Pancreatic cancer is one of the most deadly forms of human malignancy for which early detection would greatly facilitate increased patient survival. A glyco-microarray approach will be used to search for early detection markers of pancreatic cancer in human plasma. We will use multi-dimensional liquid-phase fractionation of intact N-linked plasma glycoproteins isolated by lectin affinity columns. The liquid-based fractionation of intact glycoproteins will initially involve nonporous chromatography, followed by liquid capillary isoelectric focusing to further separate protein glycoforms. Each liquid fraction will be spotted on nitrocellulose- coated microscope slides to produce a natural glycoprotein microarray which will then be interrogated by various fluorescently-labeled lectins to probe each microarray spot for the presence of specific glycan moieties. Plasma samples from cancer and normal patients and patients with inflammatory lesions will be analyzed to search for patterns that reveal specific glycan structural changes that occur during cancer progression. Initial analysis will be performed on an analytical test set of 30 samples each from cancer and normal patients and from patients with related inflammatory lesions (chronic pancreatitis) to identify such potential markers. Novel software will be used to analyze these arrays and the resulting test set. Glycoproteins that are associated with cancer versus normal or inflammation will be identified and the structural moieties identified using lectin extraction/LC ESI/MS/MS and QIT-TOF (MALDI MSn) mass spectrometry to examine the detailed changes in structure that may serve as markers of cancer. While many of the marker glycoproteins themselves are of relatively high abundance in plasma, they may be cancer-specific based upon changes in glycan structure. This research has important implications in public health in that it may provide a methodology for searching for markers of disease in plasma. These markers may be related to the stage of the disease and, thus, may become important for personalized treatment, an important issue in clinical medicine. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21CA124441-02
Application #
7467983
Study Section
Enabling Bioanalytical and Biophysical Technologies Study Section (EBT)
Program Officer
Krueger, Karl E
Project Start
2007-07-10
Project End
2011-06-30
Budget Start
2008-07-01
Budget End
2011-06-30
Support Year
2
Fiscal Year
2008
Total Cost
$182,400
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Surgery
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
He, Jintang; Liu, Yashu; Wu, Jing et al. (2013) Analysis of glycoproteins for biomarker discovery. Methods Mol Biol 1002:115-22
Liu, Yashu; He, Jintang; Lubman, David M (2013) Characterization of membrane-associated glycoproteins using lectin affinity chromatography and mass spectrometry. Methods Mol Biol 951:69-77
He, Jintang; Liu, Yashu; Zhu, Thant et al. (2012) CD90 is identified as a candidate marker for cancer stem cells in primary high-grade gliomas using tissue microarrays. Mol Cell Proteomics 11:M111.010744
Li, Chen; Zolotarevsky, Eugene; Thompson, Ian et al. (2011) A multiplexed bead assay for profiling glycosylation patterns on serum protein biomarkers of pancreatic cancer. Electrophoresis 32:2028-35
He, Jintang; Liu, Yashu; Zhu, Thant S et al. (2011) Glycoproteomic analysis of glioblastoma stem cell differentiation. J Proteome Res 10:330-8
Vuong, Huy; Shedden, Kerby; Liu, Yashu et al. (2011) Outlier-Based Differential Expression Analysis in Proteomics Studies. J Proteomics Bioinform 4:116-122
Liu, Yashu; He, Jintang; Li, Chen et al. (2010) Identification and confirmation of biomarkers using an integrated platform for quantitative analysis of glycoproteins and their glycosylations. J Proteome Res 9:798-805
Patwa, Tasneem; Li, Chen; Simeone, Diane M et al. (2010) Glycoprotein analysis using protein microarrays and mass spectrometry. Mass Spectrom Rev 29:830-44
Liu, Yashu; He, Jintang; Xie, Xaiolei et al. (2010) Serum autoantibody profiling using a natural glycoprotein microarray for the prognosis of early melanoma. J Proteome Res 9:6044-51
Feng, Shun; Yang, Na; Pennathur, Subramaniam et al. (2009) Enrichment of glycoproteins using nanoscale chelating concanavalin A monolithic capillary chromatography. Anal Chem 81:3776-83

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