Biology Working Groups

Three biology working groups (BWG) will address key questions of contemporary heart, blood and lung research and provide the interface between the technology development and applied activities of the Center. The groups bring together distinguished researchers from the Puget Sound area with the purpose of identifying novel approaches to address biological problems utilizing proteomics. The three working groups will focus on:

Macrophage activation
This group recognizes the critical protective role played by monocyte/macrophage as a primary means of defense in an inflammatory response through its capacity as the scavenger cell of the body and its fundamental importance in specific immunity. However, inflammation also forms the basis of a series of life-threatening and functionally crippling diseases that includes atherosclerosis and fibrotic lung diseases. In particular, the activated macrophage can release an incredible array of proteins that can induce tissue damage, promote the influx of more leukocytes and cause extensive scarring and fibrosis that can result in occlusion of blood vessels and lung alveoli. Although a number of checks and balances prevent this destructive potential from being realized in normal responses to injury, they are insufficient to block the progressive destruction in chronic inflammatory diseases. Thus, defining the comprehensive proteome of an activated macrophage is of primary importance to identify strategies to regulate the destructive pathways without significantly impairing protective mechanisms.

Cardiovascular and blood cell development
This group is based on the insight that different cells in an organism are defined by the complement of genes that they express. Cell fate choices reflect and require the initiation of new programs of gene expression (and extinguishing of others) that once initiated, must be faithfully passed from parental cells to their progeny. Therefore, to appreciate the complex processes of mammalian development and potentially apply this knowledge to gene therapy for genetic disorders, it is essential to understand how a cell determines which genes it will express. A major unresolved question is how these changes are initiated and propagated at specific gene loci during development. The goal of this area of investigation is to use mass spectrometry to interrogate these changes during the differentiation of specific cardiac and hematopoietic cell lineages. The development of technological approaches and general models established from these studies should provide a basis for studying tissue-specific gene expression at multiple loci, including those of the macrophage that is the long-term focus of this program.

Signaling
The Signaling Working Group recognizes that changes in signaling pathways and the composition of signaling complexes are critical to the initiation of contraction, proliferation, activation, and apoptosis - processes involved in a variety of cardiovascular diseases. Many of the same signaling pathways have been implicated in all of these processes, and yet the stimuli and downstream effectors are distinct. The proposed studies will focus on the signaling pathways in cardiac myocytes, vascular smooth muscle, fibroblasts and endothelial cells whose signaling pathways share many common themes. Hypothesis-driven analysis of signaling complexes and more discovery-based approaches such as yeast two-hybrid analysis have demonstrated that critical players in particular signaling complexes interact directly with diverse groups of proteins, including protein kinases, kinase anchoring proteins, scaffolding proteins and cell adhesion molecules. However, definition of these complexes is far from complete. Particular signaling pathways, such as those associated with membranes, have been especially difficult to approach experimentally. The goal of this working group is to try to expand our ability to probe signaling pathways and complexes by using mass spectrometry to: 1) more completely define the changes in cellular protein profiles associated with major physiologic responses; 2) detail the components of specific signaling complexes and determine how they dynamically change with different physiologic stimuli; 3) determine the state of phosphorylation of signaling complexes and changes in the protein phosphorylation profile of proteins in response to physiologic stimuli; and 4) develop new approaches for examining protein interactions in signaling pathways highly regulated by their membrane interactions.

It should be noted that the projects of the BWGs are not directly funded by the Center. The Center will, however, make capacity for sample processing available via the HTPF.