Multi-omic Advancement In High-throughput Screening: From Imaging Phenome To Precision Medicine In 2D, 3D, And Beyond

<p dir="ltr">The integration of advanced high content microscopy, sequencing technologies, and other high throughput platforms has created a powerful framework for exploring drug responses in multiomic dimensions. This study presents an integrated approach named <b>MAC-seq</b> (<i>multiplexed analysis of cells</i>) that combines high-throughput imaging with RNA sequencing in 384-well format across various cellular models, including adherent cells, organoids, and advanced 3D co-culture systems (Li et al., 2023). </p><p dir="ltr">Using 2D adherent and 3D spheroid cell models treated with tool compound libraries, we demonstrated how MAC-seq coupled cell painting-based morphological profiling with parallel transcriptomic data. This strategy offers a much deeper understanding of cellular responses, particularly the imaging features derived phenotypic clusters.</p><p dir="ltr">We further adapted this workflow to a high throughput, CAR T cell mediated cytotoxicity assay co-cultured with patient derived tumour spheroids. Live cell imaging captured kinetic differences between CAR-T and untransduced T-cells (UTD). By tracking individual spheroids, the method preserves the tumour heterogeneity and their responses to CAR-T therapy. Furthermore, transcriptomics revealed co-cultured CAR-T cells had distinct biological activity profiles in contrast to CAR-T cells alone or UTD co-culture samples, underscoring the importance of complex culture systems for translational precision medicine.</p><p><br></p><p dir="ltr">Poster presented as part of the Crick BioImage Analysis Symposium 2024.</p><p dir="ltr"><i>Permission has been given by authors to upload to Crick Figshare. Copyright remains with the original authors.</i></p>