Lee, J.-Y.JMiller, J.ABasu, SKee, T.-Z.VLoo, L.-HPHARMACOLOGY2020-10-222020-10-222018Lee, J.-Y.J, Miller, J.A, Basu, S, Kee, T.-Z.V, Loo, L.-H (2018). Building predictive in vitro pulmonary toxicity assays using high-throughput imaging and artificial intelligence. Archives of Toxicology 92 (6) : 2055-2075. ScholarBank@NUS Repository. https://doi.org/10.1007/s00204-018-2213-003405761https://scholarbank.nus.edu.sg/handle/10635/179035Human lungs are susceptible to the toxicity induced by soluble xenobiotics. However, the direct cellular effects of many pulmonotoxic chemicals are not always clear, and thus, a general in vitro assay for testing pulmonotoxicity applicable to a wide variety of chemicals is not currently available. Here, we report a study that uses high-throughput imaging and artificial intelligence to build an in vitro pulmonotoxicity assay by automatically comparing and selecting human lung-cell lines and their associated quantitative phenotypic features most predictive of in vivo pulmonotoxicity. This approach is called “High-throughput In vitro Phenotypic Profiling for Toxicity Prediction” (HIPPTox). We found that the resulting assay based on two phenotypic features of a human bronchial epithelial cell line, BEAS-2B, can accurately classify 33 reference chemicals with human pulmonotoxicity information (88.8% balance accuracy, 84.6% sensitivity, and 93.0% specificity). In comparison, the predictivity of a standard cell-viability assay on the same set of chemicals is much lower (77.1% balanced accuracy, 84.6% sensitivity, and 69.5% specificity). We also used the assay to evaluate 17 additional test chemicals with unknown/unclear human pulmonotoxicity, and experimentally confirmed that many of the pulmonotoxic reference and predicted-positive test chemicals induce DNA strand breaks and/or activation of the DNA-damage response (DDR) pathway. Therefore, HIPPTox helps us to uncover these common modes-of-action of pulmonotoxic chemicals. HIPPTox may also be applied to other cell types or models, and accelerate the development of predictive in vitro assays for other cell-type- or organ-specific toxicities. © 2018, The Author(s).Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/amiodaronearistolochic acidbleomycinbutylcresolcadmium chloridecarbamazepineDNAdoxorubicinlithium chloridemyrcenenitrofurantoinnystatinochratoxinparaquatpatulinphalloidinphenylenediamineskatoletenofovirxenobiotic agentArticleartificial intelligenceautomationBEAS-2B cell linecell viability assaychemical structurecontrolled studydiagnostic accuracydiagnostic test accuracy studyDNA damage responseDNA strand breakageHBEC cell line (bronchial epithelium)high throughput in vitro phenotypic profiling for toxicity predictionhigh throughput screeninghumanhuman cellin vitro studyin vivo studylung cell linelung toxicityphenotypepredictive valuepriority journalsensitivity and specificityA-549 cell linebronchuscell linecell survivalchemistrydrug effecthigh throughput screeninglungpathologyprocedurestoxicity testingA549 CellsArtificial IntelligenceBronchiCell LineCell SurvivalHigh-Throughput Screening AssaysHumansLungPredictive Value of TestsSensitivity and SpecificityToxicity TestsXenobioticsArticle