Abstract 2442: Identification of a novel role for the β-catenin/CBP signaling in epigenetic regulation of the N-glycosylation gene, DPAGT1, in head and neck cancer

Abstract

Abstract Aberrant activation of the DPAGT1 gene, encoding an essential enzyme in the metabolic pathway of protein N-glycosylation, has been shown to be associated with head and neck squamous cell carcinoma (HNSCC). We have shown that DPAGT1 inhibits intercellular adhesion and functions in a positive feedback loop with Wnt/β-catenin signaling, and that the nuclear β-catenin/CBP signaling underlies the progression of HNSCC to advanced disease. However, the tumor promoting effects of DPAGT1 and its molecular links to the nuclear β-catenin/CBP axis are not well defined. We carried out genomic and functional analyses of DPAGT1 perturbation in indolent (CAL27) and metastatic (HSC-3) HNSCC cells, and in orthotopic HSC-3-derived xenografts in mice. We further generated and annotated DPAGT1 inhibition signature in HSC-3 cells and interrogated it in TCGA HNSCC. We then examined the effects of inhibition of β-catenin-CBP interaction with E7386 on DPAGT1 expression using ChIP-seq and computational approaches. E7386, a novel β-catenin/CBP modulator displays activity profile that closely overlaps with that of ICG-001, but exhibits ~50 - 100-fold lower EC50 values. Ectopic expression of DPAGT1 in indolent CAL27 cells induced epithelial-to-mesenchymal transition (EMT) which coincided with increased abundance of active β-catenin. Partial knockdown of DPAGT1 with siRNA in metastatic HSC-3 cells inhibited EMT, diminished cell migration and enhanced intercellular adhesion. Inhibition of the DPAGT1 enzyme activity using tunicamycin interfered with orthotopic tongue tumor growth and metastasis. DPAGT1 knockdown in HSC-3 cells defined DPAGT1-activated gene signature as enriched in pro-tumorigenic signaling pathways, including stem cell-like genes. Integrative analysis of the DPAGT1-activated genes in TCGA validated the association of DPAGT1 activity with the EMT transcription factors, ZEB1, Twist1/2 and Slug. ChIP-seq analyses without and with the E7386 treatment revealed reduced occupancy of H3K4me3 at two DPAGT1 transcription start sites following the E7386 treatment. In conclusion, our studies align aberrant activation of DPAGT1 with the induction of EMT and stem cell associated genes and suggest a novel role of β-catenin/CBP/MLL1 in the epigenetic regulation DPAGT1 and protein N-glycosylation in HNSCC. Citation Format: Khalid A. Alamoud, Vinay Kartha, Huamei Yang, Andrew Tilston-Lunel, Anthony Federico, Manish Bais, Takachi Owa, Kenichi Nomoto, Xalarabos Varelas, Stefano Monti, Maria A. Kukuruzinska. Identification of a novel role for the β-catenin/CBP signaling in epigenetic regulation of the N-glycosylation gene, DPAGT1, in head and neck cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2442.