Klin Padiatr 2013; 225 - A11
DOI: 10.1055/s-0033-1343628

Relevance of RUNX1/ETO for Leukaemic Propagation in vivo

N Martinez 1, S Prall 1, H Blair 1, O Heidenreich 1
  • 1NICR, Newcastle University, UK

The t(8;21) translocation generates the RUNX1/ETO fusion protein, which predisposes to acute myeloid leukaemia. However, it is still unclear whether secondary mutations affect its significance for in vivo maintenance of leukaemia. We have addressed this question by using an inducible RUNX1/ETO target knockdown approach in combination with xenotransplantation of immunodeficient mice.

t(8;21) positive SKNO-1 transduced with doxycycline-inducible lentiviral vectors encoding shRNA followed by clonal selection and either competitive cell culture or competitive transplantation into immunodeficient Rag2-/- γc-/- mice with and without doxycycline induction. shRNA compositions of the pools were then analysed by qPCR. Our experiments showed that doxycycline treatment resulted in a loss of RUNX1/ETO shRNA expression. These results confirm previous in vitro experiments using siRNA knockdown and demonstrate for the first time in an in vivo setting the requirement of continuous RUNX1/ETO expression for leukaemic propatagion. Ultimately, these findings further emphasise RUNX1/ETO as a target in t(8;21) positive leukaemia.