Ulrike Künzel, Associate Principal Scientist at AstraZeneca, introduced how AstraZeneca’s arrayed CRISPR screening platform can be used in target identification. By implementing AI and machine learning into the platform, Künzel seeks to tackle challenges associated with primary cells and complex models, including organoids and co-cultures.
AstraZeneca has constructed a framework to improve decision-making and streamline R&D processes. The 5Rs include the right target, right tissue, right safety, right patient, and right commercial potential. This presentation examined the importance of the right target and studying the link between genes and biological pathways that contribute to disease. CRISPR is the gold standard because it enables scientists to specifically knock out one gene at a time and then target the whole genome.
Beyond target identification, Künzel coupled arrayed CRISPR screening with drug treatments and then identified genes that lead to resistance or sensitisation of the drug. This helps to identify biomarkers for patient stratification and new targets for combination therapies.
The team focused on high-content imaging because this can simultaneously assess multiple biomarkers, image the cells, and obtain numerous endpoints and various data points. Künzel also mentioned that automation, including liquid handlers and automated imaging, was critical to achieving reproducible, high-throughput screening. It also minimised variability and enabled adaptation to different primary cell types.
To show this platform in action, Künzel showed how it was applied to chronic kidney disease (CKD), a progressive disease affecting hundreds of millions of people worldwide. The platform was applied to CKD using knowledge graph-derived gene lists. The team screened approximately 250 genes in primary glomerular endothelial cells. Then, they optimised gene editing and assay conditions for robust and scalable results. Künzel added that they opted for nucleofection because cells are difficult to transfect with lipid-based methods.
When the team established an assay for the high-throughput screen, they developed a cytokine stress assay using IL-1β and monitored biomarkers. They were able to identify genes that either prevent or induce stress phenotypes, informing potential therapeutic targets.
Künzel wrapped up her talk by emphasising the significance of democratising access to high-throughput CRISPR screening for academics and SMEs. She announced a new partnership between AstraZeneca, the University of Cambridge, and the Medical Research Council that aims to promote collaboration and wider access to CRISPR screening tools.
