The Prevalence and Nature of Glycan Alterations on Specific Proteins in Pancreatic Cancer Patients Revealed Using Antibody-Lectin Sandwich Arrays

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Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. Glycomic and glycoproteomic profiling studies have reported extensive protein glycosylation pattern alterations in ovarian cancer. Therefore, spatio-temporal investigation of these glycosylation changes may unearth tissue-specific changes that occur in the development and progression of ovarian cancer. A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffinembedded ( Ovarian cancer is the fifth most fatal malignancy in adult women with an estimated 21,290 new cases diagnosed and 14,180 deaths recorded in the United States during 2015 (1). There are several reasons for the poor prognosis of ovarian cancer and its diagnosis at advanced stage-lack of diagnostic markers for the early detection (2, 3), rapid metastasis of the disease (4), and limited or modest understanding of the etiology, origin and the diverse clinical and pathological behavior of the tumors (5). Moreover, epithelial ovarian cancer comprises of several distinct sub-types based on their histopathological features into serous, endometrioid, clear-cell, mucinous, and undifferentiated subtypes (6, 7).Protein glycosylation is an important post-translational modification which has relevance in many biological processes such as cell signaling, immune responses, extracellular interaction and cell adhesion (8, 9). Aberrant protein glycosylation such as the expression of truncated glycans as well From the ‡Faculty

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confidence: 99%

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confidence: 99%

N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues

Everest‐Dass

Briggs

Kaur

et al. 2016

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132

“…In the further pursuit of alternative molecular markers, the patterns of glycosylation have been studied in sera of patients with pancreatic cancer using antibody-and lectin-based sensors (7,8). A similar approach has been used to study pancreatic cyst fluids, targeting representative glycans on prominent candidate glycoprotein markers, including several of the mucins and carcinoembryonic antigen-related cell adhesion molecules.…”

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confidence: 99%

Glycomic and Proteomic Profiling of Pancreatic Cyst Fluids Identifies Hyperfucosylated Lactosamines on the N-linked Glycans of Overexpressed Glycoproteins

Mann

Goetz

House

et al. 2012

Pancreatic cancer is now the fourth leading cause of cancer deaths in the United States, and it is associated with an alarmingly low 5-year survival rate of 5%. However, a patient's prognosis is considerably improved when the malignant lesions are identified at an early stage of the disease and removed by surgical resection. Unfortunately, the absence of a practical screening strategy and clinical diagnostic test for identifying premalignant lesions within the pancreas often prevents early detection of pancreatic cancer. To aid in the development of a molecular screening system for early detection of the disease, we have performed glycomic and glycoproteomic profiling experiments on 21 pancreatic cyst fluid samples, including fluids from mucinous cystic neoplasms and intraductal papillary mucinous neoplasms, two types of mucinous cysts that are considered high risk to undergo malignant transformation. A total of 80 asparagine-linked (N-linked) glycans, including high mannose and complex structures, were identified. Of special interest was a series of complex N-linked glycans containing two to six fucose residues, located predominantly as substituents on ␤-lactosamine extensions. Following the observation of these "hyperfucosylated" glycans, bottom-up proteomics experiments utilizing a label-free quantitative approach were applied to the investigation of two sets of tryptically digested proteins derived from the cyst fluids: 1) all soluble proteins in the raw samples and 2) a subproteome of the soluble cyst fluid proteins that were selectively enriched for fucosylation through the use of surface-immobilized Aleuria aurantia lectin. A comparative analysis of these two proteomic data sets identified glycoproteins that were significantly enriched by lectin affinity. Several candidate glycoproteins that appear hyperfucosylated were identified, including triacylglycerol lipase and pancreatic ␣-amylase, which were 20-and 22-fold more abundant, respectively, following A. aurantia lectin enrichment. Molecular & Cellular Proteomics 11: 10.1074/mcp.M111.015792, 1-11, 2012.Pancreatic cancer is associated with a poor long term outcome with an actual 5-year survival rate of less than 5%. The absence of a practical screening strategy and clinical diagnostic test for identifying premalignant lesions within the pancreas prevents early detection of pancreatic cancer. Detection of early stage pancreatic cancer will likely result from novel clinical methods aimed at early tumor detection and molecular profiling of individuals at high risk for pancreatic carcinogenesis.High-resolution imaging techniques have increased the detection of small, organ-based lesions in the intra-abdominal cavity. Incidental cysts within the pancreas are detected in ϳ13% of patients undergoing cross-sectional imaging for nonspecific abdominal symptoms (1). Greater than 80% of these incidental cysts are non-neoplastic, so few carry malignant potential. Pancreatic cysts are divided into three broad pathologic entities: serous cystadenomas (SCA), 1 mucinous ...

“…A glycoform of MUC5AC, one that binds the lectin wheat germ agglutinin (WGA), was almost exclusively found in the mucinous cysts and was a better biomarker than the MUC5AC protein measured over all glycoforms. Furthermore, the glycosylation patterns in pancreatic cysts were different from those we found in pancreatic ductal adenocarcinomas (21,22).…”

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confidence: 37%

Specific Glycoforms of MUC5AC and Endorepellin Accurately Distinguish Mucinous from Nonmucinous Pancreatic Cysts

Cao

Maupin

Curnutte

et al. 2013

Self Cite

Specific protein glycoforms may be uniquely informative about the pathological state of a cyst and may serve as accurate biomarkers. Here we tested that hypothesis using antibody-lectin sandwich arrays in broad screens of protein glycoforms and in targeted studies of candidate markers. We profiled 16 different glycoforms of proteins captured by 72 different antibodies in cyst fluid from mucinous and nonmucinous cysts (n ‫؍‬ 22), and we then tested a three-marker panel in 22 addition samples and 22 blinded samples. Glycan alterations were not widespread among the proteins and were mainly confined to MUC5AC and endorepellin. Specific glycoforms of these proteins, defined by reactivity with wheat germ agglutinin and a blood group H antibody, were significantly elevated in mucinous cysts, whereas the core protein levels were not significantly elevated. A three-marker panel based on these glycoforms distinguished mucinous from nonmucinous cysts with 93% accuracy (89% sensitivity, 100% specificity) in a prevalidation sample set (n ‫؍‬ 44) and with 91% accuracy (87% sensitivity, 100% specificity) in independent, blinded samples (n ‫؍‬ 22). Targeted lectin measurements and mass spectrometry analyses indicated that the higher wheat germ agglutinin and blood group H reactivity was due to oligosaccharides terminating in GlcNAc or N-acetyl-lactosamine with occasional ␣1,2-linked fucose. The results show that MUC5AC and endorepellin glycoforms may be highly specific and sensitive biomarkers for the differentiation of mucinous from nonmucinous pancreatic cysts. Molecular & Cellular