Targeting Myeloma Essential Genes using NOT Gated CAR- T Cells, a computational approach.

Abstract

Sensitive cell surface proteomics studies have shown that the number of completely tumour-specific targets for adoptive cellular immunotherapy is extremely low. Even approved CAR T-cell targets appear to have expression in the central nervous system, leading to long-term neurological complications. We propose that this toxicity could be significantly improved by adoption of NOT-gates, which have been shown to limit CAR T-cell activity against healthy tissue expressing a second target that is absent on the tumour. Furthermore, the approach could also target essential, but non-specific proteins on tumour cells. The use of a NOT gate confers the specificity, whilst targeting the essential protein limits antigen escape. Here we explore the feasibility of such an approach for CAR T-cell targeting of primary myeloma. We show that none of the 45 most essential proteins are unique to the myeloma cell. However, whilst widely expressed, one of the most important proteins for myeloma cell survival, the transferrin receptor, could safely be targeted by a NOT-gate approach. Exploring co-expression patterns demonstrate 26 proteins that are not expressed on myeloma cells, but which are coexpressed with the transferrin receptor in all healthy tissues. We also describe a web app, NOTATER, which can be used by scientists with no bioinformatic capabilities to explore potential NOT-gate combinations in myeloma.