Application of the adductome approach to assess intertissue DNA damage variations in human lung and esophagus

Abstract

Methods for determining the differential susceptibility of human organs to DNA damage have not yet been explored to any large extent due to technical constraints. The development of comprehensive analytical approaches by which to detect intertissue variations in DNA damage susceptibility may advance our understanding of the roles of DNA adducts in cancer etiology and as exposure biomarkers at least. A strategy designed for the detection and comparison of multiple DNA adducts from different tissue samples was applied to assess esophageal and peripherally- and centrally-located lung tissue DNA obtained from the same person. This adductome approach utilized LC/ESI-MS/MS analysis methods designed to detect the neutral loss of 2′-deoxyribose from positively ionized 2′-deoxynucleoside adducts transmitting the [M+H] + > [M+H−116] + transition over 374 transitions. In the final analyses, adductome maps were produced which facilitated the visualization of putative DNA adducts and their relative levels of occurrence and allowed for comprehensive comparisons between samples, including a calf thymus DNA negative control. The largest putative adducts were distributed similarly across the samples, however, differences in the relative amounts of putative adducts in lung and esophagus tissue were also revealed. The largest-occurring lung tissue DNA putative adducts were 90% similar ( n = 50), while putative adducts in esophagus tissue DNA were shown to be 80 and 84% similar to central and peripheral lung tissue DNA respectively. Seven DNA adducts, N 2-ethyl-2′-deoxyguanosine ( N 2-ethyl-dG), 1, N 6-etheno-2′-deoxyadenosine (ɛdA), α- S- and α- R-methyl-γ-hydroxy-1, N 2-propano-2′-deoxyguanosine (1, N 2-PdG 1, 1, N 2-PdG 2), 3-(2′-deoxyribosyl)-5,6,7,8-tetrahydro-8-hydroxy-pyrimido[1,2- a]purine-(3 H)-one (8-OH-PdG) and the two stereoisomers of 3-(2′-deoxyribosyl)-5,6,7,8-tetrahydro-6-hydroxypyrimido[1,2- a]purine-(3 H)-one (6-OH-PdG) were unambiguously detected in all tissue DNA samples by comparison to authentic adduct standards and stable isotope dilution and their identities were matched to putative adducts detected in the adductome maps.