Pulmonary toxicity is a potentially fatal untoward effect of anticancer therapy and our overall goal is to understand the biochemical factors determining drug-induced lung injury. Our central hypothesis is the therapeutic index of the anticancer drug bleomycin (BLM) Can be enhanced with new fundamental information concerning the biochemical basis of BLM- induced lung injury. The major focus, of our proposal will be on the role of bleomycin hydrolase (BH) in protecting lungs against BLM. We also intend to investigate the biological functions of mammalian BH and the factors that regulate its pulmonary expression. We believe these studies will ultimately help to improve the usefulness of BLM by providing therapeutic strategies that reduce cancer treatment morbidity. Damage to the pulmonary endothelium appears to be an early and essential event in fibrosis and we posit that BH levels in the pulmonary endothelium are a critical factor determining pulmonary sensitivity. The full length cDNA for human BH has recently been isolated and sequenced; expression in cells results in BLM resistance. The yeast BH homolog, Gal6, appears to bind DNA making it the first DNA-binding protease. Moreover, the crystal structure has been determined suggesting it is a proteosome-like multimeric enzyme. During the next funding period, we will capitalize on important reagents that have been developed by us and others during the previous four years. We propose to generate transgenic murine models in which BH is either overexpressed or deleted that should permit novel insights into the biological and metabolic role of BH. We expect to test whether predictions from yeast BH concerning structure and function are applicable to human BH by exploring for protein partners and examining protein domains. These studies should assist in understanding the biological and biochemical functions of BH. We will delimit factors that control BH expression.
The Specific Aims are to: 1) develop cells and mice deficient in BH, 2) develop cells and mice that overexpress BH, 3) isolate and characterize potential proteins that interact with BH and, 4) analyze the regulation of BH expression.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
3R01CA043917-13S1
Application #
6096233
Study Section
Experimental Therapeutics Subcommittee 1 (ET)
Project Start
1987-09-01
Project End
2002-01-31
Budget Start
1999-02-01
Budget End
2000-01-31
Support Year
13
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Pittsburgh
Department
Pharmacology
Type
Schools of Medicine
DUNS #
053785812
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Lefterov, I M; Koldamova, R P; Lefterova, M I et al. (2001) Cysteine 73 in bleomycin hydrolase is critical for amyloid precursor protein processing. Biochem Biophys Res Commun 283:994-9
Koldamova, R P; Lefterov, I M; Lefterova, M I et al. (2001) Apolipoprotein A-I directly interacts with amyloid precursor protein and inhibits A beta aggregation and toxicity. Biochemistry 40:3553-60
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Lefterov, I M; Koldamova, R P; Lazo, J S (2000) Human bleomycin hydrolase regulates the secretion of amyloid precursor protein. FASEB J 14:1837-47
Pestell, K E; Ducruet, A P; Wipf, P et al. (2000) Small molecule inhibitors of dual specificity protein phosphatases. Oncogene 19:6607-12
Fabisiak, J P; Pearce, L L; Borisenko, G G et al. (1999) Bifunctional anti/prooxidant potential of metallothionenin: redox signaling of copper binding and release. Antioxid Redox Signal 1:349-64
Schwartz, D R; Homanics, G E; Hoyt, D G et al. (1999) The neutral cysteine protease bleomycin hydrolase is essential for epidermal integrity and bleomycin resistance. Proc Natl Acad Sci U S A 96:4680-5
Koldamova, R P; Lefterov, I M; DiSabella, M T et al. (1999) Human bleomycin hydrolase binds ribosomal proteins. Biochemistry 38:7111-7
Koldamova, R P; Lefterov, I M; Gadjeva, V G et al. (1998) Essential binding and functional domains of human bleomycin hydrolase. Biochemistry 37:2282-90

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