The role of NKX2-1 in T-cell acute lymphoblastic leukemia and NKX2-1 cooperation with the ribosomal RPL10 R98S and NOTCH1 mutations.

T-cell acute lymphoblastic leukemia (T-ALL) is caused by accumulation of at least 10 mutations in immature T-cells. This accumulation does not happen randomly: particular mutations often co-occur while others are never found together. Recent work revealed interesting biology showing how co-occurring mutations actively cooperate in driving leukemia. Ectopic expression of the transcription factor NKX2-1 frequently co-occurs with the ribosomal RPL10 R98S mutation. Whereas we characterized how RPL10 R98S promotes T-ALL and how it cooperates with NOTCH1 mutations, the oncogenic role of NKX2-1 in T-ALL has not been studied. We aim to delineate the role of NKX2-1 in T-ALL and to unravel the molecular mechanisms by which RPL10 R98S and NOTCH1 mutations cooperate with NKX2-1 to promote T-ALL. We will study how NKX2-1 alone or combined with RPL10 R98S and NOTCH1 influences hematopoietic cells in in vitro cell culture and in vivo mouse models. Our preliminary data support co-operation of NKX2-1 and RPL10 R98S at transcriptional level. Therefore, we will identify the direct target genes of NKX2-1 and will test how RPL10 R98S and NOTCH1 affect binding of NKX2-1 to its target genes. Finally, we will perform a targeted CRISPR-Cas9 screen to how dependent leukemia cells are on identified transcriptional changes induced by NKX2-1 + RPL10 R98S + NOTCH1 for their expansion in mice. This CRISPR-Cas9 screen will thus reveal drug targets for this subtype of T-ALL.