Abstract
We have shown that Raman microspectroscopy is a powerful method for visualization of glycocalyx offering cellular interrogation without staining, unprecedented spatial and spectral resolution, and biochemical information. We showed for the first time that Raman imaging can be used to distinguish successfully between glycosylated and nonglycosylated proteins in normal and cancer tissue. Thousands of protein, lipid and glycan species exist in cells and tissues and their metabolism is monitored via numerous pathways, networks and methods. The metabolism can change in response to cellular environment alterations, such as development of a disease. Measuring such alterations and understanding the pathways involved are crucial to fully understand cellular metabolism in cancer development. In this paper Raman markers of glycogen, glycosaminoglycan, chondroitin sulfate, heparan sulfate proteoglycan were identified based on their vibrational signatures. High spatial resolution of Raman imaging combined with chemometrics allows separation of individual species from many chemical components present in each cell. We have found that metabolism of proteins, lipids and glycans is markedly deregulated in breast (adenocarcinoma) and brain (medulloblastoma) tumors. We have identified two glycoforms in the normal breast tissue and the malignant brain tissue in contrast to the breast cancer tissue where only one glycoform has been identified.
Highlights
The most important components of cells are proteins, lipids and carbohydrates
We will examine if Raman spectroscopy and Raman imaging are useful to define all of the molecular species, glycoforms of glycoproteins, in a cell or tissue
We have shown that Raman microspectroscopy is a powerful method for visualization of glycocalyx offering cellular interrogation without staining, unprecedented spatial and spectral resolution to provide important biochemical information
Summary
The most important components of cells are proteins, lipids and carbohydrates. Lipids and carbohydrates are used primarily as source of energy, but it has been shown recently that they move along intersecting sets of metabolic pathways with impact on human diseases, such as cancer[1,2,3,4,5]. Recent advances in glycomics reveal the scope and scale of their functional roles and their impact on human disease[2,3]. Glycosaminoglycans (GAGs) are important subgroup of glycans, because they are major components of the extracellular matrix in human tissues, and play important roles in various physiological processes, such as lubrication, a shock absorption in the tissue, supporting collagen and elastin in the cellular spaces and keeping protein fibers in balance.
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