The Cellarium is a new class of microarray-based technology that produces dynamic, high-throughput metabolic measurements of live single cells. Years 1 and 2 of the U01 Cellarium program have been marked by significant progress toward development of this microarray approach which will enable generation of unique data types (single cell physiology over time) for integration into the Library of Integrated Network-Based Cellular Signatures (LINCS). We propose to use the Cellarium to investigate the biological relevance of cellular heterogeneity in cancer- and inflammation-related metabolic regulation. This will demonstrate that the technology can be made applicable to a broad variety of biological questions. In collaboration with Columbia University we have begun to explore the effects of predicted master regulator genes of the Warburg effect. We will continue this effort in Yr3 and will select at least one newly identified regulatory gene and to fully analyze its metabolic manifestation at the single cell level in the Cellarium. In addition, we will extend the parent technology by adding intracellular K+ and extracellular tandem K+/O2 sensing capability to our current suite of sensors (O2, pH, glucose and ATP). We will validate these new capabilities and use them to study the role of cellular heterogeneity in inflammatory response. To this end, we will analyze both O2 consumption and K+ fluxes in nave and activated macrophages and correlate the results with inflammasome/pyroptosome formation. This Cellarium technology enhancement will improve translational potential which we will demonstrate by application to biomedically relevant questions. We will continue to facilitate Cellarium data availability and incorporation into LINCS by engaging and working closely with the Computational U01 LINCS Centers.
|Effective start/end date||9/16/11 → 8/31/14|
- HHS-NIH: National Cancer Institute (NCI): $1,509,670.00