Oral Presentation 28th Annual Lorne Proteomics Symposium 2023

Painting a Picture of the Gynaecological Cancer N-Glycome and Proteome (#32)

Matthew T Briggs 1 , Parul Mittal 1 , Manuela Klingler-Hoffmann 1 , Gurjeet Kaur 2 , Martin K Oehler 3 , Peter Hoffmann 1
  1. University of South Australia, Adelaide, SA, Australia
  2. Institute for Research in Molecular Medicine, Universiti Sains , Pulau Pinang, Malaysia
  3. Department of Gynaecological Oncology, Royal Adelaide Hospital, Adelaide, SA, Australia

Gynaecological cancers are a group of tumours that originate from the vulva, vagina, cervix, uterus (endometrium), placenta, fallopian tubes and ovaries in adult women. These cancers are known to undergo epithelial-mesenchymal transition (EMT) which results in the metastasis of epithelial cells and an increase in their resistance to apoptosis by altering the extracellular matrix (ECM). ECM proteins play an imperative role in cell health as they provide the scaffold upon which cells and tissues are built. Hence ECM proteins directly and indirectly influence almost all cellular processes, including cell differentiation, proliferation, and motility. The majority of ECM proteins are known to be heavily glycosylated, most commonly N-glycosylated whereby glycans (i.e., complex sugars) are attached to asparagine residues. Previously, our group has employed MALDI mass spectrometry imaging (MSI) using axial TOF instruments to spatially map N-glycans across formalin-fixed paraffin-embedded (FFPE) ovarian and endometrial cancer tissue sections (1,2). In this study, we have further investigated the N-glycome and proteome of FFPE ovarian, endometrial, cervical and vulvar cancer tissue sections by MALDI-MSI using Bruker’s latest timsTOF fleX instrument. Furthermore, our group has established an in-situ tandem mass spectrometry (MS/MS) fragmentation protocol which can be implemented post-MALDI-MSI analysis to structurally characterise and confirm cancer-specific N-glycans and tryptic peptides. Lastly, our group has successfully separated structural isomers (i.e., N-glycoforms) in-situ using trapped ion mobility mass spectrometry (TIMS). With these complimentary methods in mind, a wealth of information can be acquired from a single tissue section, thereby reducing the sample preparation time previously required while increasing the number of N-glycans and tryptic peptides identified relative to conventional axial TOF instruments. It is with this information that we can better understand glycan and protein alterations within gynaecological cancers in the pursuit of discovering diagnostic or therapeutic targets.

  1. Matthew T. Briggs, Mark R. Condina, Yin Ying Ho, Arun V. Everest-Dass, Parul Mittal, Gurjeet Kaur, Martin K. Oehler, Nicolle H. Packer, Peter Hoffmann (2019). MALDI Mass Spectrometry Imaging of Early- and Late-Stage Serous Ovarian Cancer Tissue Reveals Stage-Specific N-Glycans. Proteomics, 19:21-22, doi.org/10.1002/pmic.201800482.
  2. Parul Mittal, Matthew Briggs, Manuela Klingler-Hoffmann, Gurjeet Kaur, Nicolle H Packer, Martin K Oehler, Peter Hoffmann (2021). Altered N-linked glycosylation in endometrial cancer. Analytical and Bioanalytical Chemistry, 413, 2721-2733, doi.org/10.1007/s00216-020-03039-z