National Heart, Lung, and Blood Institute Proteomics Initiative

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SOLICITATION NUMBER: NHLBI-HL-02-04, CONTRACT NUMBER: NO1-HV-28187 OFFEROR NAME AND ADDRESS: Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc. 1401 Rockville Pike, Suite 600 Rockville MD 20852 RFP TITLE: National Heart, Lung, and Blood Proteomics Initiative PRINCIPAL INVESTIGATOR: Dr. Harvey B. Pollard, M.D., Ph.D. 3D Proteomics in Cystic Fibrosis SUMMARY OF OBJECTIVES: The Center for Proteomics Research is a highly interactive, multidisciplinary program, designed to develop innovative proteomic technologies and apply them to the NHLBI-relevant disease of Cystic Fibrosis (CF). The goal of the present project is to deliver 3-D Proteomics, an innovative, sensitive, quantitative, and dynamic technology for enumerating the complete CF proteome from CF lung cells and CF patient biopsy tissue. The novelty in this proposal is the 3rd dimension a quantitative, biosynthetic rate for each protein in the proteome. To achieve this deliverable we propose the following: Specific Objective #1: To identify the proteins in cultured CF lung epithelial cells which are affected by CF, using the technique of 3-D proteomics. This technology involves pulsing lines of CF and isogenic control cells with 35[S] methionine or cysteine, separating proteins on 2-D gels, and detecting proteins by radioimaging. Specific Objective #2: To identify all proteins in patient samples of CF lung epithelial cells which areaffected by the disease, using 3-D Proteomics. Epithelial cells from biopsies of CF lungs will bepulse labeled with 35[S] methionine or cysteine. Significance: 3-D Proteomics, offers a rapid, sensitive and stable real time surrogate endpoint for both CF diagnosis and patient monitoring at the bedside, and a platform for discovery of novel CF therapeutics at the bench.