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Characterisation and validation of an in vitro transactivation assay based on the 22Rv1/MMTV_GR-KO cell line to detect human androgen receptor agonists and antagonists. | LitMetric

AI Article Synopsis

  • The study focuses on the development and validation of an androgen receptor (AR) transactivation assay, utilizing a specific prostate cancer cell line to detect AR agonists and antagonists.
  • Validation was conducted according to OECD guidelines, confirming that the assay performed comparably to an existing test, and showed no interference from glucocorticoid receptors.
  • The assay exhibited 100% concordance in chemical classifications across multiple labs, with strong accuracy and reliability metrics, leading to its approval for inclusion in the OECD testing guidelines in 2020.

Article Abstract

We describe the characterisation and validation of an androgen receptor (AR) transactivation assay for detection of AR agonists and antagonists using a stably transfected human prostate cancer cell line. This 22Rv1/mouse mammary tumour virus glucocorticoid knock-out cell line based AR transactivation assay was validated by criteria in Organisation for Economic Cooperation and Development Guidance Document 34 to determine if the assay performed equally well to the AR EcoScreen Assay included in Test Guideline for AR Transactivation (OECD TG 458). There was no Glucocorticoid Receptor (GR) crosstalk, and no changes in the AR DNA sequence in cells after the successful knock out of GR. Subsequently, the concordance of classifications of the 22 test chemicals was 100% in all laboratories. The AR agonistic and antagonistic inter-laboratory coefficients of variation based on log[10% effect for 10 nM DHT, PC] and log[inhibitory response of 800 pM DHT by at 30%, IC] from comprehensive tests were 2.75% and 2.44%, respectively. The AR agonist/antagonist test chemical classifications were consistent across AR EcoScreen ARTA assay data for 82/89%, and the balanced accuracy, sensitivity, and specificity were 83/90%, 88/100% and 78/80%, respectively. This assay was successfully validated and was approved for inclusion in TG 458 in 2020.

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Source
http://dx.doi.org/10.1016/j.fct.2021.112206DOI Listing

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