Mass spectrometry is an extraordinarily powerful bio-analytical technique that has had a profound impact on our molecular understanding of human health and disease. Major advances in mass analyzer technology, dissociation techniques, and ionization methods are largely attributed to the central role that mass spectrometry plays in the field of systems biology. While mass spectrometry has evolved over the last century into a highly effective analytical tool, there are still opportunities for new advances to be made allowing an even more diverse array of biological questions to be addressed. This proposal is centered on the development and characterization of novel glycan- specific tags for biological mass spectrometry that facilitate the relative quantification of glycans cleaved from plasma proteins with increased ion abundance. The short-term objective of this proposal is to develop these novel tags and then test them using the avian model of spontaneous epithelial ovarian cancer. These results will provide a solid bio-analytical technology foundation from which our newly developed chemical tags can be applied to achieve the long-term objective of effectively elucidating glycan biomarkers for the early detection of epithelial ovarian cancer in women. Public Description of Proposed Research Mass spectrometry (MS), the science related to the """"""""weighing of molecules"""""""", has had a profound impact on the study of human health and disease including cancer, heart disease, neural development, and auto-immune diseases. However, front-end chemistries for MS-based glycomics will be exploited to dramatically improve the ability of MS to detect and quantify glycans cleaved from plasma proteins. This will allow a more diverse array of contemporary biomedical questions to be addressed including the quantification of diagnostic and prognostic biomarkers. This proposal is squarely centered on the elucidation of a glycan-specific biomarker(s) for the early detection of epithelial ovarian cancer.

Public Health Relevance

This proposal seeks support to develop and apply novel tags that will facilitate both global and targeted quantitative mass spectrometric analysis of N-linked glycans with significantly improved limits-of-detection. Once synthesized, the novel tags and ancillary methods will concurrently be applied to the experimental chicken model of spontaneous epithelial ovarian cancer for comparative analysis (i.e., biomarker discovery) and fully disseminated to the glycomics research community. The long-term application of these novel tags, as developed in our laboratory for this study, will be their use to elucidate glycan biomarkers for the early detection of epithelial ovarian cancer.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants Phase II (R33)
Project #
5R33CA147988-02
Application #
8327164
Study Section
Special Emphasis Panel (ZCA1-SRLB-X (J1))
Program Officer
Patriotis, Christos F
Project Start
2011-09-01
Project End
2014-08-31
Budget Start
2012-09-01
Budget End
2013-08-31
Support Year
2
Fiscal Year
2012
Total Cost
$353,783
Indirect Cost
$112,874
Name
North Carolina State University Raleigh
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
042092122
City
Raleigh
State
NC
Country
United States
Zip Code
27695
Hecht, Elizabeth S; McCord, James P; Muddiman, David C (2016) A Quantitative Glycomics and Proteomics Combined Purification Strategy. J Vis Exp :
Hecht, Elizabeth S; Scholl, Elizabeth H; Walker, S Hunter et al. (2015) Relative Quantification and Higher-Order Modeling of the Plasma Glycan Cancer Burden Ratio in Ovarian Cancer Case-Control Samples. J Proteome Res 14:4394-401
Hecht, Elizabeth S; McCord, James P; Muddiman, David C (2015) Definitive Screening Design Optimization of Mass Spectrometry Parameters for Sensitive Comparison of Filter and Solid Phase Extraction Purified, INLIGHT Plasma N-Glycans. Anal Chem 87:7305-12
Randall, Shan M; Koryakina, Irina; Williams, Gavin J et al. (2014) Evaluating nonpolar surface area and liquid chromatography/mass spectrometry response: an application for site occupancy measurements for enzyme intermediates in polyketide biosynthesis. Rapid Commun Mass Spectrom 28:2511-22
Walker, S Hunter; Taylor, Amber D; Muddiman, David C (2013) The use of a xylosylated plant glycoprotein as an internal standard accounting for N-linked glycan cleavage and sample preparation variability. Rapid Commun Mass Spectrom 27:1354-8
Walker, S Hunter; Taylor, Amber D; Muddiman, David C (2013) Individuality Normalization when Labeling with Isotopic Glycan Hydrazide Tags (INLIGHT): a novel glycan-relative quantification strategy. J Am Soc Mass Spectrom 24:1376-84
Walker, S Hunter; Carlisle, Brandon C; Muddiman, David C (2012) Systematic comparison of reverse phase and hydrophilic interaction liquid chromatography platforms for the analysis of N-linked glycans. Anal Chem 84:8198-206
Walker, S Hunter; Budhathoki-Uprety, Januka; Novak, Bruce M et al. (2011) Stable-isotope labeled hydrophobic hydrazide reagents for the relative quantification of N-linked glycans by electrospray ionization mass spectrometry. Anal Chem 83:6738-45
Walker, S Hunter; Lilley, Laura M; Enamorado, Monica F et al. (2011) Hydrophobic derivatization of N-linked glycans for increased ion abundance in electrospray ionization mass spectrometry. J Am Soc Mass Spectrom 22:1309-17
Barry, Jeremy A; Muddiman, David C (2011) Global optimization of the infrared matrix-assisted laser desorption electrospray ionization (IR MALDESI) source for mass spectrometry using statistical design of experiments. Rapid Commun Mass Spectrom 25:3527-36

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