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CHRONIC MYELOPROLIFERATIVE NEOPLASMS

Genetic ablation of Pim1 or pharmacologic inhibition with TP-3654 ameliorates myelofibrosis in murine models

Leukemia volume 36, pages 746–759 (2022)Cite this article

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Abstract

Myelofibrosis (MF) is the deadliest form of myeloproliferative neoplasm (MPN). The JAK inhibitor Ruxolitinib can reduce constitutional symptoms but it does not substantially improve bone marrow fibrosis. Pim1 expression is significantly elevated in MPN/MF hematopoietic progenitors. Here, we show that genetic ablation of Pim1 blocked the development of myelofibrosis induced by Jak2V617F and MPLW515L. Pharmacologic inhibition of Pim1 with a second-generation Pim kinase inhibitor TP-3654 significantly reduced leukocytosis and splenomegaly, and attenuated bone marrow fibrosis in Jak2V617F and MPLW515L mouse models of MF. Combined treatment of TP-3654 and Ruxolitinib resulted in greater reduction of spleen size, normalization of blood leukocyte counts and abrogation of bone marrow fibrosis in murine models of MF. TP-3654 treatment also preferentially inhibited Jak2V617F mutant hematopoietic progenitors in mice. Mechanistically, we show that TP-3654 treatment significantly inhibits mTORC1, MYC and TGF-β signaling in Jak2V617F mutant hematopoietic cells and diminishes the expression of fibrotic markers in the bone marrow. Collectively, our results suggest that Pim1 plays an important role in the pathogenesis of MF, and inhibition of Pim1 with TP-3654 might be useful for treatment of MF.

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Fig. 1: Deletion of Pim1 blocks the development of myelofibrosis in Jak2V617F mouse model.
Fig. 2: Genetic deletion of Pim1 inhibits myelofibrosis in MPLW515L mouse model of MF.
Fig. 3: Pim inhibitor TP-3654 alone or in combination with Ruxolitinib significantly inhibits hematopoietic cells expressing MPN-associated mutants.
Fig. 4: In vivo administration of TP-3654 alone or in combination with Ruxolitinib ameliorates myelofibrosis in Jak2V617F murine model of MF.
Fig. 5: In vivo administration of TP-3654 alone or in combination with Ruxolitinib improves established BM fibrosis in Jak2V617F mouse model.
Fig. 6: TP-3654 alone or in combination with Ruxolitinib treatment preferentially inhibits Jak2V617F mutant hematopoietic progenitors.
Fig. 7: Inhibition of Pim1 by TP-3654 diminishes myelofibrosis in MPLW515L mouse model of MF.
Fig. 8: Suppression of Pim1 activity alters gene expression and cell signaling in MPN cells and attenuates the expression of fibrotic markers.

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Acknowledgements

We thank Dr. Anton Berns of Netherlands Cancer Institute for Pim1 and Pim2 knockout mice. We also thank the Flow Cytometry and Microscopy Core Facilities and the Biorepository and Tissue Research Facility (BTRF) of the University of Virginia for assistance with FACS sorting, confocal microscopy and MPN specimen procurement and processing.

Funding

Flow Cytometry and Microscopy Cores are supported by the UVA Cancer Center through P30CA044578 grant. This work was supported in parts by grants from the Department of Defense (W81XWH1910280) and the National Institutes of Health (R01 HL095685, R01 HL149893) awarded to GM.

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Author notes

  1. These authors contributed equally: Avik Dutta, Dipmoy Nath.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, Charlottesville, VA, USA

    Avik Dutta, Dipmoy Nath, Yue Yang, Bao T. Le, Mohammad Ferdous-Ur Rahman, Patrick Faughnan, Chongzhi Zang & Golam Mohi

  2. Center for Public Health Genomics, University of Virginia School of Medicine, Charlottesville, VA, USA

    Zhenjia Wang, Rongquan He, Wuwei Tan & Chongzhi Zang

  3. Department of Pharmacology, SUNY Upstate Medical University, 750 East Adams Street, Syracuse, NY, USA

    Matthew Stuver

  4. Department of Pathology, SUNY Upstate Medical University, 750 East Adams Street, Syracuse, NY, USA

    Robert E. Hutchison

  5. Sumitomo Dainippon Pharma Oncology, Inc (formerly Tolero Pharmaceuticals, Inc), Lehi, UT, USA

    Jason M. Foulks & Steven L. Warner

  6. Department of Public Health Sciences, University of Virginia School of Medicine, Charlottesville, VA, USA

    Chongzhi Zang

  7. University of Virginia Cancer Center, Charlottesville, VA, USA

    Chongzhi Zang & Golam Mohi

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Contributions

AD performed research, analyzed data and wrote the manuscript; DN performed research and analyzed data; YY performed research; BTL performed data analysis; MFR performed research; PF performed research; ZW, RH, and WT performed RNA-seq data analysis; MS performed research; REH performed histopathologic analysis; JMF and SLW provided TP-3654 and suggestions for research design; CZ supervised RNA-seq data analysis; GM designed the research, analyzed data and wrote the manuscript.

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Correspondence to Golam Mohi.

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Competing interests

G.M. received research funding from Tolero Pharmaceutical Inc. J.M.F. and S.L.W. are employees and stakeholders of Sumitomo Dainippon Pharma Oncology, Inc. (formerly known as Tolero Pharmaceuticals, Inc). The remaining authors declare no competing interests.

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Dutta, A., Nath, D., Yang, Y. et al. Genetic ablation of Pim1 or pharmacologic inhibition with TP-3654 ameliorates myelofibrosis in murine models. Leukemia 36, 746–759 (2022). https://doi.org/10.1038/s41375-021-01464-2

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