Haematology and Leukaemia - Research Units - Haematology and Leukaemia - Research themes
T cell development and T cell leukaemia/lymphoma
This project entails retroviral transduction of bone marrow or foetal liver with the Ets transcription factor and transplant into lethally irradiated congenic recipients. Disease development is analysed using multiparameter flow cytometry. To identify the leukaemic stem cell or leukaemia-initiating cell, bone marrow or foetal liver subpopulations will be sorted , transduced and transplanted into sublethally irradiated congenic recipients. Initially, these subpopulations will include Lin-c-kit+Sca-1+ cells and common lymphoid progenitors (CLP). The specific mice transplanted with bone marrow or foetal liver subpopulations that develop disease will identify the leukaemic stem cell or leukaemia-initiating cell.
A novel model of mouse myeloid leukaemia
Retroviral transduction of bone marrow or foetal liver with Mixl1 leads to myeloid leukaemia development in transplanted mice. In an effort to identify the leukaemic stem cell in Mixl1-induced myeloid leukaemia, bone marrow or foetal liver subpopulations are being retrovirally transduced with Mixl1. We have retrovirally transduced Lin-c-kit+Sca-1+ cells, common myeloid progenitors, megakaryocyte-erythroid progenitors and granulocyte-macrophage progenitors with Mixl1 and transplanted them into sublethally irradiated recipients. Those mice that develop disease will identify the leukaemic stem cell or leukaemia-initiating cell.
Oncogene discovery using a genetic screen
One double negative (DN) thymocyte can produce one million double positive (DP) thymocytes. We have constructed a cDNA library derived from embryonic day 16 thymocytes which is at a stage of massive cellular expansion. This cDNA library will be retrovirally transduced into bone marrow or foetal liver cells from mice that are deficient in recombinase activation gene 1 (Rag-1). These Rag-1 deficient cells are incapable of rearranging their T cell receptor and as such are blocked at an immature stage of differentiation. cDNA library-transduced Rag-1 deficient cells will be cultured in vitro on stromal cells capable of supporting T cell development. After an appropriate culture time, cells will be analysed by flow cytometry for evidence of traversal of the Rag block in development. If cells are detected that have bypassed the Rag block the rescuing gene/oncogene will be identified using PCR.