2008 – INTEGRATIVE SINGLE-CELL ANALYSIS OF PRELEUKAEMIC MUTANT MOUSE MODELS ILLUSTRATES MUTATION-SPECIFIC HAEMATOPOIETIC PERTURBATIONS

Acute myeloid leukaemia can develop from antecedent conditions such as myeloproliferative neoplasms as well as clonal haematopoiesis through stepwise acquisition of multiple mutations. However, the perturbing effects of individual mutations on the entire haematopoietic system remain elusive. To better characterise the mutation-specific haematopoietic disruption, we conducted single-cell RNA sequencing of eight different mutant mouse models with the following mutations: Flt3 internal tandem duplication (n=3), Npm1c (n=2), Idh1 R132H (n=3), Dnmt3a R882H (n=2), Ezh2 knock-out (n=2), Utx knock-out (n=2), Jak2 V617F (n=2) and Calr 52-bp deletion (n=2).Compared with the wild-type samples, each mutant model showed significant accumulation of distinct subsets of progenitors: granulomonocytic progenitors in the Flt3 mutant, stem/immature cells in the Idh1 mutant, lymphoid and erythroid progenitors in the Ezh2 knock-out, granulocyte progenitors and megakaryocytes in the Utx mutant, late erythroid progenitors in the Jak2 mutant, and stem cells and megakaryocytes in the Calr mutant. Npm1 and Dnmt3a mutants showed smaller effects on the global cell type distribution. Transcriptome-based inference of cellular metabolism further revealed that energy generating glycolysis and TCA-cycle reactions were differentially associated with the cell type abundance, where Jak2 and Calr mutations activated these metabolic processes in the accumulated stages of progenitors, while Idh1, Ezh2 and Utx mutations downregulated these energy generation processes in the abundant cell types.I will present an integrated analysis of eight preleukemic mouse models, and the different methods which I have utilised to investigate differential abundance of cell populations, metabolic analysis, differential gene expression and differentiation biases of the progenitor cell populations.