Hexavalent chromium (Cr-6) is a firmly established human carcinogen with widespread exposure in metal- processing and other industries. There are also significant concerns about potential health effects of environmental exposure due to the presence of Cr-6 at numerous Superfund sites and its ambient and drinking water contamination. Cr-6 is a pro-carcinogen that is reduced inside the cell yielding stable Cr-3 and causing the formation of abundant Cr-DNA adducts. Cr-DNA adducts are mutagenic and genotoxic in human cells although they are weakly duplex-distorting and do not block DNA replication in vitro. We have found that genotoxicity of Cr-DNA adducts in human or mouse cells required the presence of mismatch repair (MMR) proteins. Cells lacking MMR activity were highly resistant to the induction of cell death and chromosomal damage by Cr-6. Chronic exposure to Cr-6 can therefore lead to the selection of resistant cells with inactive MMR, which provides an explanation for the very high frequency of microsatellite instability (a hallmark of inactive MMR) among lung cancers in chromate workers. Our recent findings also showed that MMR enhances Cr-6 genotoxicity via a rapid production of DNA double-strand breaks (DSB) in G2 cells immediately after replication of Cr-damaged DNA. The induction of DSB required a sequential recruitment of MSH6- and MSH3-containing complexes, indicating the unprecedented cooperation of both MMR branches in processing of Cr-DNA adducts. This application is designed to address three central questions related to the formation and the biological importance of MMR-induced DSB. These include: (1) mechanisms of DSB generation and the role of individual MMR proteins, (2) significance of error-prone and error-free pathways in repair of MMR-induced DSB, and (3) crosstalk between DSB processing and checkpoint signaling. The completion of the proposed work should identify the key processes involved in the formation and repair of highly toxic DNA lesions responsible for chromosomal breakage and other adverse genetic effects resulting from human exposure to Cr-6. Elucidation of molecular mechanisms involved in the unique processing of Cr-damaged DNA is also expected to uncover novel functions of MMR, a key genome maintenance system and a major factor in the clinical efficacy of platinum-based and some other DNA alkylating chemotherapeutic drugs.

Public Health Relevance

Hexavalent chromium is a potent human carcinogen with widespread environmental and occupational exposures. This toxic metal causes DNA breaks that remain in cells for a long period of time leading to adverse genetic effects even at very low chromium doses. This proposal is focused on the determination of the molecular causes of DNA breaks in chromium-exposed human cells and the characterization of the unique cellular systems required for their repair. The completion of the proposed work can lead to the identification of biochemical factors that increase or decrease individual susceptibility to genetic damage by carcinogenic chromium.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES008786-15
Application #
8257147
Study Section
Special Emphasis Panel (ZRG1-DKUS-B (03))
Program Officer
Reinlib, Leslie J
Project Start
1997-08-01
Project End
2014-04-30
Budget Start
2012-05-01
Budget End
2013-04-30
Support Year
15
Fiscal Year
2012
Total Cost
$357,247
Indirect Cost
$136,724
Name
Brown University
Department
Pathology
Type
Schools of Medicine
DUNS #
001785542
City
Providence
State
RI
Country
United States
Zip Code
02912
DeLoughery, Zachary; Luczak, Michal W; Zhitkovich, Anatoly (2014) Monitoring Cr intermediates and reactive oxygen species with fluorescent probes during chromate reduction. Chem Res Toxicol 27:843-51
Wong, Victor C; Morse, Jessica L; Zhitkovich, Anatoly (2013) p53 activation by Ni(II) is a HIF-1* independent response causing caspases 9/3-mediated apoptosis in human lung cells. Toxicol Appl Pharmacol 269:233-9
Luczak, Michal W; Zhitkovich, Anatoly (2013) Role of direct reactivity with metals in chemoprotection by N-acetylcysteine against chromium(VI), cadmium(II), and cobalt(II). Free Radic Biol Med 65:262-9
Morse, Jessica L; Luczak, Michal W; Zhitkovich, Anatoly (2013) Chromium(VI) causes interstrand DNA cross-linking in vitro but shows no hypersensitivity in cross-link repair-deficient human cells. Chem Res Toxicol 26:1591-8
Zhitkovich, Anatoly (2011) Chromium in drinking water: sources, metabolism, and cancer risks. Chem Res Toxicol 24:1617-29
Zecevic, Alma; Hagan, Elizabeth; Reynolds, Mindy et al. (2010) XPA impacts formation but not proteasome-sensitive repair of DNA-protein cross-links induced by chromate. Mutagenesis 25:381-8
Macfie, Andrea; Hagan, Elizabeth; Zhitkovich, Anatoly (2010) Mechanism of DNA-protein cross-linking by chromium. Chem Res Toxicol 23:341-7
Zecevic, Alma; Menard, Haley; Gurel, Volkan et al. (2009) WRN helicase promotes repair of DNA double-strand breaks caused by aberrant mismatch repair of chromium-DNA adducts. Cell Cycle 8:2769-78
Reynolds, Mindy F; Peterson-Roth, Elizabeth C; Bespalov, Ivan A et al. (2009) Rapid DNA double-strand breaks resulting from processing of Cr-DNA cross-links by both MutS dimers. Cancer Res 69:1071-9
Guttmann, David; Poage, Graham; Johnston, Tatiana et al. (2008) Reduction with glutathione is a weakly mutagenic pathway in chromium(VI) metabolism. Chem Res Toxicol 21:2188-94

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