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MOLECULAR TARGETS FOR THERAPY

MAPK-negative feedback regulation confers dependence to JAK2V617F signaling

Leukemia volume 37, pages 1686–1697 (2023)Cite this article

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Abstract

Despite significant advances in developing selective JAK2 inhibitors, JAK2 kinase inhibitor (TKI) therapy is ineffective in suppressing the disease. Reactivation of compensatory MEK-ERK and PI3K survival pathways sustained by inflammatory cytokine signaling causes treatment failure. Concomitant inhibition of MAPK pathway and JAK2 signaling showed improved in vivo efficacy compared to JAK2 inhibition alone but lacked clonal selectivity. We hypothesized that cytokine signaling in JAK2V617F induced MPNs increases the apoptotic threshold that causes TKI persistence or refractoriness. Here, we show that JAK2V617F and cytokine signaling converge to induce MAPK negative regulator, DUSP1. Enhanced DUSP1 expression blocks p38 mediated p53 stabilization. Deletion of Dusp1 increases p53 levels in the context of JAK2V617F signaling that causes synthetic lethality to Jak2V617F expressing cells. However, inhibition of Dusp1 by a small molecule inhibitor (BCI) failed to impart Jak2V617F clonal selectivity due to pErk1/2 rebound caused by off-target inhibition of Dusp6. Ectopic expression of Dusp6 and BCI treatment restored clonal selectively and eradicated the Jak2V617F cells. Our study shows that inflammatory cytokines and JAK2V617F signaling converge to induce DUSP1, which downregulates p53 and establishes a higher apoptotic threshold. These data suggest that selectively targeting DUSP1 may provide a curative response in JAK2V617F-driven MPN.

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Fig. 1: Growth factor induced DUSP1 expression confers resistance to ruxolitinib.
Fig. 2: Deletion of Dusp1 is synthetic lethal to JAK2V617F induced MPN.
Fig. 3: Dusp1 deletion eradicates Jak2V617F expressing primary bone marrow cells.
Fig. 4: Chemical inhibition of DUSP1 by BCI suppresses the MPN but lacks clonal selectivity.
Fig. 5: Activation of pERK1/2 due to off-target inhibition of DUSP6 by BCI abrogated its antileukemic response.
Fig. 6: Dusp6 overexpression and BCI treatment eradicated the Jak2V617F clone.

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Data availability

RNA-seq data are publicly available at GSE229318. For materials and other resources, please contact the corresponding author; Mohammad.Azam@cchmc.org.

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Acknowledgements

Authors are thankful to Dr. Benjamin Ebert and Ann Mullaly for providing the Jak2V617F /+ mice and pMSCV-CALRwt and pMSCV-CALRmut retroviral constructs. This study was supported by grants to Mohammad Azam (M Azam) from the National Cancer Institutes at NIH (RO1CA211594) and (RO1CA250516) and to RL by MSKCC Support Grant/Core Grant P30 CA008748. MA is a recipient of the bridge award from the American Society of Hematology (ASH).

Funding

This study was supported by grants to MA from the National Cancer Institutes at NIH (RO1CA211594) and (RO1CA250516) and to RL.L. by MSKCC Support Grant/Core Grant P30 CA008748.

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Authors and Affiliations

  1. Division of Pathology, Cincinnati Children’s Hospital, Cincinnati, OH, USA

    Meenu Kesarwani, Zachary Kincaid, Mohammad Azhar, Jacob Menke, Joshua Schwieterman, Sekhu Ansari, Angela Reaves & Mohammad Azam

  2. Versiti Blood Research Institute and Medical College of Wisconsin, Milwaukee, WI, USA

    Michael E. Deininger

  3. Center for Hematologic Malignancies, and Molecular Cancer Medicine Service, Memorial Sloan-Kettering Cancer Center, New York, NY, USA

    Ross Levine

  4. Division of Immunobiology, Cincinnati Children’s Hospital, Cincinnati, OH, USA

    H. Leighton Grimes

  5. Department of Pediatrics, University of Cincinnati, Cincinnati, OH, USA

    H. Leighton Grimes & Mohammad Azam

  6. Division of Experimental Hematology and Cancer Biology, Cincinnati Children’s Hospital, Cincinnati, OH, USA

    H. Leighton Grimes & Mohammad Azam

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  1. Meenu Kesarwani
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Contributions

MK, ZK, MA, JM, JS, SA, AR, performed the experiments and analyzed the data. MED, RL, and HLG provided patient samples, mouse models, retroviral constructs and guided the experimental design. MK and M Azam designed all experiments. MK and M Azam wrote the manuscript.

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Correspondence to Mohammad Azam.

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Competing interests

MA is a recipient of the bridge award from the American society of hematology (ASH). RLL is on the supervisory board of Qiagen and is a scientific advisor to Imago, Mission Bio, Bakx, Zentalis, Ajax, Auron, Prelude, C4 Therapeutics and Isoplexis. He has received research support from Abbvie, Constellation, Ajax, Zentalis and Prelude. He has received research support from and consulted for Celgene and Roche and has consulted for Syndax, Incyte, Janssen, Astellas, Morphosys and Novartis.

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Kesarwani, M., Kincaid, Z., Azhar, M. et al. MAPK-negative feedback regulation confers dependence to JAK2V617F signaling. Leukemia 37, 1686–1697 (2023). https://doi.org/10.1038/s41375-023-01959-0

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