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ACUTE MYELOID LEUKEMIA

Combinatorial genetics reveals the Dock1-Rac2 axis as a potential target for the treatment of NPM1;Cohesin mutated AML

Abstract

Acute myeloid leukemia (AML) is driven by mutations that occur in numerous combinations. A better understanding of how mutations interact with one another to cause disease is critical to developing targeted therapies. Approximately 50% of patients that harbor a common mutation in NPM1 (NPM1cA) also have a mutation in the cohesin complex. As cohesin and Npm1 are known to regulate gene expression, we sought to determine how cohesin mutation alters the transcriptome in the context of NPM1cA. We utilized inducible Npm1cAflox/+ and core cohesin subunit Smc3flox/+ mice to examine AML development. While Npm1cA/+;Smc3Δ/+ mice developed AML with a similar latency and penetrance as Npm1cA/+ mice, RNA-seq suggests that the Npm1cA/+; Smc3Δ/+ mutational combination uniquely alters the transcriptome. We found that the Rac1/2 nucleotide exchange factor Dock1 was specifically upregulated in Npm1cA/+;Smc3Δ/+ HSPCs. Knockdown of Dock1 resulted in decreased growth and adhesion and increased apoptosis only in Npm1cA/+;Smc3Δ/+ AML. Higher Rac activity was also observed in Npm1cA/+;Smc3Δ/+ vs. Npm1cA/+ AMLs. Importantly, the Dock1/Rac pathway is targetable in Npm1cA/+;Smc3Δ/+ AMLs. Our results suggest that Dock1/Rac represents a potential target for the treatment of patients harboring NPM1cA and cohesin mutations and supports the use of combinatorial genetics to identify novel precision oncology targets.

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Fig. 1: Cohesin haploinsufficiency enhances Npm1cA/+ HSPC self-renewal.
Fig. 2: Smc3 haploinsufficiency influences the spectrum of acquired mutations in Npm1cA/+ AML.
Fig. 3: Smc3 haploinsufficiency alters the transcriptional profile of Npm1cA/+ HSPCs.
Fig. 4: Dock1 regulates Npm1cA/+;Smc3Δ/+ AML biology.
Fig. 5: Dock1 functions through Rac2 to regulate apoptosis in Npm1cA/+;Smc3Δ/+ AML cells.
Fig. 6: The Dock1/Rac2 pathway is targetable in Npm1cA/+;Smc3Δ/+ AML.

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Acknowledgements

The authors would like to acknowledge Scott Armstrong and Michael Kühn for providing the OCI-AML3 Cas9 cell line and Benedetta Bonacci for aid with flow cytometry experiments.

Funding

This work was funded by: NCI R01 CA204231 and the Midwest Athletes against Childhood Cancer to SR. AEM is supported by a generous gift from Ms. Nan Gardetto.

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AEM designed research studies, conducted experiments, acquired and analyzed data, and wrote the manuscript. CS conducted experiments and acquired data. KP, RB, QF, and JB analyzed data. ST provided statistical advice and analyzed data. JBF designed research studies, conducted experiments, acquired data, and analyzed data. KEH conducted experiments and acquired data. YZ conducted experiments. ADV and GSV created and provided mouse models for the experiments and provided comments and edits to the manuscript. SR designed research studies and aided in writing the manuscript.

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Correspondence to Sridhar Rao.

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Meyer, A.E., Stelloh, C., Pulakanti, K. et al. Combinatorial genetics reveals the Dock1-Rac2 axis as a potential target for the treatment of NPM1;Cohesin mutated AML. Leukemia 36, 2032–2041 (2022). https://doi.org/10.1038/s41375-022-01632-y

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