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Grant Details

Grant Number: 5R21CA140036-03 Interpret this number
Primary Investigator: Decaprio, Anthony
Organization: Florida International University
Project Title: Platform for High-Throughput Analysis of Protein Adducts for Carcinogen Exposure
Fiscal Year: 2013


DESCRIPTION (provided by applicant): The long-term goal of this project is to develop a high-throughput technology that can characterize covalent protein and DNA modifications ("adducts") induced by reactive carcinogens on a global basis with high sensitivity and specificity. Virtually all carcinogens, even the most inert, either contain an electrophilic center or undergo some level of oxidative metabolism to form reactive electrophiles. DNA and protein adducts are the products of the chemical reaction of such electrophiles with nucleophilic sites within these macromolecules. DNA adduction (including methylation and oxidative changes) has been widely implicated in mutation and carcinogenesis, while protein adducts (i.e., post-translational modifications) may have indirect roles in the mechanism of carcinogenesis if they involve DNA-related proteins or increase the rate of tumor promotion. In addition, such adducts are being explored as short- term, long-term, and/or cumulative markers of exposure to carcinogens. Current methods of adduct quantitation are expensive, highly chemical-specific, and labor-intensive. A technology for rapid and comprehensive profiling of macromolecular adduction in human subjects would be an important tool for carcinogen exposure and health risk assessment. This exploratory/developmental application proposes a proof-of-concept study to adapt three recently developed technologies, i.e., combinatorial chemical synthesis, antibody phage display, and microarray technology in a system to rapidly profile blood protein adducts of two important classes of carcinogens. Libraries of adducted peptides will be created to mimic structures formed in vivo in humans exposed to carcinogens. Adduct-specific scFv probes will be selected by screening with phage display libraries and validated with human blood specimens in an ELISA system. Results of ELISA-based adduct quantitation will be compared to those from conventional mass spectrometry-based analysis of blood protein adducts. If the proposed proof-of-concept studies are successful, expanded funding will be sought to develop a microarray-based global adduct detector for screening blood specimens from individual subjects. It is anticipated that the technology developed in this project will have wide applications for carcinogen exposure assessment, biomarker discovery, clinical diagnosis, and assessment of cancer treatment efficacies.


Proteomic Analysis of Thiol Modifications and Assessment of Structural Changes in Hemoglobin Induced by the Aniline Metabolites N-Phenylhydroxylamine and Nitrosobenzene.
Authors: Möller C. , Davis W.C. , Thompson V.R. , Marí F. , DeCaprio A.P. .
Source: Scientific reports, 2017-11-01; 7(1), p. 14794.
EPub date: 2017-11-01.
PMID: 29093547
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