Dissertation/Thesis Abstract

CEdG – a glycated DNA adduct linking altered metabolism and genetic instability
by Tamae, Daniel, Ph.D., City of Hope's Irell & Manella Graduate School of Biomedical Sciences, 2011, 213; 3596728
Abstract (Summary)

This dissertation details original work focused on the DNA adduct N2-(1-carboxyethyl)-2'-deoxyguanosine (CEdG). This DNA adduct results from the spontaneous reaction of DNA with the endogenous and exogenous formed, carbohydrate-derived, reactive carbonyl species, methylglyoxal. Using in vitro steady state kinetics, we have shown that CEdG in template DNA leads to DNA miscoding effects when the model replicative polymerase, exonuclease-free Klenow fragment (KF-) is used. The development, validation and application of a novel stable isotope dilution, triple quadrupole mass spectrometric method for the quantitation of CEdG is also detailed. This method was used to quantitate CEdG in urine from diabetic rats, urine from human patients, human tumor and adjacent biopsy tissue, diabetic animal tissue and DNA treated with methylglyoxal. Finally, we detail the adaptation, validation and application of a novel, commercially-available microfluidic HPLC-chip for increased sensitivity in the quantitation of CEdG and also apply it to the quantitation of the RNA analogue, CEG. Combined, these studies establish CEdG as a potential biomarker for glycation and point to a viable avenue for connecting chronic glycolytic flux with genetic instability.

Indexing (document details)
Advisor: Termini, John
Commitee: Bailis, Adam, O'Connor, Timothy, Sowers, Lawrence, Synold, Timothy
School: City of Hope's Irell & Manella Graduate School of Biomedical Sciences
Department: Biological Sciences
School Location: United States -- California
Source: DAI-B 75/01(E), Dissertation Abstracts International
Subjects: Analytical chemistry, Biochemistry
Keywords: Advanced glycation end products, Cancer, DNA adducts, Diabetes, Mass spectrometry
Publication Number: 3596728
ISBN: 9781303439254
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