Abstract: The tools of modern biology have revolutionized biomedical research, enabling an exponential growth in the acquisition of data regarding genes, proteins, and their structures and functions in normal and diseased states. Among these advances, the ability to monitor profiles of genes and protein expression accurately and on a large scale is notable. However, it is often not easy to correlate the trends and relationships observed in normal or abnormal states to the phenotype resulting from the gene expression profile. We are developing a new functional genomics approach for studying gene expression dynamics involving simultaneous measurement of temporal expression profiles of multiple genes. The technique relies on live cell imaging of fluorescence reporter cells in a microfabricated fluidically addressable cell array. This high throughput format allows parallel control of diverse soluble stimuli (concentrations and dynamics of cytokines, growth factors, and potential therapeutics) while allowing simultaneous noninvasive monitoring of gene expression at a single cell level. In this work our goals are to: 1) obtain the genes whose expression levels are altered by molecular mediators of the stress response and to generate green fluorescence protein (GFP)-tagged expression constructs of these genes; 2) use microfabrication and microfluidic techniques for developing living cell microarrays where cells can be cultivated and exposed to multiple inputs; and 3) obtain temporal gene expression profiles using microarrays that have been exposed to combinatorial and temporally varying mixtures of stress mediators that closely mimic the physiological stress response. We hope to eventually use this information to predict the molecular events that determine a cell's progression to recovery or failure during stress.

Reception follows.