You are here:
Adverse Outcome Pathways (AOPs) in Human Systems Biology: Gas-Phase Probes for Assessing In Vitro Enzyme System Perturbations
Citation:
Pleil, J., M. Angrish, AND M. Madden. Adverse Outcome Pathways (AOPs) in Human Systems Biology: Gas-Phase Probes for Assessing In Vitro Enzyme System Perturbations. PITTCON Confererence & Expo 2015, New Orleans, LA, March 08 - 12, 2015.
Impact/Purpose:
The National Exposure Research Laboratory (NERL) Human Exposure and Atmospheric Sciences Division (HEASD) conducts research in support of EPA mission to protect human health and the environment. HEASD research program supports Goal 1 (Clean Air) and Goal 4 (Healthy People) of EPA strategic plan. More specifically, our division conducts research to characterize the movement of pollutants from the source to contact with humans. Our multidisciplinary research program produces Methods, Measurements, and Models to identify relationships between and characterize processes that link source emissions, environmental concentrations, human exposures, and target-tissue dose. The impact of these tools is improved regulatory programs and policies for EPA.
Description:
The Air, Climate, and Energy (ACE) and Chemical Safety for Sustainability (CSS) programs at the U.S. EnvironmentalProtection Agency (EPA) encompass broad-based research that includes assessment of the health and environmentalimpacts of anthropogenic and manufactured chemicals. One component of these programs is to develop efficient andaccurate analytical methods for testing the relative toxicity and potential health effects of chemicals (and chemicalmixtures) and to ultimately guide chemical prioritization for more detailed evaluation and regulation. Currently, highthroughputin vitro methods using specific targeted adverse outcome pathways (AOPs) are implemented at the cellularand molecular level with liquid phase micro-fluidics including liquid chromatography-tandem mass spectrometry (LCMS/MS)analyses. New methodology is now under development to implement gas phase probe molecules for assessingspecific metabolic pathways that have already been well defined in human or mammalian models and apply semi-realtime detectors (optical or MS) to in vitro measurements. We propose two probe molecule and metabolite pairs thatcould be used for in vitro assessment of perturbations in specific liver enzyme systems: methyl-tertiary butyl ether(MTBE) metabolism to tertiary butyl alcohol (TBA) via CYP2A6 and sevoflurane metabolism to hexafluoroisopropanol viaCYP2E1 in flow through-reactors using human cell lines. We expect to extend the gas-phase probe methodology to airwayepithelial cells and analyze carbon monoxide (CO) production via heme oxygenase (HO) activity.
