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A comprehensive and innovative proteomics strategy will develop a model investigative approach using novel technologies generally applicable to many NHLBI research interests. Specifically, this Center will employ proteomics-based technologies to investigate protein expression associated with signaling pathways important in inflammatory conditions of the airways.

Specific aims include:

• Develop sensitive, reproducible high-resolution protein separation technologies that function integrally with new high performance imaging and data management software (Nonlinear) to construct protein databases of changes in cellular and subcellular protein expression in the major cell types of inflamed airways. A battery of methodologies, including 1- and 2-dimensional gel methods, subcellular organelle/particle isolation, and novel 2D solution electrophoresis for low abundance proteins will be employed.
• Develop innovative thioaptamer-based array and bead technologies to identify and quantify proteins and protein complexes associated with critical signaling and immune response pathways related to airway inflammation due to respiratory syncytial virus and asthma.
• Utilize specialized high-throughput flow cytometry/cell sorting, technology for laser zapping/imaging for cell selection, magnetic-activated cell sorting, and laser capture microdissection for subsequent proteome analysis of defined cell types in specific stages of the cell cycle.
• Apply systems modeling to correlate changes in protein expression with available genomic data on changes in gene expression due to inflammation after respiratory syncytial virus infection or asthma.
  

Dr. Alexander Kurosky