Colorectal cancer (CRC) is one of the most common cancers in the United States and the second ranked cause of cancer related death. This project proposes to perform both broad proteomic and glycomic screens and targeted analyses to discover early detection and diagnostic biomarkers of CRC. The goal is to improve colon screening by the addition of highly sensitive markers that together with each other or existing markers yield good specificity. We will interrogate unique prediagnostic plasma samples, tissue from primary CRC tumors, and plasma from CRC cases on custom high-density antibody microarrays (up to 6000 different analytes) to discover novel proteomic and glycomic biomarkers. Parallel arrays will be incubated with plasma or tissue lysate for proteomic comparison or glycosylation (via incubation with different lectins). This microarray approach employs technology with unparalleled sensitivity, enabling interrogation down to the low picomolar concentration of the circulating proteome. Biomarker candidates that pass the statistical threshold on a "discovery" array will be retained and be re-examined using new samples on smaller arrays (to reduce the false discovery rate) - a step we will refer to as "pre-validation" to indicate that a potential biomarker would have shown up as statistically significant in multiple sample sets but was not yet subjected to formal characterization or validation. This unique triage process allows for rapid sorting of potential biomarkers, allowing us to focus on only the most promising. The best candidates will then be evaluated with more conventional ELISA and lectin based methods. Our approaches will specifically target proteomic and glycomic changes in proteins critical for the regulation of apoptosis, proliferation, angiogenesis, inflammation/prostaglandins, insulin resistance, toll-like receptor (TLR), transforming growth factor (TGF)-? and STAT signaling pathway deregulation during CRC to discover biomarkers of early detection and diagnosis. These include over 300 phospho-specific antibodies and their matched non-phospho-specific counterparts to 21 MAPK, 12 TGF-?, and many STAT pathway targets.

Agency
National Institute of Health (NIH)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01CA152746-05
Application #
8686771
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Rinaudo, Jo Ann S
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
City
Seattle
State
WA
Country
United States
Zip Code
98109
Grady, William M; Pritchard, Colin C (2014) Molecular alterations and biomarkers in colorectal cancer. Toxicol Pathol 42:124-39
Rho, Jung-hyun; Mead, Judson R; Wright, W Shea et al. (2014) Discovery of sialyl Lewis A and Lewis X modified protein cancer biomarkers using high density antibody arrays. J Proteomics 96:291-9
Rho, Jung-hyun; Lampe, Paul D (2013) High-throughput screening for native autoantigen-autoantibody complexes using antibody microarrays. J Proteome Res 12:2311-20