Physical Poster + E-Poster Presentation 34th Lorne Cancer Conference 2022

Oncogenic Notch1 can replace the Lmo2/Lyl1 stem cell program in murine Lmo2-induced T-cell acute lymphoblastic leukemia (#126)

Sung K Chiu 1 , James The 2 , Jesslyn Saw 2 , Jacqueline A Boyle 2 , Feng Yan 2 , Ralf B Schittenhelm 3 , Anh NQ Vo 2 , Benjamin J Shields 2 , Anna L Leichter 2 , Matthew P McCormack 2 , Cedric S Tremblay 2 , David J Curtis 2 4
  1. Telethon Kids Institute, University of WA, Perth, WA, Australia
  2. Australian Centre for Blood Diseases, Monash University, Melbourne, VIC, Australia
  3. Monash Proteomics & Metabolomics Facility, Monash University, Melbourne, Victoria, Australia
  4. Clinical Haematology, Alfred Health, Melbourne, VIC, Australia

Co-expression of LMO2 and LYL1 typifies the high-risk early thymocyte precursor subtype of T-cell acute lymphoblastic leukemia (T-ALL), which has a stem cell-like immunophenotype. Using a Lmo2 transgenic mouse, we have shown previously that T-cell expression of Lmo2 establishes a pool of immature thymocytes (pre-LSCs) with stem cell-like properties including self-renewal, quiescence and chemo-resistance. To understand how Lmo2 induces this stem cell phenotype, mass spectrometry of mouse and human T-ALL cells identified a core complex including Lmo2, Ldb1, Runx1, Cbfa2t3, Lyl1 and Tal1, a close relative of Lyl1. ChIP-seq and ATAC-seq showed this complex bound regulatory elements of the heptad of transcription factors (Lyl1, Fli-1, Erg, Gata2 but not Tal1) that regulate normal hematopoietic stem and progenitor cells (HSPCs). To determine if leukemic cells remained addicted to this Lmo2/Lyl1 stem cell-like program, we generated a conditional Lyl1 knockout mouse strain to enable deletion of Lyl1 in pre-LSCs or T-ALL. At the pre-LSC stage, deletion of Lyl1 led to loss of self-renewal and reduced expression of Hhex, a direct target of the Lmo2/Lyl1 complex. Deletion of Lyl1 in T-ALL also reduced Hhex expression, but leukemic cell growth continued with expansion of Notch1-mutated clones. Expression of an oncogenic Notch1 transgene rescued self-renewal of Lyl1-deleted pre-LSCs. Expression analysis of the human LMO2/LYL1 subgroup of T-ALL identified some cases with low HHEX and elevated DTX, a target of NOTCH signaling. Thus, Lmo2 induces a stem cell-like network similar to normal HSPCs but this program can be supplanted by oncogenic Notch1 signaling during clonal evolution.