Characterization of Glycocalyx for Biomarker Discovery in Cancer by Liquid Chromatography-Mass Spectrometry Methods
- Zhou, Qingwen
- Advisor(s): Lebrilla, Carlito B.
Glycosylation is one of the most prevalent post-translational modifications found on biomolecules that regulate various biological functions such as cell-cell interactions, immune responses, cellular regulations, viral/bacterial infections, and much more. Aberration in glycans can lead to alterations in protein function and have been shown to occur in many diseases such as cancer. In recent years, a few glycan-based biomarkers for cancer have been discovered. However, cancer is still a leading cause of death worldwide. Therefore, additional effort is needed to understand the cause of cancer and how glycosylation plays a role in cancer prognosis. This dissertation presents a series of state-of-the-art nano-liquid chromatography-mass spectrometry (nanoLC-MS) methods and bioinformatics tools to characterize the glycocalyx. In Chapter I, a thorough introduction of glycan synthesis, glycan biological functions, and methods of glycan analysis are presented. Chapter II presents the alteration of the N-glycome of several cancer cell lines through metabolic engineering using glycosylation inhibitors. Chapter III presents the first extensive “omic” analysis of the glycocalyx of 3D cell culture of cancer to elucidate alteration in the glycocalyx relative to its 2D counterpart. Finally, chapter IV presents the characterization of glycocalyx of lung cancer cell lines treated with a therapeutic drug to monitor glycocalyx alteration.