Image-based Morphological Profiling Reveals Signatures of Radiation Exposure

ABSTRACT Exposure to ionizing radiation has the potential to induce significant health risks including radiation sickness and death. Here, the utility of image-based morphological profiling (IBMP) assays was investigated as a method to visualize signatures of radiation over time. Human-derived fibroblasts were used as the model system, and were exposed to varying doses of radiation. Cell Painting protocols were then applied to generate images for profiling. Quantitative analysis of images taken from fibroblasts exposed to 1 Gray of ionizing radiation revealed considerable morphological changes by 24 hours post-exposure, with some morphological signs of exposure emerging as early as 4 hours post-exposure. This work demonstrates proof-of-concept for the use of cell painting and IBMP to visualize signatures of radiation, paving the way for its use as a tool for assessing repeated exposures, screening of potential treatments, and establishing relevant timepoints for downstream orthogonal assays. Competing Interest Statement The authors have declared no competing interest. Data availability Processed well-level phenoprints, final cell counts per image, statistical comparisons, and random forest feature information and performance metrics are available in the supplemental materials (Supplementary Datasets S1-S5).