Evidence for Covalent DNA Adduction by Ochratoxin A  

1: Chem Res Toxicol. 2004 Sep 20;17(9):1289-1296.

Evidence for Covalent DNA Adduction by Ochratoxin A following Chronic Exposure to Rat and Subacute Exposure to Pig

Faucet V, Pfohl-Leszkowicz A, Dai J, Castegnaro M, Manderville RA.

Department Toxicology & Food Safety, Lab. Genie Chimique, Ecole Nationale Superieure Agronomique de Toulouse, UMR-CNRS 5503, Agrobiopole-BP 107, F31326 Castanet, Tolosan Cedex, France, Department of Chemistry, Wake Forest University, Winston-Salem, North Carolina 27109-7486, IARC, 150 Cours Albert Thomas, 69000 Lyon, France, and Department of Chemistry, University of Guelph, Guelph, Ontario N1G 2W1, Canada.

Ochratoxin A (OTA) is a nephrotoxic mycotoxin that is a potent renal carcinogen in male rats and is suspected of being the etiological agent of Balkan endemic nephropathy (BEN) and its associated urinary tract cancers. Conflicting results have been obtained regarding the genotoxicity of OTA and its ability to react directly with DNA upon oxidative bioactivation to yield covalent DNA adducts. To characterize DNA adduction by OTA, the present study utilizes the photooxidative properties of the toxin to generate authentic C8 OTA-3'-monophosphate-deoxyguanosine (3'-dGMP) adducts for use as cochromatographic standards for (32)P-postlabeling detection of OTA-mediated DNA adduction in the kidney of rat and pig. Our results show evidence for the photooxidation of OTA to yield carbon (C)- and oxygen (O)-bonded C8-3'-dGMP adducts (C-C8 and O-C8) that have been isolated and characterized by LC/MS with in-line UV and electrospray negative ionization (ES(-)) detection. A comparison to previously published work on related C8-dG adducts supports C8 attachment by OTA. The C-C8 OTA-3'-dGMP adduct standard is shown by (32)P-postlabeling to comigrate with the major lesion detected in the kidney of rat following chronic exposure to OTA and with one of four adducts detected in the kidney of pig following subacute exposure to the toxin. The O-C8 OTA-3'-dGMP adduct standard is also shown to coelute with a lesion detected in rat kidney. These findings suggest a role for the OTA phenoxyl radical in OTA-mediated DNA adduction in vivo, provide a rationale for the tumorigenesis of OTA, and strengthen the OTA hypothesis in the etiology of BEN and the associated urinary tract tumors.