Discovery of phosgene and chlorine gas modes of action and therapeutic targets using chemoproteomic profiling strategies

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Project Summary: Phosgene and chlorine are two toxic gases produced on large industrial scales that pose occupational hazards and chemical threats to the public and military. Both toxicants modify proteins in various ways and induce oxidative stress in the lung which perturb critical functions and pathways. However, the precise nature and sites of these modifications in the lung proteome have not been identified, and the potential chemical diversity of these modifications poses an analytical challenge to unraveling the molecular mechanisms by which chlorine and phosgene induce acute lung injury. This lack of biomolecular characterization of phosgene- and chlorine-induced modification of protein targets has consequently hindered our ability to develop effective countermeasures; there are currently no approved therapeutics for treating phosgene or chlorine gas exposures. We propose to use chemoproteomic profiling—a subset of mass spectrometry-based proteomics that can detect the sites and types of distinct chemical modifications to proteins—to comprehensively identify specific proteins in the lung that have been chemically altered by phosgene and chlorine and perform pathway analysis to identify protein candidates for future targeted countermeasure development. The specific aims of this proposal are: Aim 1: Dynamic redox profiling to identify the temporal protein cysteine oxidation events and global proteome changes following gaseous toxicant exposure Aim 2: Mapping phosgene- and chlorine-induced protein modifications in the lung using electrophilic chemical probes Aim 3: Data integration and network analysis to identify shared and unique molecular targets of phosgene and chlorine for therapeutic development