{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T13:14:34Z","timestamp":1787750074233,"version":"build-2784847793"},"reference-count":70,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,2,28]],"date-time":"2024-02-28T00:00:00Z","timestamp":1709078400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Toxicol."],"abstract":"<jats:p><jats:bold>Introduction:<\/jats:bold> Skin sensitization, which leads to allergic contact dermatitis, is a key toxicological endpoint with high occupational and consumer prevalence. This study optimized several <jats:italic>in vitro<\/jats:italic> assays listed in OECD skin sensitization test guidelines for use on a quantitative high-throughput screening (qHTS) platform and performed <jats:italic>in silico<\/jats:italic> model predictions to assess the skin sensitization potential of prioritized compounds from the Tox21 10K compound library.<\/jats:p><jats:p><jats:bold>Methods:<\/jats:bold> First, we screened the entire Tox21 10K compound library using a qHTS KeratinoSens<jats:sup>TM<\/jats:sup> (KS) assay and built a quantitative structure\u2013activity relationship (QSAR) model based on the KS results. From the qHTS KS screening results, we prioritized 288 compounds to cover a wide range of structural chemotypes and tested them in the solid phase extraction\u2013tandem mass spectrometry (SPE\u2013MS\/MS) direct peptide reactivity assay (DPRA), IL-8 homogeneous time-resolved fluorescence (HTRF) assay, CD86 and CD54 surface expression in THP1 cells, and predicted <jats:italic>in silico<\/jats:italic> sensitization potential using the OECD QSAR Toolbox (v4.5).<\/jats:p><jats:p><jats:bold>Results:<\/jats:bold> Interpreting tiered qHTS datasets using a defined approach showed the effectiveness and efficiency of <jats:italic>in vitro<\/jats:italic> methods. We selected structural chemotypes to present this diverse chemical collection and to explore previously unidentified structural contributions to sensitization potential.<\/jats:p><jats:p><jats:bold>Discussion:<\/jats:bold> Here, we provide a skin sensitization dataset of unprecedented size, along with associated tools, and analysis designed to support chemical assessments.<\/jats:p>","DOI":"10.3389\/ftox.2024.1321857","type":"journal-article","created":{"date-parts":[[2024,2,28]],"date-time":"2024-02-28T05:19:00Z","timestamp":1709097540000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Use of in vitro methods combined with in silico analysis to identify potential skin sensitizers in the Tox21 10K compound library"],"prefix":"10.3389","volume":"6","author":[{"given":"Zhengxi","family":"Wei","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tuan","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Judy","family":"Strickland","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuhong","family":"Fang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dingyin","family":"Tao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anton","family":"Simeonov","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruili","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicole C.","family":"Kleinstreuer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Menghang","family":"Xia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,2,28]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"223","DOI":"10.3109\/1547691X.2012.711782","article-title":"Danger, intracellular signaling, and the orchestration of dendritic cell function in skin sensitization","volume":"10","author":"Ainscough","year":"2013","journal-title":"J. Immunotoxicol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"262","DOI":"10.1016\/j.taap.2014.12.014","article-title":"Predicting chemically-induced skin reactions. Part I: QSAR models of skin sensitization and their application to identify potentially hazardous compounds","volume":"284","author":"Alves","year":"2015","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"B3","doi-asserted-by":"publisher","first-page":"733","DOI":"10.1016\/j.tiv.2010.12.014","article-title":"The intra-and inter-laboratory reproducibility and predictivity of the KeratinoSens assay to predict skin sensitizers in vitro: results of a ring-study in five laboratories","volume":"25","author":"Andreas","year":"2011","journal-title":"Toxicol. Vitro"},{"key":"B4","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1097\/DER.0000000000000003","article-title":"Categorization of chemicals according to their relative human skin sensitizing potency","volume":"25","author":"Basketter","year":"2014","journal-title":"Dermatitis"},{"key":"B5","doi-asserted-by":"publisher","first-page":"054501","DOI":"10.1289\/EHP1759","article-title":"An integrated chemical environment to support 21st-century toxicology","volume":"125","author":"Bell","year":"2017","journal-title":"Environ. health Perspect."},{"key":"B6","doi-asserted-by":"publisher","first-page":"e54750","DOI":"10.3791\/54750","article-title":"An efficient method for the synthesis of peptoids with mixed lysine-type\/arginine-type monomers and evaluation of their anti-leishmanial activity","author":"Bolt","year":"2016","journal-title":"JoVE J. Vis. Exp."},{"key":"B7","doi-asserted-by":"publisher","first-page":"1013","DOI":"10.1021\/acs.jcim.7b00194","article-title":"Pred-skin: a fast and reliable web application to assess skin sensitization effect of chemicals","volume":"57","author":"Braga","year":"2017","journal-title":"J. Chem. Inf. Model."},{"key":"B8","doi-asserted-by":"publisher","first-page":"1403","DOI":"10.1007\/s00216-021-03766-x","article-title":"Suspect and non-target screening of chemicals in clothing textiles by reversed-phase liquid chromatography\/hybrid quadrupole-Orbitrap mass spectrometry","volume":"414","author":"Carlsson","year":"2022","journal-title":"Anal. Bioanal. Chem."},{"key":"B9","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1007\/s00204-017-2097-4","article-title":"Standardisation of defined approaches for skin sensitisation testing to support regulatory use and international adoption: position of the International Cooperation on Alternative Test Methods","volume":"92","author":"Casati","year":"2018","journal-title":"Archives Toxicol."},{"key":"B10","first-page":"1","article-title":"Xgboost: extreme gradient boosting","author":"Chen","year":"2015","journal-title":"R. package version 0"},{"key":"B11","doi-asserted-by":"publisher","first-page":"17654","DOI":"10.1074\/jbc.RA119.010201","article-title":"Physiologically relevant orthogonal assays for the discovery of small-molecule modulators of WIP1 phosphatase in high-throughput screens","volume":"294","author":"Clausse","year":"2019","journal-title":"J. Biol. Chem."},{"key":"B12","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1016\/j.yrtph.2018.03.003","article-title":"International regulatory requirements for skin sensitization testing","volume":"95","author":"Daniel","year":"2018","journal-title":"Regul. Toxicol. Pharmacol."},{"key":"B13","first-page":"1.5","volume-title":"Misc Functions of the Department of Statistics (e1071","author":"Dimitriadou","year":"2005"},{"key":"B14","volume-title":"Interim science policy: use of alternative approaches for skin sensitization as a replacement for laboratory animal testing draft for public comment","year":"2018"},{"key":"B15","doi-asserted-by":"publisher","first-page":"164","DOI":"10.1093\/toxsci\/kfp192","article-title":"Investigation of peptide reactivity of pro-hapten skin sensitizers using a peroxidase-peroxide oxidation system","volume":"112","author":"Gerberick","year":"2009","journal-title":"Toxicol. Sci."},{"key":"B16","doi-asserted-by":"publisher","first-page":"332","DOI":"10.1093\/toxsci\/kfh213","article-title":"Development of a peptide reactivity assay for screening contact allergens","volume":"81","author":"Gerberick","year":"2004","journal-title":"Toxicol. Sci."},{"key":"B17","doi-asserted-by":"publisher","first-page":"33","DOI":"10.14573\/altex.1911261","article-title":"Evaluation of the global performance of eight in silico skin sensitization models using human data","volume":"38","author":"Golden","year":"2020","journal-title":"Altex"},{"key":"B18","doi-asserted-by":"publisher","first-page":"104985","DOI":"10.1016\/j.envint.2019.104985","article-title":"Exposure of metals and PAH through local foods and risk of cancer in a historically contaminated glassworks area","volume":"131","author":"Helmfrid","year":"2019","journal-title":"Environ. Int."},{"key":"B19","doi-asserted-by":"publisher","first-page":"1333","DOI":"10.1002\/jat.3105","article-title":"Evaluation of combinations of in vitro sensitization test descriptors for the artificial neural network-based risk assessment model of skin sensitization","volume":"35","author":"Hirota","year":"2015","journal-title":"J. Appl. Toxicol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"344","DOI":"10.1080\/10408444.2018.1429385","article-title":"Non-animal methods to predict skin sensitization (I): the Cosmetics Europe database<sup\/>","volume":"48","author":"Hoffmann","year":"2018","journal-title":"Crit. Rev. Toxicol."},{"key":"B21","doi-asserted-by":"crossref","DOI":"10.1007\/978-1-4939-6346-1_12","article-title":"A quantitative high-throughput screening data analysis pipeline for activity profiling","volume-title":"High-throughput screening assays in Toxicology","author":"Huang","year":"2016"},{"key":"B22","volume-title":"ICCVAM test method evaluation report: usefulness and limitations of the murine local lymph node assay for potency categorization of chemicals causing allergic contact dermatitis in humans","year":"2011"},{"key":"B23","doi-asserted-by":"publisher","first-page":"11473","DOI":"10.1073\/pnas.0604348103","article-title":"Quantitative high-throughput screening: a titration-based approach that efficiently identifies biological activities in large chemical libraries","volume":"103","author":"Inglese","year":"2006","journal-title":"Proc. Natl. Acad. Sci."},{"key":"B24","doi-asserted-by":"publisher","first-page":"8","DOI":"10.2310\/6620.2009.09038","article-title":"Local lymph node data for the evaluation of skin sensitization alternatives: a second compilation","volume":"21","author":"Kern","year":"2010","journal-title":"DERM"},{"key":"B25","doi-asserted-by":"publisher","first-page":"S238","DOI":"10.1093\/toxsci\/kfq346","article-title":"Chemical allergy: translating biology into hazard characterization","volume":"120","author":"Kimber","year":"2011","journal-title":"Toxicol. Sci."},{"key":"B26","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1080\/10408444.2018.1429386","article-title":"Non-animal methods to predict skin sensitization (II): an assessment of defined approaches","volume":"48","author":"Kleinstreuer","year":"2018","journal-title":"Crit. Rev. Toxicol."},{"key":"B27","doi-asserted-by":"publisher","first-page":"375","DOI":"10.1038\/jid.1966.159","article-title":"The identification of contact allergens by human assay: II. Factors influencing the induction and measurement of allergic contact dermatitis","volume":"47","author":"Kligman","year":"1966","journal-title":"J. Investigative Dermatology"},{"key":"B28","first-page":"18","article-title":"Classification and regression by randomForest","volume":"2","author":"Liaw","year":"2002","journal-title":"R. news"},{"key":"B29","doi-asserted-by":"publisher","first-page":"885","DOI":"10.1007\/s10822-011-9472-7","article-title":"Fragment-based prediction of skin sensitization using recursive partitioning","volume":"25","author":"Lu","year":"2011","journal-title":"J. Comput. Aided Mol. Des."},{"key":"B30","doi-asserted-by":"publisher","first-page":"79","DOI":"10.32614\/rj-2014-008","article-title":"ROSE: a package for binary imbalanced learning","volume":"6","author":"Lunardon","year":"2014","journal-title":"R J."},{"key":"B31","doi-asserted-by":"publisher","first-page":"776","DOI":"10.1001\/jama.1946.02870260060020","article-title":"Toxicity of thioglycolic acid used in cold permanent wave process","volume":"131","author":"McCord","year":"1946","journal-title":"J. Am. Med. Assoc."},{"key":"B32","doi-asserted-by":"publisher","first-page":"1470","DOI":"10.1016\/j.jaci.2006.12.654","article-title":"Chemokine responses distinguish chemical-induced allergic from irritant skin inflammation: memory T cells make the difference","volume":"119","author":"Meller","year":"2007","journal-title":"J. Allergy Clin. Immunol."},{"key":"B33","article-title":"para-Phenylenediamine testing in strongly sensitized patients: an effective screen: CD06","volume":"171","author":"Nalluri","year":"2014","journal-title":"Br. J. Dermatology"},{"key":"B34","doi-asserted-by":"publisher","first-page":"733","DOI":"10.1016\/j.tiv.2010.12.014","article-title":"The intra-and inter-laboratory reproducibility and predictivity of the KeratinoSens assay to predict skin sensitizers in vitro: results of a ring-study in five laboratories","volume":"25","author":"Natsch","year":"2011","journal-title":"Toxicol. Vitro"},{"key":"B35","doi-asserted-by":"publisher","first-page":"1150","DOI":"10.1016\/j.tiv.2012.07.001","article-title":"Prediction of skin sensitization potency of chemicals by human Cell Line Activation Test (h-CLAT) and an attempt at classifying skin sensitization potency","volume":"26","author":"Nukada","year":"2012","journal-title":"Toxicol. vitro"},{"key":"B36","doi-asserted-by":"publisher","first-page":"609","DOI":"10.1016\/j.tiv.2012.11.006","article-title":"Data integration of non-animal tests for the development of a test battery to predict the skin sensitizing potential and potency of chemicals","volume":"27","author":"Nukada","year":"2013","journal-title":"Toxicol. Vitro"},{"key":"B37","doi-asserted-by":"publisher","first-page":"175","DOI":"10.2131\/jts.33.175","article-title":"Production of IL-8 in THP-1 cells following contact allergen stimulation via mitogen-activated protein kinase activation or tumor necrosis factor-alpha production","volume":"33","author":"Nukada","year":"2008","journal-title":"J. Toxicol. Sci."},{"key":"B38","volume-title":"Test guideline 429: the local lymph node assay","year":"2002"},{"key":"B39","unstructured":""},{"key":"B40","volume-title":"Guidance document on the reporting of defined approaches to be used within integrated approaches to testing and assessment","year":""},{"key":"B41","volume-title":"Guidelines for the testing of chemicals section 4","year":"2019"},{"key":"B42","volume-title":"Guideline No. 497: defined approaches on skin sensitisation","year":"2021"},{"key":"B43","first-page":"442E","volume-title":"skin sensitisation:","year":"2023"},{"key":"B44","first-page":"442D","volume-title":"skin sensitisation: ARE-Nrf2 luciferase test method","author":"Oecd","year":""},{"key":"B45","first-page":"442E","volume-title":"In vitro","author":"Oecd","year":""},{"key":"B46","volume-title":"Modern applied statistics with S","author":"Ripley","year":"2002"},{"key":"B47","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1186\/1471-2105-12-77","article-title":"pROC: an open-source package for R and S+ to analyze and compare ROC curves","volume":"12","author":"Robin","year":"2011","journal-title":"BMC Bioinforma."},{"key":"B48","doi-asserted-by":"crossref","DOI":"10.14573\/altex.1411011","volume-title":"Integrated testing strategies (ITS) for safety assessment","author":"Rovida","year":"2015"},{"key":"B49","doi-asserted-by":"publisher","first-page":"774","DOI":"10.1016\/j.tiv.2005.10.014","article-title":"Development of an in vitro skin sensitization test using human cell lines; human Cell Line Activation Test (h-CLAT) II. An inter-laboratory study of the h-CLAT","volume":"20","author":"Sakaguchi","year":"2006","journal-title":"Toxicol. Vitro"},{"key":"B50","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1186\/s12974-015-0247-2","article-title":"Liver X receptor-dependent inhibition of microglial nitric oxide synthase 2","volume":"12","author":"Secor McVoy","year":"2015","journal-title":"J. neuroinflammation"},{"key":"B51","volume-title":"Globally harmonized system of classification and labelling of chemicals (GHS)","author":"Secretariat","year":"2015"},{"key":"B52","doi-asserted-by":"publisher","first-page":"3940","DOI":"10.1093\/bioinformatics\/bti623","article-title":"ROCR: visualizing classifier performance in R","volume":"21","author":"Sing","year":"2005","journal-title":"Bioinformatics"},{"key":"B53","doi-asserted-by":"publisher","first-page":"1150","DOI":"10.1002\/jat.3281","article-title":"Integrated decision strategies for skin sensitization hazard","volume":"36","author":"Strickland","year":"2016","journal-title":"J. Appl. Toxicol."},{"key":"B54","doi-asserted-by":"publisher","first-page":"347","DOI":"10.1002\/jat.3366","article-title":"Multivariate models for prediction of human skin sensitization hazard","volume":"37","author":"Strickland","year":"2017","journal-title":"J. Appl. Toxicol. JAT"},{"key":"B55","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1093\/toxsci\/kfr237","article-title":"An in vitro test to screen skin sensitizers using a stable THP-1\u2013derived IL-8 reporter cell line, THP-G8","volume":"124","author":"Takahashi","year":"2011","journal-title":"Toxicol. Sci."},{"key":"B56","doi-asserted-by":"publisher","first-page":"1318","DOI":"10.1002\/jat.3127","article-title":"Test battery with the human cell line activation test, direct peptide reactivity assay and DEREK based on a 139 chemical data set for predicting skin sensitizing potential and potency of chemicals","volume":"35","author":"Takenouchi","year":"2015","journal-title":"J. Appl. Toxicol."},{"key":"B57","volume-title":"Package \u2018DMwR\u2019","author":"Torgo","year":"2013"},{"key":"B58","doi-asserted-by":"publisher","first-page":"476","DOI":"10.1002\/minf.201000061","article-title":"Best practices for QSAR model development, validation, and exploitation","volume":"29","author":"Tropsha","year":"2010","journal-title":"Mol. Inf."},{"key":"B59","doi-asserted-by":"publisher","first-page":"337","DOI":"10.1016\/j.yrtph.2014.12.008","article-title":"Assessing skin sensitization hazard in mice and men using non-animal test methods","volume":"71","author":"Urbisch","year":"2015","journal-title":"Regul. Toxicol. Pharmacol."},{"key":"B60","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/978-1-4939-6346-1_13","article-title":"Correction of microplate data from high-throughput screening","volume-title":"High-throughput screening assays in Toxicology","author":"Wang","year":"2016"},{"key":"B61","doi-asserted-by":"publisher","first-page":"57","DOI":"10.2174\/1875397301004010057","article-title":"A grid algorithm for high throughput fitting of dose-response curve data","volume":"4","author":"Wang","year":"2010","journal-title":"Curr. Chem. genomics"},{"key":"B62","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1016\/j.toxlet.2020.12.002","article-title":"A direct peptide reactivity assay using a high-throughput mass spectrometry screening platform for detection of skin sensitizers","volume":"338","author":"Wei","year":"2021","journal-title":"Toxicol. Lett."},{"key":"B63","doi-asserted-by":"publisher","first-page":"738","DOI":"10.1080\/10408444.2018.1528207","article-title":"Computational approaches for skin sensitization prediction","volume":"48","author":"Wilm","year":"2018","journal-title":"Crit. Rev. Toxicol."},{"key":"B64","doi-asserted-by":"publisher","first-page":"4833","DOI":"10.3390\/ijms20194833","article-title":"Skin doctor: machine learning models for skin sensitization prediction that provide estimates and indicators of prediction reliability","volume":"20","author":"Wilm","year":"2019","journal-title":"Int. J. Mol. Sci."},{"key":"B65","doi-asserted-by":"publisher","first-page":"846","DOI":"10.1021\/acs.jcim.2c01516","article-title":"Predictive models for human cytochrome P450 3A7 selective inhibitors and substrates","volume":"63","author":"Xu","year":"","journal-title":"J. Chem. Inf. Model."},{"key":"B66","doi-asserted-by":"publisher","first-page":"2321","DOI":"10.1021\/acs.jcim.3c00230","article-title":"Identification of potent and selective acetylcholinesterase\/butyrylcholinesterase inhibitors by virtual screening","volume":"63","author":"Xu","year":"","journal-title":"J. Chem. Inf. Model."},{"key":"B67","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1021\/acs.chemrestox.9b00305","article-title":"Predictive models for human organ toxicity based on in vitro bioactivity data and chemical structure","volume":"33","author":"Xu","year":"2020","journal-title":"Chem. Res. Toxicol."},{"key":"B68","doi-asserted-by":"publisher","first-page":"510","DOI":"10.1021\/ci500667v","article-title":"New publicly available chemical query language, CSRML, to support chemotype representations for application to data mining and modeling","volume":"55","author":"Yang","year":"2015","journal-title":"J. Chem. Inf. Model."},{"key":"B69","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1016\/j.comtox.2019.01.006","article-title":"Automated and standardized workflows in the OECD QSAR Toolbox","volume":"10","author":"Yordanova","year":"2019","journal-title":"Comput. Toxicol."},{"key":"B70","doi-asserted-by":"publisher","first-page":"792","DOI":"10.1002\/jat.3424","article-title":"Prediction of skin sensitization potency using machine learning approaches","volume":"37","author":"Zang","year":"2017","journal-title":"J. Appl. Toxicol. JAT"}],"container-title":["Frontiers in Toxicology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/ftox.2024.1321857\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,2,28]],"date-time":"2024-02-28T05:19:13Z","timestamp":1709097553000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/ftox.2024.1321857\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,28]]},"references-count":70,"alternative-id":["10.3389\/ftox.2024.1321857"],"URL":"https:\/\/doi.org\/10.3389\/ftox.2024.1321857","relation":{},"ISSN":["2673-3080"],"issn-type":[{"value":"2673-3080","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,28]]},"article-number":"1321857"}}