High-content assays for microbial and human cellulomics to assess sub-lethal pesticide effects.

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Our group has been studying the relationship between microbial function and cell surface properties. The cell surface is the first site of exposure of any cell to its external environment. With ever increasing environmental contaminants, such as pesticides, assessing their effect on human health and the microbial environment becomes paramount. We propose that sub-lethal exposure to pesticides can directly alter cell biochemical, physical properties and surface ultrastructure, ultimately affecting cellular immunity and microbial virulence and playing a greater role in environmental dysfunction and human disease than is currently appreciated. We will continue to develop models for the cell walls of select microorganisms and human cell membranes by characterizing their surface biochemical, ultrastructural and physical properties. To further understand the interplay between surface integrity and cell function, we will develop in vitro high-content assays. Single live cell analyses will be developed using atomic force microscopy, force spectroscopy and confocal microscopy to simultaneously monitor cell phenotype, ultrastructural morphology, cell mechanical properties and biochemistry, respectively. Metabolomics will be used to evaluate molecular changes to cells exposed to pesticides to identify "early warning" biomarkers indicative of progression towards cellular dysfunction. High content assays will be used to probe microbes, their cell wall biosynthetic mutants, and human cells in response to sub-lethal pesticide levels. Taken together, this multi-parameter biochemical, structural and physical data will be used to evaluate cellular responses to pesticides. This program will enhance our understanding of cell wall and membrane function, and their role in first line responses to external stimuli ultimately capable of altering cellular function. Once we fully characterize pesticide effects on model microbes and human cells, future studies can address their effects on pathogen-host interactions. Understanding the impact of environmental contaminants on the broader microbial ecosystem and the health risk of these agents to humans will have ultimate application in environmental protection and disease prevention.