Insertional mutagenesis using the Sleeping Beauty transposon system identifies drivers of erythroleukemia in mice

Abstract

Insertional mutagenesis is a powerful means of identifying cancer drivers in animal models. We used the Sleeping Beauty (SB) transposon/transposase system to identify activated oncogenes in hematologic cancers in wild-type mice and mice that express a stabilized cyclin E protein (termed cyclin ET74AT393A). Cyclin E governs cell division and is misregulated in human cancers. Cyclin ET74AT393A mice develop ineffective erythropoiesis that resembles early-stage human myelodysplastic syndrome, and we sought to identify oncogenes that might cooperate with cyclin E hyperactivity in leukemogenesis. SB activation in hematopoietic precursors caused T-cell leukemia/lymphomas (T-ALL) and pure red blood cell erythroleukemias (EL). Analysis of >12,000 SB integration sites revealed markedly different oncogene activations in EL and T-ALL: Notch1 and Ikaros were most common in T-ALL, whereas ETS transcription factors ( Erg and Ets1 ) were targeted in most ELs. Cyclin E status did not impact leukemogenesis or oncogene activations. Whereas most SB insertions were lost during culture of EL cell lines, Erg insertions were retained, indicating Erg’s key role in these neoplasms. Surprisingly, cyclin ET74AT393A conferred growth factor independence and altered Erg-dependent differentiation in EL cell lines. These studies provide new molecular insights into erythroid leukemia and suggest potential therapeutic targets for human leukemia.

Mouse Models of Experimental Glioblastoma

“…In contrast, SB integrations are more randomly distributed, so the PB system can perform more efficient stable gene transfer than the SB system (64)(65)(66). SB and PB transposon systems have been used in both germline and somatic cells of transgenic mice (Figure 3 B, C) (45,60,65,(67)(68)(69)(70)(71).…”

The Evolution of Gene Therapy in the Treatment of Metabolic Liver Diseases

“…Additionally, there have been no reported preclinical SB -associated adverse effects [ 102 , 103 , 104 , 105 ]. The power of SB in identifying cancer driver genes is well known when used in systems with conditional transposase expression in various genetic backgrounds, including a recent murine study that used SB to identify drivers of erythroleukemia in mice [ 106 ]. While intentional insertional mutagenesis is a powerful application of SB , the risk of unintentional insertional mutagenesis appears to be mitigated by the specific conditions under which insertional mutagenesis occurs.…”