@article{c7cc76b7669b4131914341569cbbea9c,
title = "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",
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, PC10] and log[inhibitory response of 800 pM DHT by at 30%, IC30] 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.",
keywords = "Agonist, Androgen receptor, Antagonist, OECD Test guideline, Transcriptional activation assay, Validation",
author = "Yooheon Park and Jung, {Da Woon} and Anne Milcamps and Masahiro Takeyoshi and Jacobs, {Miriam N.} and Houck, {Keith A.} and Atsushi Ono and Bovee, {Toine F.H.} and Patience Browne and Nathalie Delrue and Kang, {Yun Sook} and Lee, {Hee Seok}",
note = "Funding Information: The use of in vitro EAS assays can be used to selectively prioritise active chemicals for additional testing and therefore, supports the 3Rs principles to reduce animal testing for ethical and economic reasons and helps elucidate underlying mechanisms of action.This new ARTA assay based on the 22Rv1/MMTV_GR-KO cell line was proposed for validation as an internationally harmonised OECD test method to be included in OECD TG 458. Validation of this ARTA assay based on the 22Rv1/MMTV_GR-KO cell line, conducted in accordance with the OECD Guidance Document No. 34 (OECD, 2005), was managed by an independent Study Management Team (SMT), together with the lead laboratory, and with the support of the OECD expert group Validation Management Group for Non-Animal testing (VMG-NA). Here we present the results of the validation studies on this new ARTA assay to detect AR agonists/antagonists by three independent laboratories.As shown in Fig. 1, three independent laboratories, NIFDS, Korean Testing and Research Institute (KTR) and Dongguk University, participated in the full validation study. For the full validation study, data for test chemical was obtained from three valid pre-screen runs, followed by three valid comprehensive runs. All validation processes were conducted according to Good Laboratory Practices (GLP) principles. SMT members of the OECD VMG-NA provided independent management that included oversight and guidance, and a member of the OECD VMG-NA provided additional support characterising potential AR-GR crosstalk and the cell-line construct.This research was supported in Republic of Korea by grants 18161MFDS108 from the Ministry of Food and Drug Safety. Funding Information: This research was supported in Republic of Korea by grants 18161MFDS108 from the Ministry of Food and Drug Safety . Publisher Copyright: {\textcopyright} 2021 Elsevier Ltd",
year = "2021",
month = jun,
doi = "10.1016/j.fct.2021.112206",
language = "English",
volume = "152",
journal = "Food and Chemical Toxicology",
issn = "0278-6915",
publisher = "Elsevier Limited",
}
