The Core D proteomics core facility will support the proposed project to address the central hypothesis of the program that Protein glycosylation and glycoprotein remodeling modulate the coagulopathy and inflammation of sepsis. Core D will provide blood proteomic analyses in unbiased total and directed glycoproteomic approaches to identify various and specific changes to blood plasma proteomes. Plasma proteomic analyses will be first obtained and compared among uninfected plasma samples of wild-type mice and littermates bearing specific genetic deficiency states as indicated in the projects. Core D will contribute to achieving relevant project research aims in part by identifying and quantifying plasma proteins that are remodeled and regulated in abundance and activity by a newly discovered mechanism of secreted glycoprotein aging and turnover. These studies involve mice receiving glycosidase inhibitors, lacking glycosidases, and lacking endocytic lectin receptors that regulate blood plasma glycoprotein homeostasis. Core D will further provide plasma proteomic analyses in mouse sepsis arising from infections with different bacterial pathogens including Streptococcus pneumoniae (SPN), Salmonella enterica Typhimurium (ST), Salmonella enterica Choleraesuis (SC) and Escherichia coli (EC). Changes to the proteome and to specific features of the glycoproteome will be analyzed in these sepsis models and compared with proteomic and glycoproteomic changes that may occur in a model of the Systemic Inflammatory Response Syndrome (SIRS). Total plasma proteomic studies will also be performed among healthy human volunteers as well as those diagnosed with SIRS or sepsis caused by Gram-negative (mostly EC) or Gram-positive (including pneumococcal) bacterial pathogens. Core D will also contribute to achieving project research aims by identifying heparan sulfate proteoglycans (HSPGs) that are shed into the bloodstream in sepsis, and will further identify and quantify proteins that are bound to the glycosaminoglycans of circulating and shed HSPGs. Core D will further use activity-based proteomics to support project research to identify matrix metalloproteinases participating in the coagulopathy, inflammation, and outcomes of sepsis and SIRS. Together and in the aforementioned assays, Core D will act in part as a discovery engine to identify circulating blood glycoprotein targets that contribute to mechanisms modulating the coagulopathy and inflammation of sepsis. Altogether Core D will promote the attainment of synergistic research findings that integrate data from all three projects in addressing the central hypothesis of the program to further understand the onset and progression of the coagulopathy and inflammation of sepsis.
This core facility supports multiple research projects investigating recent discoveries pertaining to the pathogenesis of sepsis, which is a highly lethal syndrome afflicting millions of people in the U.S. each year and costing many billions of dollars in health care. Support by this core facility will enable project research goals focused on the debilitating and often lethal complications of sepsis that arise from host inflammation and coagulopathy.
