Activity study of possible endocrine disruption via the TRb or PPARy2 receptors by using CALUX cell systems

Commonly used acronym: TRb and PPARy2 CALUX

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Scope of the method

The Method relates to
  • Human health
The Method is situated in
  • Basic Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Human derived cells / tissues / organs
Specify the type of cells/tissues/organs
U2-OS cells (human osteoblasts), stably transfected with the receptor and a luciferase reporter construct

Description

Method keywords
  • cell culture
  • CALUX
  • reporter gene assays
Scientific area keywords
  • Endocrine disruptors
  • obesogens
  • PPARy
  • TRb
  • mixtures
  • concentration addition
Method description

The CALUX® system (Chemically Actived LUciferase eXpression) of Bio Detection Systems (BDS, Amsterdam, the Netherlands) uses U-2 OS cells (human osteoblast) that are stably transfected with human TRb or human PPARg2 (BDS, Amsterdam) and a luciferase reporter construct under the control of a receptor specific response element. Through measuring the activity of chemical compounds on these cell lines, we can determine if they can potentially have endocrine activity. The activity of mixture of chemicals can also be determined in these cell systems.

Lab equipment
  • Cell incubator,
  • Safety cabinet,
  • Fluorimeter.
Method status
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages
  • Speed (vs in vivo experiments),
  • Useful for prioritisation.
Challenges

Difficult to extrapolate to the in vivo situation because of Absorption, Distribution, Metabolism, and Excretion (ADME).

References, associated documents and other information

References
  • Mertens, B, Van Heyst A, Demaegdt H, Boonen I, Van Den Houwe K, Goscinny S, Elskens M, and Van Hoeck E. Assessment of hazards and risks associated with dietary exposure to mineral oil for the Belgian population. Food and Chemical Toxicology, 2021, Volume 149
  • I Boonen I, Van Heyst A , Van Langenhove K,Van Hoeck E, Mertens M, Denison MS, Elskens M, Demaegdt H. Assessing the receptor-mediated activity of PAHs using AhR-, ERα- and PPARγ- CALUX bioassays. Food Chem Toxicol. 2020, 145:111602. doi: 10.1016/j.fct.2020.111602
  • Simon C, Onghena M, Covaci A, Van Hoeck E, Van Loco J, Vandermarken T, Van Langenhove K, Demaegdt H, Mertens B, Vandermeiren K, Scippo ML, Elskens M. Screening of endocrine activity of compounds migrating from plastic baby bottles using a multi-receptor panel of in vitro bioassays.
  • Mertens B, Van Hoeck E, Blaude MN, Simon C, Onghena M, Vandermarken T, Van Langenhove K, Demaegdt H, Vandermeiren K, Covaci A, Scippo ML, Elskens M, Van Loco J. Evaluation of the potential health risks of substances migrating from polycarbonate replacement baby bottles. Food Chem Toxicol. 2016, 97:108-119.
  • Demaegdt H, Daminet B, Evrard A, Scippo ML, Muller M, Pussemier L, Callebaut A, Vandermeiren K. Endocrine activity of mycotoxins and mycotoxin mixtures. Food Chem Toxicol. 2016, 96:107-16.
  • Pereira-Fernandes A, Demaegdt H, Vandermeiren K, Hectors TL, Jorens PG, Blust R, Vanparys C. Evaluation of a Screening System for Obesogenic Compounds: Screening of Endocrine Disrupting Compounds and Evaluation of the PPAR Dependency of the Effect. PLoS One. 2013, 8 : e77481.
Associated documents

Organisations

Sciensano
Chemical and physical health risks
Belgium

Partners

BDS