title: Pharmacotoxicology of clinically-relevant concentrations of obeticholic acid in an organotypic human hepatocyte system.
abstract: Nonalcoholic steatohepatitis (NASH) is an emerging health crisis with no approved therapies. Obeticholic acid (OCA), a farnesoid X receptor (FXR) agonist, shows promise in NASH trials. However, the precise mechanisms mediating OCA effects and impact on cholesterol metabolism are not fully understood. We explored the pharmaco-toxicological effects of OCA on patho-physiological pathways in hepatocytes using a previously described perfused organotypic liver system that allows culture in near-physiological insulin/glucose milieus, and exhibits drug responses at clinically-relevant concentrations. Primary hepatocytes experienced 48-hour exposure to OCA at concentrations approximating therapeutic (0.5μM) and supratherapeutic (10μM) levels. Global transcriptomics by RNAseq was complimented by cellular viability (MTT), CYP activity assays, and secreted FGF19 levels in the media. Dose-dependent, transcriptional effects suggested suppression of bile acid synthesis (↓CYP7A1, ↓CYP27A1) and increased bile efflux (↑ABCB4, ↑ABCB11, ↑OSTA, ↑OSTB). Pleiotropic effects included suppression of TGFβ and IL-6 signaling pathways, and signatures suggestive of HDL suppression (↑SCARB1, ↓ApoAI, ↓LCAT) and LDL elevation (↑ApoB, ↓CYP7A1). OCA exhibited direct FXR-mediated effects with increased FGF19 secretion. Transcriptomics revealed regulation of metabolic, anti-inflammatory, and anti-fibrotic pathways beneficial in NASH, and predicted cholesterol profiles consistent with clinical findings. Follow-up studies under lipotoxic/inflammatory conditions would corroborate these effects in a disease-relevant environment.
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States.
authors.forename: A
authors.initials: A
authors.lastname: Dash
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States.
authors.forename: R A
authors.initials: RA
authors.lastname: Figler
authors.affiliation: VL37, Inc., 790 Memorial Drive, Cambridge, MA, United States.
authors.forename: B R
authors.initials: BR
authors.lastname: Blackman
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States.
authors.forename: S
authors.initials: S
authors.lastname: Marukian
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States.
authors.forename: M S
authors.initials: MS
authors.lastname: Collado
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States.
authors.forename: M J
authors.initials: MJ
authors.lastname: Lawson
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States.
authors.forename: S A
authors.initials: SA
authors.lastname: Hoang
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States.
authors.forename: A J
authors.initials: AJ
authors.lastname: Mackey
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States.
authors.forename: D
authors.initials: D
authors.lastname: Manka
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States.
authors.forename: B K
authors.initials: BK
authors.lastname: Cole
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States.
authors.forename: R E
authors.initials: RE
authors.lastname: Feaver
authors.affiliation: Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, VA, United States.
authors.forename: A J
authors.initials: AJ
authors.lastname: Sanyal
authors.affiliation: HemoShear Therapeutics LLC, Charlottesville, VA, United States. Electronic address: Wamhoff@HemoShear.com.
authors.forename: B R
authors.initials: BR
authors.lastname: Wamhoff
mesh_terms: D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002635:Chenodeoxycholic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000633:toxicity; D002784:Cholesterol / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D065626:Non-alcoholic Fatty Liver Disease / Q000378:metabolism; D059467:Transcriptome / Q000187:drug effects
chemical_list: C464660:obeticholic acid; D002635:Chenodeoxycholic Acid; D002784:Cholesterol
keywords: FXR; Flow; Liver; NASH; Obeticholic; Transcriptomics