title: Evaluation of perfluoroalkyl acid activity using primary mouse and human hepatocytes.
abstract: While perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) have been studied at length, less is known about the biological activity of other perfluoroalkyl acids (PFAAs) detected in the environment. Using a transient transfection assay developed in COS-1 cells, our group has previously evaluated a variety of PFAAs for activity associated with activation of peroxisome proliferator-activated receptor alpha (PPARα). Here we use primary heptatocytes to further assess the biological activity of a similar group of PFAAs using custom designed Taqman Low Density Arrays. Primary mouse and human hepatoyctes were cultured for 48h in the presence of varying concentrations of 12 different PFAAs or Wy14,643, a known activator of PPARα. Total RNA was collected and the expression of 48 mouse or human genes evaluated. Gene selection was based on either in-house liver microarray data (mouse) or published data using primary hepatocytes (human). Gene expression in primary mouse hepatocytes was more restricted than expected. Genes typically regulated in whole tissue by PPARα agonists were not altered in mouse cells including Acox1, Me1, Acaa1a, Hmgcs1, and Slc27a1. Cyp2b10, a gene regulated by the constitutive androstane receptor and a transcript normally up-regulated by in vivo exposure to PFAAs, was also unchanged in cultured mouse hepatocytes. Cyp4a14, Ehhadh, Pdk4, Cpt1b, and Fabp1 were regulated as expected in mouse cells. A larger group of genes were differentially expressed in human primary hepatocytes, however, little consistency was observed across compounds with respect to which genes produced a significant dose response making the determination of relative biological activity difficult. This likely reflects weaker activation of PPARα in human versus rodent cells as well as variation among individual cell donors. Unlike mouse cells, CYP2B6 was up-regulated in human hepatocytes by a number of PFAAs as was PPARδ. Rankings were conducted on the limited dataset. In mouse hepatocytes, the pattern was similar to that previously observed in the COS-1 reporter cell assay. With the exception of PFHxA, longer chain PFAA carboxylates were the most active. The pattern was similar in human hepatocytes, although PFDA and PFOS showed higher activity than previously observed while PFOA showed somewhat less activity. These data reflect inherent challenges in using primary hepatocytes to predict toxicological response.
authors.affiliation: U.S. Environmental Protection Agency, Office of Research and Development, National Health and Environmental Effects Research Laboratory, Integrated Systems Toxicology Division, Research Triangle Park, NC 27711, USA. rosen.mitch@epa.gov
authors.forename: Mitchell B
authors.initials: MB
authors.lastname: Rosen
authors.forename: Kaberi P
authors.initials: KP
authors.lastname: Das
authors.forename: Carmen R
authors.initials: CR
authors.lastname: Wood
authors.forename: Cynthia J
authors.initials: CJ
authors.lastname: Wolf
authors.forename: Barbara D
authors.initials: BD
authors.lastname: Abbott
authors.forename: Christopher
authors.initials: C
authors.lastname: Lau
mesh_terms: D017738:Alkanesulfonic Acids / Q000737:chemistry* / Q000633:toxicity*; D000818:Animals; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005466:Fluorocarbons / Q000737:chemistry* / Q000633:toxicity*; D022781:Hepatocytes / Q000187:drug effects*; D006801:Humans; D051379:Mice; D061251:Primary Cell Culture
chemical_list: D017738:Alkanesulfonic Acids; D005466:Fluorocarbons; C076994:perfluorooctane sulfonic acid