCTION

Neuroblastoma (NB) is a childhood tumor derived from sympathoadrenal lineage of the neural crest progenitor cells. Yung-Feng Liao, Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan These cell lines were cultured with Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen) containing 10% FBS (Invitrogen), 100 IU/mL penicillin, and 100 μg/mL streptomycin (Invitrogen) in a humidified tissue culture incubator at 37 °C and 5% CO2 atmosphere. The present study is the first report to show that high C1GALT1 expression correlates with NB tumor differentiation status and predicts better survival outcomes of patients with NB. MYCN status of the tumor tissue was evaluated by chromogenic in situ hybridization analysis of formalin-fixed paraffinembedded tissues or fresh tumor single cells [33] The treatment of these patients has been described in previous studies [34–36]. Immunofluorescence microscopy Cells were cultured in chamber slides (SPL Life Sciences), fixed with 4% PFA, permeabilized with 0.25% Triton X100, blocked in PBS containing 2% bovine serum albumin (Bio-Rad) and applied with primary antibodies to TrkA (LSBio), Lamp (GeneTex).

RESULTS

DISCUSSION

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