Loss_of__Driver__Transposon_Integrations_in_Acute_Myeloid_Leukaemia__

Analysis of transposon integration sites in serial blood samples from mice with the Npm1cA mutation has shown only a small proportion of transposon integrations occur early and persist through serial blood samples and on serial transplantation of leukaemia cells to NSG mice. These are the putative tumour drivers. Integrations in the NF1 gene are seen frequently in this cohort and persist through several of the pre-leukaemic blood samples, but are often lost from the final tumour and transplant samples. There are several possible explanations. These Nf1 integrations may be occuring in a different sub-clone to the dominant clone or the SB footprint may be having a mutagenic effect after the transposon has remobilised. A third explanation is that these integrationshave a role in initiating the tumour clone, but once additional driver integrations accumulate within the cell the Nf1 integration is no longer required for tumour cell proliferation and survival. To investigate these possibilities, we will screen for the Sleeping Beauty transposon footprint in regions of the Nf1 gene where transposon integrations were lost, initially in whole tumour DNA. If these can be detected by the MiSeq approach we will screen colonies derived from single haematopoietic cells for the relevant footprint co-occuring with driver mutations at the single cell level.