Abstract

As much as 60% of all renal disease may involve a nephrotoxin. Mercapturic acid biosynthesis is an important xenobiotic detoxiciation system in mammals. The kidney plays a central role in the processing of mercapturate pathway intermediates and the excretion of the mercapturic acid. A variety of cysteine conjugates formed via the mercapturate pathway are nephrotoxic. The mechanism for toxicity of the conjugates is unclear but may involve activation by cysteine conjugate beta-lyase. However, recent data suggest that other pathways may be involved and the mechanism of toxicity may differ depending on the conjugate. Rat kidney cells in culture are a good model to study the mechanism of cysteine conjugate toxicity. Understanding the structure activity relationship for toxicity and metabolism is an important key to understanding the mechanism. Dose response relationships will be established for toxicity. Effects on cell function will be evaluated. The role of the cysteine conjugate beta-lyase pathway will be tested. The cysteine conjugate beta- lyases will be purified and the structure activity for metabolism will be determined. New pathways of cysteine conjugate metabolism will be studies and the toxicity of novel metabolites determined. Radiolabelled conjugates will be synthesized and covalent binding measured in cells and with a model protein. Selective blocking agents which block binding but do not influence metabolism will be used to investigate the role of covalent binding in toxicity. Cellular targets will be identified using the culture system. The data will be used to predict the toxicity and to study the cell biology of nephrotoxic damage and repair.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK038925-05
Application #
3238511
Study Section
Toxicology Subcommittee 2 (TOX)
Project Start
1988-04-01
Project End
1993-03-31
Budget Start
1992-04-01
Budget End
1993-03-31
Support Year
5
Fiscal Year
1992
Total Cost
Indirect Cost

  Institution

Name
Adirondack Biomedical Research Institute
Department
Type
DUNS #
City
Lake Placid
State
NY
Country
United States
Zip Code
12946

  Publications

Chen, Q; Jones, T W; Stevens, J L (1994) Early cellular events couple covalent binding of reactive metabolites to cell killing by nephrotoxic cysteine conjugates. J Cell Physiol 161:293-302
Yu, K; Chen, Q; Liu, H et al. (1994) Signalling the molecular stress response to nephrotoxic and mutagenic cysteine conjugates: differential roles for protein synthesis and calcium in the induction of c-fos and c-myc mRNA in LLC-PK1 cells. J Cell Physiol 161:303-11
Zhang, G; Ichimura, T; Maier, J A et al. (1993) A role for fibroblast growth factor type-1 in nephrogenic repair. Autocrine expression in rat kidney proximal tubule epithelial cells in vitro and in the regenerating epithelium following nephrotoxic damage by S-(1,1,2,2-tetrafluoroethyl)-L-cysteine in vi J Biol Chem 268:11542-7
Bruschi, S A; West, K A; Crabb, J W et al. (1993) Mitochondrial HSP60 (P1 protein) and a HSP70-like protein (mortalin) are major targets for modification during S-(1,1,2,2-tetrafluoroethyl)-L-cysteine-induced nephrotoxicity. J Biol Chem 268:23157-61
Fisher, M B; Hayden, P J; Bruschi, S A et al. (1993) Formation, characterization, and immunoreactivity of lysine thioamide adducts from fluorinated nephrotoxic cysteine conjugates in vitro and in vivo. Chem Res Toxicol 6:223-30
Chen, Q; Yu, K; Stevens, J L (1992) Regulation of the cellular stress response by reactive electrophiles. The role of covalent binding and cellular thiols in transcriptional activation of the 70-kilodalton heat shock protein gene by nephrotoxic cysteine conjugates. J Biol Chem 267:24322-7
Hayden, P J; Welsh, C J; Yang, Y et al. (1992) Formation of mitochondrial phospholipid adducts by nephrotoxic cysteine conjugate metabolites. Chem Res Toxicol 5:232-7
Wallin, A; Zhang, G; Jones, T W et al. (1992) Mechanism of the nephrogenic repair response. Studies on proliferation and vimentin expression after 35S-1,2-dichlorovinyl-L-cysteine nephrotoxicity in vivo and in cultured proximal tubule epithelial cells. Lab Invest 66:474-84
Chen, Q; Yu, K; Holbrook, N J et al. (1992) Activation of the growth arrest and DNA damage-inducible gene gadd 153 by nephrotoxic cysteine conjugates and dithiothreitol. J Biol Chem 267:8207-12
Chen, Q; Stevens, J L (1991) Inhibition of iodoacetamide and t-butylhydroperoxide toxicity in LLC-PK1 cells by antioxidants: a role for lipid peroxidation in alkylation induced cytotoxicity. Arch Biochem Biophys 284:422-30

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