title: Elucidating mechanisms of toxicity using phenotypic data from primary human cell systems--a chemical biology approach for thrombosis-related side effects.
abstract: Here we describe a chemical biology approach for elucidating potential toxicity mechanisms for thrombosis-related side effects. This work takes advantage of a large chemical biology data set comprising the effects of known, well-characterized reference agents on the cell surface levels of tissue factor (TF) in a primary human endothelial cell-based model of vascular inflammation, the BioMAP® 3C system. In previous work with the Environmental Protection Agency (EPA) for the ToxCast™ program, aryl hydrocarbon receptor (AhR) agonists and estrogen receptor (ER) antagonists were found to share an usual activity, that of increasing TF levels in this system. Since human exposure to compounds in both chemical classes is associated with increased incidence of thrombosis-related side effects, we expanded this analysis with a large number of well-characterized reference compounds in order to better understand the underlying mechanisms. As a result, mechanisms for increasing (AhR, histamine H1 receptor, histone deacetylase or HDAC, hsp90, nuclear factor kappa B or NFκB, MEK, oncostatin M receptor, Jak kinase, and p38 MAPK) and decreasing (vacuolar ATPase or V-ATPase) and mTOR) TF expression levels were uncovered. These data identify the nutrient, lipid, bacterial, and hypoxia sensing functions of autophagy as potential key regulatory points controlling cell surface TF levels in endothelial cells and support the mechanistic hypothesis that these functions are associated with thrombosis-related side effects in vivo.
authors.affiliation: BioSeek, a Division of DiscoveRx Corp., 310 Utah Ave., Suite 100, South San Francisco, CA 94080, USA. eberg@bioseekinc.com.
authors.forename: Ellen L
authors.initials: EL
authors.lastname: Berg
authors.affiliation: BioSeek, a Division of DiscoveRx Corp., 310 Utah Ave., Suite 100, South San Francisco, CA 94080, USA. mpolokoff@bioseekinc.com.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Polokoff
authors.affiliation: BioSeek, a Division of DiscoveRx Corp., 310 Utah Ave., Suite 100, South San Francisco, CA 94080, USA. aomahony@bioseekinc.com.
authors.forename: Alison
authors.initials: A
authors.lastname: O'Mahony
authors.affiliation: BioSeek, a Division of DiscoveRx Corp., 310 Utah Ave., Suite 100, South San Francisco, CA 94080, USA. dnguyen@bioseekinc.com.
authors.forename: Dat
authors.initials: D
authors.lastname: Nguyen
authors.affiliation: BioSeek, a Division of DiscoveRx Corp., 310 Utah Ave., Suite 100, South San Francisco, CA 94080, USA. xli@bioseekinc.com.
authors.forename: Xitong
authors.initials: X
authors.lastname: Li
mesh_terms: D001343:Autophagy; D015415:Biomarkers / Q000378:metabolism; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D008954:Models, Biological; D009930:Organic Chemicals / Q000633:toxicity*; D018336:Receptors, Aryl Hydrocarbon / Q000819:agonists / Q000378:metabolism; D011960:Receptors, Estrogen / Q000037:antagonists & inhibitors / Q000378:metabolism; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D013925:Thromboplastin / Q000378:metabolism; D013927:Thrombosis / Q000209:etiology*
chemical_list: D015415:Biomarkers; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D009930:Organic Chemicals; D018336:Receptors, Aryl Hydrocarbon; D011960:Receptors, Estrogen; D013925:Thromboplastin; D058570:TOR Serine-Threonine Kinases