Precise single-cell transcriptomic mapping of normal and leukemic cell states reveals unconventional lineage priming in acute myeloid leukemia

Zeng, A. G. X.; Iacobucci, I.; Shah, S.; Mitchell, A.; Wong, G.; Bansal, S.; Gao, Q.; Kim, H.; Kennedy, J. A.; Minden, M. D.; Haferlach, T.; Mullighan, C. G.; Dick, J. E.

doi:10.1101/2023.12.26.573390

bioRxiv · 10.1101/2023.12.26.573390

Precise single-cell transcriptomic mapping of normal and leukemic cell states reveals unconventional lineage priming in acute myeloid leukemia

Zeng, A. G. X.·Iacobucci, I.·Shah, S.·Mitchell, A.·Wong, G.·Bansal, S.·Gao, Q.·Kim, H.·Kennedy, J. A.·Minden, M. D.·Haferlach, T.·Mullighan, C. G.·Dick, J. E.

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Abstract

In acute myeloid leukemia (AML), genetic mutations distort hematopoietic differentiation, resulting in the accumulation of leukemic blasts. Yet, it remains unclear how these mutations intersect with cellular origins and whether they converge upon similar differentiation patterns. Single-cell RNA sequencing (scRNA-seq) has enabled high-resolution mapping of the relationship between leukemia and normal cell states, yet this application is hampered by imprecise reference maps of normal hematopoiesis and small sample sizes among patient cohorts. As a first step we constructed a reference atlas of human bone marrow hematopoiesis from 263,519 single-cell transcriptomes spanning 55 cellular states, that was benchmarked against independent datasets of immunophenotypically pure hematopoietic stem and progenitor cells. Using this reference atlas, we mapped over 1.2 million single-cell transcriptomes spanning 318 AML, mixed phenotype acute leukemia (MPAL), and acute erythroid leukemia (AEL) samples. This large-scale analysis, together with systematic mapping of genotype-to-phenotype associations between driver mutations and differentiation landscapes, revealed convergence of diverse genetic alterations on twelve recurrent patterns of aberrant differentiation in AML. This included unconventional lymphoid and erythroid priming linked to RUNX1 and TP53 mutations, respectively. We also identified non-genetic determinants of AML differentiation such as two subgroups of KMT2A-rearranged AML that differ in the identity of their leukemic stem cells (LSCs), likely reflecting distinct cellular origins. Furthermore, distinct LSC-driven hierarchies can co-exist within individual patients, providing insights into AML evolution. Together, precise mapping of normal and malignant cell states provides a framework for advancing the study and disease classification of hematologic malignancies thereby informing therapy development.

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Zeng, A. G. X., Iacobucci, I., Shah, S., Mitchell, A., Wong, G., Bansal, S., Gao, Q., Kim, H., Kennedy, J. A., Minden, M. D., Haferlach, T., Mullighan, C. G., Dick, J. E.. 2023-12-27. Precise single-cell transcriptomic mapping of normal and leukemic cell states reveals unconventional lineage priming in acute myeloid leukemia. https://doi.org/10.1101/2023.12.26.573390

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