Mast cell cysteine, metalloprotease, and serine proteases may play a significant role in the pathogenesis of inflammatory lung diseases. The long-term objective of this proposal is to identify the cysteine proteases present in mast cells, and to characterize their roles in mast cell dependent inflammation. We hypothesize that tissue mast cells are a major source of dipeptidyl peptidase I (DPPI) and a newly identified 60-kDa cysteine protease and that these proteases participate in inflammation by hydrolyzing extracellular proteins.
Specific aims are : 1. To explore structure-function relationships of DPPI; 2. To identify cells expressing DPPI in normal and inflamed lung tissues; 3. To characterize substrate specificity of DPPI, and 4. To characterize a nove1 60 kDa cysteine protease found in mast cells. To achieve these goals: 1. Proteolytic processing of proDPPI will be studied in recombinantly expressed DPPI. 2. DPPI's expressed with mutated propeptides will be assayed for their dipeptidyl peptidase and endoprotease activity, 3. Expression of DPPI by specific cell types in normal and inflamed dog lungs will be determined by immunohistochemistry and in situ hybridization. 4. DPPI will be studied for its ability to hydrolyze extracellular peptides and proteins. 5. Mast cell functional characteristics and proteolytic activities will be studied in DPPI knockout mice. 6. A novel mast cell cysteine protease will be purified and characterized. Dr. Wolters has completed training in internal medicine and pulmonary and critical care medicine. He has demonstrated a firm commitment to a career in academic medicine and an interest in studying the pathophysiology of lung diseases. The research training plan includes: an intensive laboratory experience, didactic coursework, weekly seminars, and journal clubs. The research training will be overseen by an advisory committee including the sponsor (Dr. Caughey) and experts on cysteine proteases (Dr. McKerrow), immunohistopathology (Dr. McDonald), and pulmonary diseases (Dr. Matthay), who will guide the candidates development into an independent investigator in pulmonary research.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Clinical Investigator Award (CIA) (K08)
Project #
1K08HL004055-01
Application #
2881397
Study Section
Special Emphasis Panel (ZHL1-CSR-K (M1))
Project Start
1999-08-04
Project End
2004-07-31
Budget Start
1999-08-04
Budget End
2000-07-31
Support Year
1
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of California San Francisco
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Kim, Kevin K; Kugler, Matthias C; Wolters, Paul J et al. (2006) Alveolar epithelial cell mesenchymal transition develops in vivo during pulmonary fibrosis and is regulated by the extracellular matrix. Proc Natl Acad Sci U S A 103:13180-5
Wolters, P J; Mallen-St Clair, J; Lewis, C C et al. (2005) Tissue-selective mast cell reconstitution and differential lung gene expression in mast cell-deficient Kit(W-sh)/Kit(W-sh) sash mice. Clin Exp Allergy 35:82-8
Mallen-St Clair, Jon; Pham, Christine T N; Villalta, S Armando et al. (2004) Mast cell dipeptidyl peptidase I mediates survival from sepsis. J Clin Invest 113:628-34
Henningsson, Frida; Wolters, Paul; Chapman, Harold A et al. (2003) Mast cell cathepsins C and S control levels of carboxypeptidase A and the chymase, mouse mast cell protease 5. Biol Chem 384:1527-31
Shi, G-P; Sukhova, G K; Kuzuya, M et al. (2003) Deficiency of the cysteine protease cathepsin S impairs microvessel growth. Circ Res 92:493-500
Muilenburg, Diego J; Raymond, Wilfred W; Wolters, Paul J et al. (2002) Lys40 but not Arg143 influences selectivity of angiotensin conversion by human alpha-chymase. Biochim Biophys Acta 1596:346-56
Wolters, P J; Pham, C T; Muilenburg, D J et al. (2001) Dipeptidyl peptidase I is essential for activation of mast cell chymases, but not tryptases, in mice. J Biol Chem 276:18551-6
Caughey, G H; Raymond, W W; Wolters, P J (2000) Angiotensin II generation by mast cell alpha- and beta-chymases. Biochim Biophys Acta 1480:245-57
Caughey, G H; Raymond, W W; Blount, J L et al. (2000) Characterization of human gamma-tryptases, novel members of the chromosome 16p mast cell tryptase and prostasin gene families. J Immunol 164:6566-75
Wolters, P J; Chapman, H A (2000) Importance of lysosomal cysteine proteases in lung disease. Respir Res 1:170-7