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<p>Current analytical techniques are limited in their ability to interrogate the mechanistic relationship between aberrant mucin glycosylation and malignancy. Herein, we describe a method for mapping specific mucin glycoforms in diseased tissue, enabling correlation of the tumor glycan profile with malignant features. Following on-tissue digestion with mucinase StcE, matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) deduces the spatial distribution of mucinous O-glycopeptides that are subsequently identified using liquid chromatography coupled to mass spectrometry (LC-MS). Our coupled MS approach reveals a striking mucin 2 (MUC2) expression pattern in three colorectal mucinous adenocarcinomas, in which different glycoforms clearly stratify regions within each tumor. The LC-MS experiments obtain glycoproteomic sequence coverage of MUC2’s mucin domains in unprecedented depth, and we also present evidence for an endogenous O-acetylated GalNAc. Overall, this proof-of-concept work underscores the potential of this technique to drive research in oncology and beyond.</p>

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Glycosite mapping and in situ mass spectrometry imaging of MUC2 glycopeptides via on-slide mucinase digestion

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年5月7日

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  • Sarah C. Lowery,
  • Mikaela K. Ribi,
  • Joann Chongsaritsinsuk,
  • Isabella P. Tran,
  • Grace Grimsley,
  • Rachel Stubler,
  • Keira E. Mahoney,
  • Taryn M. Lucas,
  • Georgia Charkoftaki,
  • Alvaro Santos-Neto,
  • Nissi Varki,
  • Vasilis Vasiliou,
  • Michael Angelo,
  • Richard R. Drake,
  • Stacy A. Malaker

摘要

Current analytical techniques are limited in their ability to interrogate the mechanistic relationship between aberrant mucin glycosylation and malignancy. Herein, we describe a method for mapping specific mucin glycoforms in diseased tissue, enabling correlation of the tumor glycan profile with malignant features. Following on-tissue digestion with mucinase StcE, matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) deduces the spatial distribution of mucinous O-glycopeptides that are subsequently identified using liquid chromatography coupled to mass spectrometry (LC-MS). Our coupled MS approach reveals a striking mucin 2 (MUC2) expression pattern in three colorectal mucinous adenocarcinomas, in which different glycoforms clearly stratify regions within each tumor. The LC-MS experiments obtain glycoproteomic sequence coverage of MUC2’s mucin domains in unprecedented depth, and we also present evidence for an endogenous O-acetylated GalNAc. Overall, this proof-of-concept work underscores the potential of this technique to drive research in oncology and beyond.