Transcriptomic characterization of murine Trp53-mutant acute erytroid leukemias

We established a mouse model to study the development of therapy-related myeloid neoplasms (t-MN) arising from TP53-mutant clonal hematopoiesis. Mice carrying conditional Trp53-fl-R245W-GFP alleles, in combination with the inducible hematopoietic-specific Cre driver (SCL-CreERT), allow induction of either monoallelic or biallelic Trp53 missense mutations. To mimic cytotoxic therapies implicated in t-MN pathogenesis, mice were exposed to sublethal γ-irradiation, which promoted the development of hematologic malignancies, including acute erythroid leukemia. We then subjected these leukemias to bulk RNA sequencing to characterize their transcriptomic landscape. We FACS-purified leukemic blasts from mice with mono- or biallelic Trp53-fl-R245W-GFP alleles, and performed bulk RNA sequencing to define transcriptomic changes associated with malignant transformation. As controls, healthy megakaryocyte-erythroid progenitors (MEPs), immunophenotypically resembling the AML blasts, from mice with biallelic Trp53-fl-R245W-GFP alleles were sorted into GFP negative (Trp53 wild-type), GFP low (monoallelic Trp53 mutation) and GFP high (biallelic Trp53 mutation) populations.