Living Cell Arrays for Real Time Functional Genomics

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DESCRIPTION (provided by applicant): New tools are revolutionizing 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 on a large scale is notable. However, it is often difficult to correlate the trends and relationships observed in normal and abnormal states to the phenotype resulting from the gene expression profile. For physiological states involving metabolic derangements, gene expression profiling does not fully explain the complex molecular mechanisms involved. Thus, in order to develop a comprehensive understanding of metabolic states, it is essential to understand both the gene expression events as well as the cytoplasmic events, which control changes in metabolites. The proposed Bioengineering Research Partnership seeks to develop a new functional genomics approach for studying gene expression: the use of intact cells for the simultaneous temporal expression profiling of multiple genes using aequorin-type fluorescent proteins (AFP) in a massively parallel, high throughput format. The specific aims are: (1) to generate and characterize a panel of reporter cell lines that monitors the major events in the inflammatory cytokine signaling cascades; (2) to design a microfluidic system to dynamically control the input stimulus as well as the fluorescent response of an array of primary rat hepatocytes and H35 hepatoma reporter cell lines; (3) to characterize the dynamics of cytokine signal transduction and the impact of steatosis and different metabolic states on the acute phase response of these cells. This project will be carried out by three distinct research groups, which will interact extensively. Dr. Martin Yarmush (MGH), the Principal Investigator, will oversee the Administrative and Technical Core, and lead the Cell Physiology and Imaging group, which will integrate the technologies developed by the other Pl's and carry out the bulk of the cell physiology experiments. Dr. Mehmet Toner, Director of the Microscale Engineering Facility at the MGH, will lead the Microfabrication and Microfluidics group focusing on the development of microfabricated devices. Dr. Jeffrey Morgan (Brown Univ., Providence, RI), a molecular biologist and expert in gene therapy, will lead the Molecular Biology and Cell Analysis group focusing on reporter cell line development.