Mitochondrial toxicity is an important safety endpoint in drug discovery. Models based solely on chemical structure for predicting mitochondrial toxicity are currently limited in accuracy and applicability domain to the chemical space of the training compounds. In this work, we aimed to utilize both -omics and chemical data to push beyond the state-of-the-art. We combined Cell Painting and Gene Expression data with chemical structural information from Morgan fingerprints for 382 chemical perturbants tested in the Tox21 mitochondrial membrane depolarization assay. We observed that mitochondrial toxicants differ from non-toxic compounds in morphological space and identified compound clusters having similar mechanisms of mitochondrial toxicity, thereby indicating that morphological space provides biological insights related to mechanisms of action of this endpoint. We further showed that models combining Cell Painting, Gene Expression features and Morgan fingerprints improved model performance on an external test set of 244 compounds by 60% (in terms of F1 score) and improved extrapolation to new chemical space. The performance of our combined models was comparable with dedicated in vitro assays for mitochondrial toxicity. Our results suggest that combining chemical descriptors with biological readouts enhances the detection of mitochondrial toxicants, with practical implications in drug discovery.

No abstract

Cell morphology features, such as those from the Cell Painting assay, can be generated at relatively low costs and represent versatile biological descriptors of a system and thereby compound response. In this study, we explored cell morphology descriptors and molecular fingerprints, separately and in combination, for the prediction of cytotoxicity- and proliferation-related in vitro assay endpoints. We selected 135 compounds from the MoleculeNet ToxCast benchmark data set which were annotated with Cell Painting readouts, where the relatively small size of the data set is due to the overlap of required annotations. We trained Random Forest classification models using nested cross-validation and Cell Painting descriptors, Morgan and ErG fingerprints, and their combinations. While using leave-one-cluster-out cross-validation (with clusters based on physicochemical descriptors), models using Cell Painting descriptors achieved higher average performance over all assays (Balanced Accuracy of 0.65, Matthews Correlation Coefficient of 0.28, and AUC-ROC of 0.71) compared to models using ErG fingerprints (BA 0.55, MCC 0.09, and AUC-ROC 0.60) and Morgan fingerprints alone (BA 0.54, MCC 0.06, and AUC-ROC 0.56). While using random shuffle splits, the combination of Cell Painting descriptors with ErG and Morgan fingerprints further improved balanced accuracy on average by 8.9% (in 9 out of 12 assays) and 23.4% (in 8 out of 12 assays) compared to using only ErG and Morgan fingerprints, respectively. Regarding feature importance, Cell Painting descriptors related to nuclei texture, granularity of cells, and cytoplasm as well as cell neighbors and radial distributions were identified to be most contributing, which is plausible given the endpoint considered. We conclude that cell morphological descriptors contain complementary information to molecular fingerprints which can be used to improve the performance of predictive cytotoxicity models, in particular in areas of novel structural space.

The applicability domain of machine learning models trained on structural fingerprints for the prediction of biological endpoints is often limited by the lack of diversity of chemical space of the training data. In this work, we developed similarity-based merger models which combined the outputs of individual models trained on cell morphology (based on Cell Painting) and chemical structure (based on chemical fingerprints) and the structural and morphological similarities of the compounds in the test dataset to compounds in the training dataset. We applied these similarity-based merger models using logistic regression models on the predictions and similarities as features and predicted assay hit calls of 177 assays from ChEMBL, PubChem and the Broad Institute (where the required Cell Painting annotations were available). We found that the similarity-based merger models outperformed other models with an additional 20% assays (79 out of 177 assays) with an AUC > 0.70 compared with 65 out of 177 assays using structural models and 50 out of 177 assays using Cell Painting models. Our results demonstrated that similarity-based merger models combining structure and cell morphology models can more accurately predict a wide range of biological assay outcomes and further expanded the applicability domain by better extrapolating to new structural and morphology spaces. Graphical Abstract

The applicability domain of machine learning models trained on structural fingerprints for the prediction of biological endpoints is often limited by the diversity of chemical space of the training data. In this work, we developed similarity-based merger models which combined the output of individual models trained on cell morphology (based on Cell Painting) and chemical structure (based on chemical fingerprints). Using a combination of a decision tree and logistic regression models on the structural versus morphological feature space of the training data, which leveraged the similarity of test compounds to training compounds, the similarity-based merger models used logistic equations to weigh individual model outputs. We applied these models to predict assay hit calls of 92 assays from ChEMBL and PubChem and 89 anonymised assays released by the Broad Institute, where the required Cell Painting annotations were available. We found that for the 181 assays used in this study the similarity-based merger model improved AUC in relative terms by 16.3% compared to the models using chemical structure alone (mean AUC of 0.75 vs. 0.64), and by 21.3% compared to the models using Cell Painting data alone (mean AUC of 0.62). Our results demonstrate that similarity-based merger models combining structure and cell morphology models can more accurately predict a wide range of biological assay outcomes and expand the applicability domain by better extrapolating to new structural and morphology spaces.