Computational Model of Steroidogenesis in Human H295R Cells toPredict Biochemical Response to Endocrine-Active Chemicals: Model Developmentfor Metyrapone

摘要

Background: An in vitro steroidogenesis assay using the human adrenocortical carcinoma cell line H295R is being evaluated as a possible screening assay to detect and assess the impact of endocrine-active chemicals (EACs) capable of altering steroid biosynthesis. Data interpretation and their quantitative use in human and ecological risk assessments can be enhanced with mechanistic computational models to help define mechanisms of action and improve understanding of intracellular concentrationresponse behavior.Objectives: The goal of this study was to develop a mechanistic computational model of the metabolic network of adrenal steroidogenesis to estimate the synthesis and secretion of adrenal steroids in human H295R cells and their biochemical response to steroidogenesis-disrupting EAC.Methods: We developed a deterministic model that describes the biosynthetic pathways for the conversion of cholesterol to adrenal steroids and the kinetics for enzyme inhibition by metryrapone (MET), a model EAC. Using a nonlinear parameter estimation method, the model was fitted to the measurements from an in vitro steroidogenesis assay using H295R cells.Results: Model-predicted steroid concentrations in cells and culture medium corresponded well to the time-course measurements from control and MET-exposed cells. A sensitivity analysis indicated the parameter uncertainties and identified transport and metabolic processes that most influenced theconcentrations of primary adrenal steroids, aldosterone and cortisol.Conclusions: Our study demonstrates the feasibility of using a computationalmodel of steroidogenesis to estimate steroid concentrations invitro. This capability could be useful to help define mechanisms ofaction for poorly characterized chemicals and mixtures in support of predictivehazard and risk assessments with EACs.