Acute overdosage with the antipyretic analgesic, acetaminophen (APAP) can cause severe, fatal hepatic necrosis. Hepatotoxicity is the result of APAP activation to an electrophile which covalently binds to liver proteins. Although, the relationship between APAP's protein covalent binding and toxicity is not clear, selective arylation of a 58 kDa APAP binding protein (58-ABP) is better associated with toxicity than is total covalent binding. The 58-ABP is the most prominent target detected with anti-APAP antibody in mouse and human liver during APAP hepatotoxicity. In mice, 58-ABP arylation was most closely associated with target tissue damage (liver, lung and kidney) in varied experimental paradigms. The arylated protein is also detected in mouse plasma after toxic APAP exposures, and this may mirror the detection of APAP-protein adducts in human plasma during poisoning. Our working hypothesis is that arylation of the 58-ABP is important for APAP-induced target organ toxicity. Whether is plays an initiating or protective role cannot presently be resolved. Recent results have raised new questions which are the basis of the proposed research. 1) Is there a threshold for 58-ABP binding which, if exceeded, is associated with toxicity? 2) Is the 58-ABP also targeted by other agents which bind covalently to proteins? 3) What is the toxicological significance of the appearance of arylated 58-ABP in plasma? The proposed research will extensively use anti-APAP and anti-58, a new antibody prepared against the 58-ABP. This will permit quantitative assessment of the existence of a covalent binding threshold. Antibodies will be developed against bound bromobenzene to permit comparison of its binding with APAP's and with bromobenzene toxicity. Immunoprecipitation with anti-58 will facilitate detection of 58-ABP binding by other xenobiotics. In vitro studies will determine if other agents bind at the same site on the 58-ABP and APAP, and culture and in vivo studies will reveal if such binding can cause toxic interactions during simultaneous exposures. We will also determine if the appearance of 58-ABP in plasma after APAP or other xenobiotics reflects cellular secretion of arylated protein or incidental loss as a result of cell death. Studies with agents which bind but do not cause toxicity will also be included to determine if such agents target the same proteins as the toxic compounds. Collectively such studies will provide new insight into the importance of selective 58-ABP arylation by APAP and other toxic chemicals for target organ toxicity. They will also determine if the appearance of 58-ABP in plasma can become a useful diagnostic marker for xenobiotic-induced target organ damage.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM031460-08
Application #
3279464
Study Section
Toxicology Subcommittee 2 (TOX)
Project Start
1984-08-01
Project End
1995-11-30
Budget Start
1992-12-01
Budget End
1993-11-30
Support Year
8
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of Connecticut
Department
Type
Schools of Pharmacy
DUNS #
City
Storrs-Mansfield
State
CT
Country
United States
Zip Code
06269
Slitt, Angela M Lucas; Dominick, Pamela K; Roberts, Jeanette C et al. (2005) Effect of ribose cysteine pretreatment on hepatic and renal acetaminophen metabolite formation and glutathione depletion. Basic Clin Pharmacol Toxicol 96:487-94
Slitt, Angela Lucas; Dominick, Pamela K; Roberts, Jeanette C et al. (2004) Standard of care may not protect against acetaminophen-induced nephrotoxicity. Basic Clin Pharmacol Toxicol 95:247-8
Lucas, A M; Hennig, G; Dominick, P K et al. (2000) Ribose cysteine protects against acetaminophen-induced hepatic and renal toxicity. Toxicol Pathol 28:697-704
Boulares, A H; Giardina, C; Inan, M S et al. (2000) Acetaminophen inhibits NF-kappaB activation by interfering with the oxidant signal in murine Hepa 1-6 cells. Toxicol Sci 55:370-5
Boulares, H A; Giardina, C; Navarro, C L et al. (1999) Modulation of serum growth factor signal transduction in Hepa 1-6 cells by acetaminophen: an inhibition of c-myc expression, NF-kappaB activation, and Raf-1 kinase activity. Toxicol Sci 48:264-74
Boess, F; Bopst, M; Althaus, R et al. (1998) Acetaminophen hepatotoxicity in tumor necrosis factor/lymphotoxin-alpha gene knockout mice. Hepatology 27:1021-9
Cohen, S D; Khairallah, E A (1997) Selective protein arylation and acetaminophen-induced hepatotoxicity. Drug Metab Rev 29:59-77
Cohen, S D; Pumford, N R; Khairallah, E A et al. (1997) Selective protein covalent binding and target organ toxicity. Toxicol Appl Pharmacol 143:1-12
Navarro, C L; Cohen, S D; Khairallah, E A (1996) Genes encoding the acetaminophen and selenium binding proteins map to mouse chromosome 3. Mamm Genome 7:919-20
Emeigh Hart, S G; Wyand, D S; Khairallah, E A et al. (1996) Acetaminophen nephrotoxicity in the CD-1 mouse. II. Protection by probenecid and AT-125 without diminution of renal covalent binding. Toxicol Appl Pharmacol 136:161-9

Showing the most recent 10 out of 43 publications