Global analysis of human glycosyltransferases reveals novel targets for pancreatic cancer pathogenesis

Rohitesh Gupta, Frank Leon, Christopher M. Thompson, Ramakrishna Nimmakayala, Saswati Karmakar, Palanisamy Nallasamy, Seema Chugh, Dipakkumar R. Prajapati, Satyanarayana Rachagani, Sushil Kumar, Moorthy P. Ponnusamy

Research output: Contribution to journalArticle

Abstract

Background: Several reports have shown the role of glycosylation in pancreatic cancer (PC), but a global systematic screening of specific glycosyltransferases (glycoTs) in its progression remains unknown. Methods: We demonstrate a rigorous top-down approach using TCGA-based RNA-Seq analysis, multi-step validation using RT-qPCR, immunoblots and immunohistochemistry. We identified six unique glycoTs (B3GNT3, B4GALNT3, FUT3, FUT6, GCNT3 and MGAT3) in PC pathogenesis and studied their function using CRISPR/Cas9-based KD systems. Results: Serial metastatic in vitro models using T3M4 and HPAF/CD18, generated in house, exhibited decreases in B3GNT3, FUT3 and GCNT3 expression on increasing metastatic potential. Immunohistochemistry identified clinical significance for GCNT3, B4GALNT3 and MGAT3 in PC. Furthermore, the effects of B3GNT3, FUT3, GCNT3 and MGAT3 were shown on proliferation, migration, EMT and stem cell markers in CD18 cell line. Talniflumate, GCNT3 inhibitor, reduced colony formation and migration in T3M4 and CD18 cells. Moreover, we found that loss of GCNT3 suppresses PC progression and metastasis by downregulating cell cycle genes and β-catenin/MUC4 axis. For GCNT3, proteomics revealed downregulation of MUC5AC, MUC1, MUC5B including many other proteins. Conclusions: Collectively, we demonstrate a critical role of O- and N-linked glycoTs in PC progression and delineate the mechanism encompassing the role of GCNT3 in PC.

Original languageEnglish (US)
Pages (from-to)1661-1672
Number of pages12
JournalBritish journal of cancer
Volume122
Issue number11
DOIs
StatePublished - May 26 2020

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Fingerprint Dive into the research topics of 'Global analysis of human glycosyltransferases reveals novel targets for pancreatic cancer pathogenesis'. Together they form a unique fingerprint.

  • Cite this

    Gupta, R., Leon, F., Thompson, C. M., Nimmakayala, R., Karmakar, S., Nallasamy, P., Chugh, S., Prajapati, D. R., Rachagani, S., Kumar, S., & Ponnusamy, M. P. (2020). Global analysis of human glycosyltransferases reveals novel targets for pancreatic cancer pathogenesis. British journal of cancer, 122(11), 1661-1672. https://doi.org/10.1038/s41416-020-0772-3