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Novel lysyl oxidase inhibitors attenuate hallmarks of primary myelofibrosis in mice

International Journal of Hematology volume 110, pages 699–708 (2019)Cite this article

Abstract

Primary myelofibrosis (PMF) is a chronic myeloproliferative neoplasm (MPN) that usually portends a poor prognosis with limited therapeutic options available. Currently, only allogeneic stem cell transplantation is curative in those who are candidates, while administration of the JAK1/2 inhibitor ruxolitinib carries a risk of worsening cytopenia. The limited therapeutic options available highlight the need for the development of novel treatments for PMF. Lysyl oxidase (LOX), an enzyme vital for collagen cross-linking and extracellular matrix stiffening, has been found to be upregulated in PMF. Herein, we evaluate two novel LOX inhibitors, PXS-LOX_1 and PXS-LOX_2, in two animal models of PMF (GATA1low and JAK2V617F-mutated mice). Specifically, PXS-LOX_1 or vehicle was given to 15- to 16-week-old GATA1low mice via intraperitoneal injection at a dose of 15 mg/kg four times a week for 9 weeks. PXS-LOX_1 was found to significantly decrease the bone marrow fibrotic burden and megakaryocyte number compared to vehicle in both male and female GATA1low mice. Given these results, PXS-LOX_1 was then tested in 15- to 17-week-old JAK2V617F-mutated mice at a dose of 30 mg/kg four times a week for 8 weeks. Again, we observed a significant decrease in bone marrow fibrotic burden. PXS-LOX_2, a LOX inhibitor with improved oral bioavailability, was next evaluated in 15- to 17-week-old JAK2V617F-mutated mice at a dose of 5 mg/kg p.o. four times a week for 8 weeks. This inhibitor also resulted in a significant decrease in bone marrow fibrosis, albeit with a more pronounced amelioration in female mice. Taking these results together, PXS-LOX_1 and PXS-LOX_2 appear to be promising new candidates for the treatment of fibrosis in PMF.

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References

  1. Tefferi A. Myelofibrosis with myeloid metaplasia. N Engl J Med. 2000;342(17):1255–65.

    CAS  Article  Google Scholar 

  2. Visani G, Finelli C, Castelli U, Petti MC, Ricci P, Vianelli N, et al. Myelofibrosis with myeloid metaplasia: clinical and haematological parameters predicting survival in a series of 133 patients. Br J Haematol. 1990;75(1):4–9.

    CAS  Article  Google Scholar 

  3. Tefferi A, Lasho TL, Finke CM, Knudson RA, Ketterling R, Hanson CH, et al. CALR vs JAK2 vs MPL-mutated or triple-negative myelofibrosis: clinical, cytogenetic and molecular comparisons. Leukemia. 2014;28(7):1472–7.

    CAS  Article  Google Scholar 

  4. Tefferi A, Guglielmelli P, Larson DR, Finke C, Wassie EA, Pieri L, et al. Long-term survival and blast transformation in molecularly annotated essential thrombocythemia, polycythemia vera, and myelofibrosis. Blood. 2014;124(16):2507–13 (quiz 2615).

    CAS  Article  Google Scholar 

  5. Zhao R, Xing S, Li Z, Fu X, Li Q, Krantz SB, et al. Identification of an acquired JAK2 mutation in polycythemia vera. J Biol Chem. 2005;280(24):22788–92.

    CAS  Article  Google Scholar 

  6. Klampfl T, Gisslinger H, Harutyunyan AS, Nivarthi H, Rumi E, Milosevic JD, et al. Somatic mutations of calreticulin in myeloproliferative neoplasms. N Engl J Med. 2013;369(25):2379–90.

    CAS  Article  Google Scholar 

  7. Chachoua I, Pecquet C, El-Khoury M, Nivarthi H, Albu RI, Marty C, et al. Thrombopoietin receptor activation by myeloproliferative neoplasm associated calreticulin mutants. Blood. 2016;127(10):1325–35.

    CAS  Article  Google Scholar 

  8. Pardanani AD, Levine RL, Lasho T, Pikman Y, Mesa RA, Wadleigh M, et al. MPL515 mutations in myeloproliferative and other myeloid disorders: a study of 1182 patients. Blood. 2006;108(10):3472–6.

    CAS  Article  Google Scholar 

  9. Leiva O, Ng SK, Chitalia S, Balduini A, Matsuura S, Ravid K. The role of the extracellular matrix in primary myelofibrosis. Blood Cancer J. 2017;7(2):e525.

    CAS  Article  Google Scholar 

  10. Lucero HA, Kagan HM. Lysyl oxidase: an oxidative enzyme and effector of cell function. Cell Mol Life Sci. 2006;63(19–20):2304–16.

    CAS  Article  Google Scholar 

  11. Abbonante V, Chitalia V, Rosti V, Leiva O, Matsuura S, Balduini A, et al. Up-regulation of lysyl oxidase and adhesion to collagen of human megakaryocytes and platelets in primary myelofibrosis. Blood. 2017;130:829–31.

    CAS  Article  Google Scholar 

  12. Eliades A, Papadantonakis N, Bhupatiraju A, Burridge KA, Johnston-Cox HA, Migliaccio AR, et al. Control of megakaryocyte expansion and bone marrow fibrosis by lysyl oxidase. J Biol Chem. 2011;286(31):27630–8.

    CAS  Article  Google Scholar 

  13. Tadmor T, Bejar J, Attias D, Mischenko E, Sabo E, Neufeld G, et al. The expression of lysyl-oxidase gene family members in myeloproliferative neoplasms. Am J Hematol. 2013;88(5):355–8.

    CAS  Article  Google Scholar 

  14. Vannucchi AM, Bianchi L, Cellai C, Paoletti F, Rana RA, Lorenzini R, et al. Development of myelofibrosis in mice genetically impaired for GATA-1 expression (GATA-1(low) mice). Blood. 2002;100(4):1123–32.

    CAS  Article  Google Scholar 

  15. Zingariello M, Sancillo L, Martelli F, Ciaffoni F, Marra M, Varricchio L, et al. The thrombopoietin/MPL axis is activated in the Gata1low mouse model of myelofibrosis and is associated with a defective RPS14 signature. Blood Cancer J. 2017;7(6):e572.

    CAS  Article  Google Scholar 

  16. Lucero HA, Ravid K, Grimsby JL, Rich CB, DiCamillo SJ, Maki JM, et al. Lysyl oxidase oxidizes cell membrane proteins and enhances the chemotactic response of vascular smooth muscle cells. J Biol Chem. 2008;283(35):24103–17.

    CAS  Article  Google Scholar 

  17. Aslam T, Miele A, Chankeshwara S, Megia-Fernandez A, Chesney M, Akram A, McDonald N, Hirani N, Haslett C, Bradley M, Dhaliwal K. Optical molecular imaging of lysyl oxidase activity—detection of active fibrogenesis in human lung tissue. Chem Sci. 2015;6:4946–53.

    CAS  Article  Google Scholar 

  18. Chang J, Lucas MC, Leonte LE, Garcia-Montolio M, Singh LB, Findlay AD, et al. Pre-clinical evaluation of small molecule LOXL2 inhibitors in breast cancer. Oncotarget. 2017;8(16):26066–788.

    Article  Google Scholar 

  19. Schilter H, Findlay AD, Perryman L, Yow TT, Moses J, Zahoor A, et al. The lysyl oxidase like 2/3 enzymatic inhibitor, PXS-5153A, reduces crosslinks and ameliorates fibrosis. J Cell Mol Med. 2018;23:1759–70.

    PubMed  PubMed Central  Google Scholar 

  20. Gilles L, Arslan AD, Marinaccio C, Wen QJ, Arya P, McNulty M, et al. Downregulation of GATA1 drives impaired hematopoiesis in primary myelofibrosis. J Clin Invest. 2017;127(4):1316–20.

    Article  Google Scholar 

  21. Vannucchi AM, Pancrazzi A, Guglielmelli P, Di Lollo S, Bogani C, Baroni G, et al. Abnormalities of GATA-1 in megakaryocytes from patients with idiopathic myelofibrosis. Am J Pathol. 2005;167(3):849–58.

    CAS  Article  Google Scholar 

  22. McDevitt MA, Shivdasani RA, Fujiwara Y, Yang H, Orkin SH. A "knockdown" mutation created by cis-element gene targeting reveals the dependence of erythroid cell maturation on the level of transcription factor GATA-1. Proc Natl Acad Sci USA. 1997;94(13):6781–5.

    CAS  Article  Google Scholar 

  23. Xing S, Wanting TH, Zhao W, Ma J, Wang S, Xu X, et al. Transgenic expression of JAK2V617F causes myeloproliferative disorders in mice. Blood. 2008;111(10):5109–17.

    CAS  Article  Google Scholar 

  24. Matsuura S, Patterson S, Lucero H, Leiva O, Grant AK, Herrera VL, et al. In vivo magnetic resonance imaging of a mouse model of myelofibrosis. Blood Cancer J. 2016;6(11):e497.

    CAS  Article  Google Scholar 

  25. Lucero HA, Patterson S, Matsuura S, Ravid K. Quantitative histological image analyses of reticulin fibers in a myelofibrotic mouse. J Biol Methods. 2016;3(4):e60.

    Article  Google Scholar 

  26. Calapso P, Vitarelli E, Crisafulli C, Tuccari G. Immunocytochemical detection of megakaryocytes by endothelial markers: a comparative study. Pathologica. 1992;84(1090):215–23.

    CAS  PubMed  Google Scholar 

  27. Kroger N, Giorgino T, Scott BL, Ditschkowski M, Alchalby H, Cervantes F, et al. Impact of allogeneic stem cell transplantation on survival of patients less than 65 years of age with primary myelofibrosis. Blood. 2015;125(21):3347–50 (quiz 3364).

    Article  Google Scholar 

  28. Harrison CN, Vannucchi AM, Kiladjian JJ, Al-Ali HK, Gisslinger H, Knoops L, et al. Long-term findings from COMFORT-II, a phase 3 study of ruxolitinib vs best available therapy for myelofibrosis. Leukemia. 2017;31(3):775.

    CAS  Article  Google Scholar 

  29. Verstovsek S, Mesa RA, Gotlib J, Levy RS, Gupta V, DiPersio JF, et al. A double-blind, placebo-controlled trial of ruxolitinib for myelofibrosis. N Engl J Med. 2012;366(9):799–807.

    CAS  Article  Google Scholar 

  30. Tefferi A, Pardanani A. Serious adverse events during ruxolitinib treatment discontinuation in patients with myelofibrosis. Mayo Clin Proc. 2011;86(12):1188–91.

    CAS  Article  Google Scholar 

  31. Arber DA, Orazi A, Hasserjian R, Thiele J, Borowitz MJ, Le Beau MM, et al. The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood. 2016;127(20):2391–405.

    CAS  Article  Google Scholar 

  32. Guglielmelli P, Pacilli A, Rotunno G, Rumi E, Rosti V, Delaini F, et al. Presentation and outcome of patients with 2016 WHO diagnosis of prefibrotic and overt primary myelofibrosis. Blood. 2017;129(24):3227–366.

    CAS  Article  Google Scholar 

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Acknowledgements

This work was supported by a grant from Pharmaxis and by NIHLBI Grant R01HL136363 to KR. SKN was supported by a Cardiovascular Training grant T32 HL007224. OL was recipient of ASH Hematology Opportunities for the Next-Generation of Research Scientists (HONORS) Award from the American Society of Hematology.

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Author notes

  1. Orly Leiva, SengKah Ng and Shinobu Matsuura equally contributed to the study.

Authors and Affiliations

  1. Department of Medicine and Whitaker Cardiovascular Institute, Boston University School of Medicine, 700 Albany St., W-6, Boston, MA, 02118, USA

    Orly Leiva, Seng Kah Ng, Shinobu Matsuura, Hector Lucero & Katya Ravid

  2. Department of Medicine, Brigham and Women’s Hospital, Boston, MA, 02115, USA

    Orly Leiva

  3. Renal Section, Department of Medicine, Boston University School of Medicine, Boston, MA, 02118, USA

    Vipul Chitalia

  4. Pharmaxis Ltd., 20 Rodborough Road, Frenchs Forest, NSW, Australia

    Alison Findlay, Craig Turner & Wolfgang Jarolimek

Authors
  1. Orly Leiva
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  2. Seng Kah Ng
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  3. Shinobu Matsuura
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  4. Vipul Chitalia
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  5. Hector Lucero
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  9. Katya Ravid
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Correspondence to Katya Ravid.

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Conflicts of interests

The authors declare that they have no conflict of interest. The authors affiliated with Pharmaxis synthesized and characterized the inhibitors and tested their efficacy in terms of inhibition of LOX in vivo, but were not involved in performing the experiments related to evaluation of fibrosis burden, and in interpretation of the results.

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Leiva, O., Ng, S.K., Matsuura, S. et al. Novel lysyl oxidase inhibitors attenuate hallmarks of primary myelofibrosis in mice. Int J Hematol 110, 699–708 (2019). https://doi.org/10.1007/s12185-019-02751-6

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  • DOI: https://doi.org/10.1007/s12185-019-02751-6

Keywords

  • Myelofibrosis
  • Lysyl oxidase
  • Myeloproliferative neoplasm