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Proteomics for Adverse Outcome Pathway Discovery using Human Kidney Cells?
Citation:
VanEmon, J., F. van Breukelen, AND P. Pan. Proteomics for Adverse Outcome Pathway Discovery using Human Kidney Cells? Pittcon 2016, Atlanta, GA, March 06 - 10, 2016.
Impact/Purpose:
Presented at Pittcon 2016 in Atlanta, GA.
Description:
An Adverse Outcome Pathway (AOP) is a conceptual framework that applies molecular-based data for use in risk assessment and regulatory decision support. AOP development is based on effects data of chemicals on biological processes (i.e., molecular initiating events, key intermediate toxicity events, proteomic changes) to link exposure to effects. Cell culture exposure research has proven to be an effective route for aiding the development of specific AOPs. Here we describe a proteomic analysis to study the effects of chlorpyrifos and 3,5,6-trichloro-2-pyridinol (TCP) on human kidney cells. HEK293 (CRL-1573) ATTC cells were initially cultured in DMEM. Cells were next seeded at 0.5 million cells per well in 6-well plates and cultured overnight. The following day cells were exposed to various levels of chlorpyrifos and TCP for 24 hours. Protein expression patterns were obtained before and after dosing. Over 100 peptides were identified using high-resolution mass spectrometry and assignments to 372 proteins were made. Groups of proteins could consistently be assigned as being either up-regulated or down-regulated in response to the dosing levels of the compounds. The grouping of proteins changed based on exposure to either single analyte or tandem analyte exposure. The proteomic analysis correlated well with cytotoxicity data that was also obtained leading to potential biomarkers of exposure identification.
