Generation and validation of chemosensitisation screens to identify genes interacting with an inhibitor of Menin, a protein with an established role in AML (360G-Wellcome-207070_Z_17_Z)

Dr Vassiliou’s team have recently used the Streptococcus pyogenes-derived type II CRISPR–Cas system to perform genome-wide drop-out screens in acute myeloid leukaemia (AML) cells. Some of the identified AML specific cell-essential genes are being pursued as potential therapeutic targets in AML. The team has adapted this system to perform "chemosensitisation screens" to identify genes whose inhibition collaborates with anti-leukaemic compounds to enhance leukaemia cell kill. I propose to perform chemosensitisation screens to identify genes interacting with an inhibitor of Menin, a protein with an established role in AML. By performing CRISPR-Cas knockout screens in the presence of a cytostatic or low cytotoxic drug doses, genes can be identified whose loss can synergise with the drug. Both resistance (enriched sgRNAs) and sensitization (depleted sgRNAs) genes can be identified by quantitative sequencing of the sgRNA cargo of cells in the presence and absence of the drug. Selected interactions can then be validated using targeted disruption of the targets with sgRNA, an approach that is well-established in the laboratory. Use of chemical inhibitors of target genes, together with the Menin inhibitor, can be used to validate the interaction pharmacologically.