Ph-negative chronic myeloproliferative disorders

Pro- and Antifibrotic Factors in the Serum of Patients with Chronic Myeloproliferative Disorders

MYELOID TUMORS , , , ,

AA Silyutina, NM Matyukhina, EG Lisina, VI Khvan, SN Leleko, NT Siordiya, OV Sirotkina, PA Butylin

VA Almazov National Medical Research Center, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341

For correspondence: Pavel Andreevich Butylin, PhD, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341; e-mail: butylinp@gmail.com

For citation: Silyutina AA, Matyukhina NM, Lisina EG, et al. Pro- and Antifibrotic Factors in the Serum of Patients with Chronic Myeloproliferative Disorders. Clinical oncohematology. 2017;10(4):479–84 (In Russ).

DOI: 10.21320/2500-2139-2017-10-4-479-484

ABSTRACT

Background. The study of pro- and antifibrotic factors in the serum of patients with Ph-negative chronic myeloproliferative disorders (CMPDs) will allow to understand better the mechanisms of myelofibrosis development, as well as to identify new diagnostic markers.

Aim. To assess the correlation between the levels of classic (TGF-β, bFGF, MMP-2, -9, -13 and VEGF) and new proinflammatory serum factors (galectin-3), involved into development of myelofibrosis in different Ph-negative forms of CMPDs and genetic abnormalities.

Materials & Methods. The research included 55 CMPD patients (13 with polycythemia vera, 17 with essential thrombocythemia, 25 with primary myelofibrosis) and 8 healthy controls. Whole blood genomic DNA extraction was used to evaluate mutations JAK2V617F, CALR (deletions and insertions), MPLW515L, and MPLW515K. Antibody-immobilized ELISA was used to evaluate the levels of galectin-3, TGF-β, bFGF, VEGF, MMP-2, MMP-9 and MMP-13.

Results. The analysis showed the differences in serum MMP-9, VEGF, TGF-β and galectin-3 levels in patients with different CMPDs. A tendency towards the decrease of serum MMP-9 levels in patients with CALR mutations was shown.

Conclusion. The shown differences between patients with different CMPDs may serve as a basis for improving diagnostic protocols in challenging differential diagnosis of CMPDs.

Keywords: Ph-negative chronic myeloproliferative disorders, pro- and antifibrotic factors, JAK2V617F, CALR, MPLW515L, MPLW515K, MMP-2, MMP-9, MMP-13, galectin-3.

Received: April 26, 2017

Accepted: July 5, 2017

Read in PDF

REFERENCES

  1. Kiladjian J-J. The spectrum of JAK2-positive myeloproliferative neoplasms. Hematology. 2012;2012:561–6. doi: 10.1182/asheducation-2012.1.561.
  2. Nangalia J, Massie CE, Baxter EJ, et al. Somatic CALR mutations in myeloproliferative neoplasms with nonmutated JAK2. N Engl J Med. 2013;369(25):2391–405. doi: 10.1056/NEJMoa1312542.
  3. Klampfl T, Gisslinger H, Harutyunyan AS, et al. Somatic mutations of calreticulin in myeloproliferative neoplasms. N Engl J Med. 2013;369(25):2379–90. doi: 10.1056/NEJMoa1311347.
  4. Li J, Kent DG, Chen E, Green AR. Mouse models of myeloproliferative neoplasms: JAK of all grades. Dis Model Mech. 2011;4(3):311–7. doi: 10.1242/dmm.006817.
  5. Parganas E, Wang D, Stravopodis D, et al. Jak2 is essential for signaling through a variety of cytokine receptors. Cell. 1998;93(3):385–95. doi: 10.1016/s0092-8674(00)81167-8.
  6. Barosi G, Mesa RA, Thiele J, et al. Proposed criteria for the diagnosis of post-polycythemia vera and post-essential thrombocythemia myelofibrosis: a consensus statement from the International Working Group for Myelofibrosis Research and Treatment. Leukemia. 2008;22(2):437–8. doi: 10.1038/sj.leu.2404914.
  7. Hoffman R, Rondelli D. Biology and treatment of primary myelofibrosis. Hematology. 2007;2007(1):346–54. doi: 10.1182/asheducation-2007.1.346.
  8. Jacobson RJ, Salo A, Fialkow PJ. Agnogenic myeloid metaplasia: a clonal proliferation of hematopoietic stem cells with secondary myelofibrosis. Blood. 1978;51(2):189–94.
  9. Reeder TL, Bailey RJ, Dewald GW, et al. Both B and T lymphocytes may be clonally involved in myelofibrosis with myeloid metaplasia. Blood. 2003;101(5):1981–3. doi: 10.1182/blood-2002-07-2341.
  10. Reilly JT. Idiopathic myelofibrosis: pathogenesis, natural history and management. Blood Rev. 1997;11(4):233–42. doi: 10.1016/S0268-960X(97)90022-9.
  11. Mesa RA, Hanson CA, Rajkumar SV, et al. Evaluation and clinical correlations of bone marrow angiogenesis in myelofibrosis with myeloid metaplasia. Blood. 2000;96:3374–80.
  12. Tefferi A. The pathogenesis of chronic myeloproliferative diseases. Int J Hematol. 2001;73(2):170–6. doi: 10.1007/BF02981934.
  13. Chagraoui H, Komura E, Tulliez M, et al. Prominent role of TGF-beta 1 in thrombopoietin-induced myelofibrosis in mice. Blood. 2002;100(10):3495–503. doi: 10.1182/blood-2002-04-1133.
  14. Martyre MC, Le Bousse-Kerdiles C, Romquin N, et al. Elevated levels of basic growth factor in megakaryocytes and platelets from patients with idiopathic myelofibrosis. Br J Haematol. 1997;97(2):441–8. doi: 10.1046/j.1365-2141.1997.292671.x.
  15. Boiocchi L, Vener C, Savi F, et al. Increased expression of vascular endothelial growth factor receptor 1 correlates with VEGF and microvessel density in Philadelphia chromosome-negative myeloproliferative neoplasms. J Clin Pathol. 2011;64(3):226–31. doi: 10.1136/jcp.2010.083386.
  16. Giannandrea M, Parks WC. Diverse functions of matrix metalloproteinases during fibrosis. Dis Mod Mechan. 2014;7(2):193–203. doi: 10.1242/dmm.012062.
  17. Jensen MK, Holten-Andersen MN, Riisbro R, et al. Elevated plasma levels of TIMP-1 correlate with plasma suPAR/uPA in patients with chronic myeloproliferative disorders. Eur J Haematol. 2003;71(5):377–84. doi: 10.1034/j.1600-0609.2003.00096.x.
  18. Wang JC, Novetsky A, Chen C, Novetsky AD. Plasma matrix metalloproteinase and tissue inhibitor of metalloproteinase in patients with agnogenic myeloid metaplasia or idiopathic primary myelofibrosis. Br J Haematol. 2002;119(3):709–12. doi: 10.1046/j.1365-2141.2002.03874.x.
  19. Kim SY, Im K, Park SN, et al. CALR, JAK2, and MPL Mutation Profiles in Patients With Four Different Subtypes of Myeloproliferative Neoplasms: Primary Myelofibrosis, Essential Thrombocythemia, Polycythemia Vera, and Myeloproliferative Neoplasm, Unclassifiable. Am J Clin Pathol. 2015;143(5):635–44. doi: 10.1309/AJCPUAAC16LIWZMM.
  20. Gianelli U, Vener C, Raviele PR, et al. VEGF Expression Correlates With Microvessel Density in Philadelphia Chromosome–Negative Chronic Myeloproliferative Disorders. Am J Clin Pathol. 2007;128(6):966–73. doi: 10.1309/FP0N3LC8MBJUFFA6.
  21. Le Bousse-Kerdiles MC, Chevillard S, Charpentier A, et al. Differential expression of transforming growth factor-beta, basic fibroblast growth factor, and their receptors in CD34+ hematopoietic progenitor cells from patients with myelofibrosis and myeloid metaplasia. Blood. 1996;88:4534–46.
  22. Campanelli R, Rosti V, Villani L, et al. Evaluation of the bioactive and total transforming growth factor β1 levels in primary myelofibrosis. Cytokine. 2011;53(1):100–6. doi: 10.1016/j.cyto.2010.07.427.
  23. Силютина А.А., Гин И.И., Матюхина Н.М. и др. Модели миелофиброза (обзор литературы и собственные данные). Клиническая онкогематология. 2017;10(1):75–84. doi: 10.21320/2500-2139-2017-10-1-75-84.[Silyutina AA, Gin II, Matyukhina NM, et al. Myelofibrosis Models: Literature Review and Own Data. Clinical oncohematology. 2017;10(1):75–84. doi: 10.21320/2500-2139-2017-10-1-75-84. (In Russ)]
  24. Henderson NC, Mackinnon AC, Farnworth SL, et al. Galectin-3 regulates myofibroblast activation and hepatic fibrosis. Proc Nat Acad Sci USA. 2006;103(13):5060–5. doi: 10.1073/pnas.0511167103.
  25. Brand C, Oliveira F, Takiya C, et al. The involvement of the spleen during chronic phase of Schistosoma mansoni infection in galectin-3-/- mice. Histol Histopathol. 2012;27(8):1109–20. doi: 10.14670/HH-27.1109.
  26. Koopmans SM, Bot FJ, Schouten HC, et al. The involvement of Galectins in the modulation of the JAK/STAT pathway in myeloproliferative neoplasia. Am J Blood Res. 2012;2(2):119–27.

Myelofibrosis Models: Literature Review and Own Data

MYELOID TUMORS , , ,

AA Silyutina, II Gin, NM Matyukhina, EN Balayan, PA Butylin

VA Almazov Federal North-West Medical Research Center, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341

For correspondence: Pavel Andreevich Butylin, PhD, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341; e-mail: butylinp@gmail.com

For citation: Silyutina AA, Gin II, Matyukhina NM, et al. Myelofibrosis Models: Literature Review and Own Data. Clinical oncohematology. 2017;10(1):75–84 (In Russ).

DOI: 10.21320/2500-2139-2017-10-1-75-84

ABSTRACT

Background & Aims. Chronic myeloproliferative disorders typically develop during a long latent period, and it complicates the study of the mechanism of its pathogenesis. Observations from the clinical practice should be confirmed by experiments. The mechanisms of oncological transformation related to mutations associated with chronic myeloproliferative diseases were confirmed in transgene animal models. Biological models permitted to determine a complex nature of myelofibrosis. However, studies of the cellular mechanisms of myelofibrosis require new models. This paper presents a review of published models of myeloproliferative disorders, mainly, primary myelofibrosis, and results of studies of a new cell line with expression of JAK2 V617F. The aim of this study is to create a new cell line with expression of transforming JAK2 V617F mutation in acute monocytic leukemia THP-1 cells.

Methods. Transgenic cell lines were created on the basis of monocytic leukemia THP-1 cell line that can differentiate into macrophages. Direct mutagenesis was used to cause V617F mutation. Two cell lines were created: one with JAK2 expression with V617F mutation, the other with wild type JAK2.

Results. Both transgenic lines were characterized by increased JAK2 expression as compared to non-modified cells. In routine cultivation, transgenic THP-1 cells retained the morphology of monocytes. After treatment with phorbol ester, THP-1 differentiated into macrophages and become adherent to culture plastic. Adherent cells demonstrated the variety of shapes: some of them were spherical, the other ones had pseudopodia. No significant differences in viability of cells were observed. However, macrophages expressing mutant JAK2 and overexpressing the wild type JAK2 demonstrated a tendency to decreased amount unlivable cells during cultivation.

Conclusion. The obtained cell model can be used for estimating the influence of JAK2 V617F mutation on pro- and antifibrotic potential of macrophages that can help to investigate the pathogenetic role of macrophages in myelofibrosis development. In addition, this model can help to develop novel methods of therapy and diagnostics of primary and secondary myelofibrosis.

Keywords: Ph-negative chronic myeloproliferative disorders, primary myelofibrosis, JAK2 V617F, transgenic animals.

Received: September 15, 2016

Accepted: December 13, 2016

Read in PDF (RUS)pdficon

REFERENCES

  1. Vardiman JW, Harris NL, Brunning RD. The World Health Organization (WHO) classification of the myeloid neoplasms. Blood 2002;100(7):2292–302. doi: 10.1182/blood-2002-04-1199.
  2. Hoffman R, Rondelli D. Biology and treatment of primary myelofibrosis. Hematol Am Soc Hematol Educ Program. 2007;1:346–54. doi: 10.1182/asheducation-2007.1.346.
  3. Mesa RA, Niblack J, Wadleigh M, et al. The burden of fatigue and quality of life in myeloproliferative disorders (MPDs): an international internet-based survey of 1179 MPD patients. Cancer. 2007;109(1):68–76. doi: 10.1002/cncr.22365.
  4. Cervantes F, Dupriez B, Pereira A, et al. New prognostic scoring system for primary myelofibrosis based on a study of the International Working Group for Myelofibrosis Research and Treatment. Blood. 2009;113(13):2895–901. doi: 10.1182/blood-2008-07-170449.
  5. Scherber R, Dueck AC, Johansson P, et al. The Myeloproliferative Neoplasm Symptom Assessment Form (MPN-SAF): international prospective validation and reliability trial in 402 patients. Blood. 2011;118(2):401–8. doi: 10.1182/blood-2011-01-328955.
  6. Barosi G, Mesa RA, Thiele J, et al. Proposed criteria for the diagnosis of post-polycythemia vera and post-essential thrombocythemia myelofibrosis: a consensus statement from the International Working Group for Myelofibrosis Research and Treatment. Leukemia. 2008;22(2):437–8. doi: 10.1038/sj.leu.2404914.
  7. Tefferi A. Myelofibrosis with myeloid metaplasia. N Engl J Med. 2000;342(17):1255–65. doi: 10.1056/nejm200004273421706.
  8. Jacobson RJ, Salo A, Fialkow PJ. Agnogenic myeloid metaplasia: a clonal proliferation of hematopoietic stem cells with secondary myelofibrosis. Blood. 1978;51:189–94.
  9. Reeder TL, Bailey RJ, Dewald GW, Tefferi A. Both B and T lymphocytes may be clonally involved in myelofibrosis with myeloid metaplasia. Blood. 2003;101(5):1981–3. doi: 10.1182/blood-2002-07-2341.
  10. Reilly JT. Idiopathic myelofibrosis: pathogenesis, natural history and management. Blood Rev. 1997;11(4):233–42. doi: 10.1016/s0268-960x(97)90022-9.
  11. Mehta J, Wang H, Iqbal SU, Mesa R. Epidemiology of myeloproliferative neoplasms (MPN) in the United States. Leuk Lymphoma. 2014;55(3):595–600. doi: 10.3109/10428194.2013.813500.
  12. Kaushansky K. Thrombopoietin: The primary regulator of platelet production. Blood. 1995;86(2):419–31.
  13. de Sauvage FJ, Carver-Moore K, Luoh SM, et al. Physiological regulation of early and late stages of megakaryocytopoiesis by thrombopoietin. J Exp Med. 1996;183(2):651–6.
  14. Vannucchi AM, Villeval J-L, Wagner-Ballon O, et al. Animal Models of Myelofibrosis. In: Conn PM, ed. Sourcebook of Models for Biomedical Research. Totowa: Humana Press Inc.; 2008. pр. 713–23. doi: 10.1007/s12223-008-0071-5.
  15. Ulich TR, del Castillo J, Senaldi G, et al. Systemic hematologic effects of PEG-rHuMGDF-induced megakaryocyte hyperplasia in mice. Blood. 1996;87(12):5006–15.
  16. Villeval JL, Cohen-Solal K, Tulliez M, et al. High thrombopoietin production by hematopoietic cells induces a fatal myeloproliferative syndrome in mice. Blood. 1997;90(11):4369–83.
  17. Yan XQ, Lacey D, Fletcher F, et al. Chronic exposure to retroviral vector encoded MGDF (MPL-ligand) induces lineage-specific growth and differentiation of megakaryocytes in mice. Blood. 1995;86(11):4025–33.
  18. Ohwada A, Rafii S, Moore MA, Crystal RG. In vivo adenovirus vector-mediated transfer of the human thrombopoietin cDNA maintains platelet levels during radiation-and chemotherapy-induced bone marrow suppression. Blood. 1996;88(3):778–84.
  19. Cannizzo SJ, Frey BM, Raffi S, et al. Augmentation of blood platelet levels by intratracheal administration of an adenovirus vector encoding human thrombopoietin cDNA. Nat Biotechnol. 1997;15(6):570–3. doi: 10.1038/nbt0697-570.
  20. Abina MA, Tulliez M, Duffour MT, et al. Thrombopoietin (TPO) knockout phenotype induced by cross-reactive antibodies against TPO following injection of mice with recombinant adenovirus encoding human TPO. J Immunol. 1998;160(9):4481–9.
  21. Frey BM, Rafii S, Teterson M, et al. Adenovector-mediated expression of human thrombopoietin cDNA in immune-compromised mice: Insights into the pathophysiology of osteomyelofibrosis. J Immunol. 1998;160(2):691–9.
  22. Zhou W, Toombs CF, Zou T, et al. Transgenic mice overexpressing human c-MPL ligand exhibit chronic thrombocytosis and display enhanced recovery from 5-fluorouracil or antiplatelet serum treatment. Blood. 1997;89(5):1551–9.
  23. Kakumitsu H, Kamezaki K, Shimoda K, et al. Transgenic mice overexpressing murine thrombopoietin develop myelofibrosis and osteosclerosis. Leuk Res. 2005;29(7):761–9. doi: 10.1016/j.leukres.2004.12.009.
  24. Yanagida M, Ide Y, Imai A, et al. The role of transforming growth factor-beta in PEG-rHuMGDF-induced reversible myelofibrosis in rats. Br J Haematol. 1997;99(4):739–45.
  25. Abina MA, Tulliez M, Lacout C, et al. Major effects of TPO delivered by a single injection of a recombinant adenovirus on prevention of septicemia and anemia associated with myelosuppression in mice: Risk of sustained expression inducing myelofibrosis due to immunosuppression. Gene Ther. 1998;5(4):497–506. doi: 10.1038/sj.gt.3300638.
  26. Shultz LD, SchweitzerPA, Christianson SW, et al. Multiple defects in innate and adaptive immunologic function in NOD/LtSz-SCID mice. J. Immunol. 1995;154:180–91.
  27. Serreze DV, Gaedeke JW, Leiter EH. Hematopoietic stem-cell defects underlying abnormal macrophage development and maturation in NOD/Lt mice: defective regulation of cytokine receptors and protein kinase C. Proc Natl Acad Sci USA. 1993;90(20):9625–9. doi: 10.1073/pnas.90.20.9625.
  28. Wagner-Ballon O, Hedia Chagraoui H, Eric Prina E, et al. Monocyte/Macrophage Dysfunctions Do Not Impair the Promotion of Myelofibrosis by High Levels of Thrombopoietin. J Immunol. 2006;176(11):6425–33. doi: 10.4049/jimmunol.176.11.6425.
  29. Tefferi A. Experimental myelofibrosis in mice and the implications to human disease. Leuk Res. 2005;29(7):723–6. doi: 10.1016/j.leukres.2004.12.006.
  30. Tsai SF, Martin DI, Zon LI, et al. Cloning of cDNA for the major DNA-binding protein of the erythroid lineage through expression in mammalian cells. Nature. 1989;339(6224):446–51. doi: 10.1038/339446a0.
  31. Romeo PH, Prandini MH, Joulin V, et al. Megakaryocytic and erythrocytic lineages share specific transcription factors. Nature. 1990;344(6265):447–9. doi: 10.1038/344447a0.
  32. Yu C, Cantor AB, Yang H, et al. Targeted deletion of a high-affinity GATA-binding site in the GATA-1 promoter leads to selective loss of the eosinophil lineage in vivo. J Exp Med. 2002;195(11):1387–95. doi: 10.1084/jem.20020656.
  33. Migliaccio AR, Rana RA, Sanchez M, et al. GATA-1 as a regulator of mast cell differentiation revealed by the phenotype of the GATA-1low mouse mutant. J Exp Med. 2003;197(3):281–96. doi: 10.1084/jem.20021149.
  34. McDevitt MA, Shivdasani RA, Fujiwara Y, et al. 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. doi: 10.1073/pnas.94.13.6781.
  35. Shivdasani RA, Fujiwara Y, McDevitt MA, et al. A line-age-selective knockout establishes the critical role of transcription factor GATA-1 in megakaryocyte growth and platelet development. EMBO J. 1997;16(13):3965–73. doi: 10.1093/emboj/16.13.3965.
  36. Vyas P, Ault K, Jackson CW, et al. Consequences of GATA-1 deficiency in megakaryocytes and platelets. Blood. 1999;93(9):2867–75.
  37. Centurione L, Di Baldassarre A, Zingariello M, et al. Increased and pathologic emperipolesis of neutrophils within megakaryocytes associated with marrow fibrosis in GATA-1(low) mice. Blood. 2004;104(12):3573–80. doi: 10.1182/blood-2004-01-0193.
  38. Vannucchi AM, Bianchi L, Cellai C, et al. Development of myelofibrosis in mice genetically impaired for GATA-1 expression (GATA-1(low) mice). Blood. 2002;100(4):1123–32. doi: 10.1182/blood-2002-06-1913.
  39. Vannucchi AM, Migliaccio AR, Paoletti F, et al. Pathogenesis of myelofibrosis with myeloid metaplasia: Lessons from mouse models of the disease. Semin Oncol. 2005;32(4):365–72. doi: 10.1053/j.seminoncol.2005.04.008.
  40. Radich J. The Molecular Biology of Myeloproliferative Disorders. Cancer Cell. 2010;18(1):7–8. doi: 10.1016/j.ccr.2010.06.006.
  41. Baxter EJ, Scott LM, Campbell PJ, et al. Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders. Lancet. 2005;365(9464):1054–61. doi: 10.1016/s0140-6736(05)74230-6.
  42. Kralovics R, Passamonti F, Buser AS, et al. A gain-of-function mutation of JAK2 in myeloproliferative disorders. N Engl J Med. 2005;352(17):1779–90. doi: 10.1056/nejmoa051113.
  43. Levine RL, Wadleigh M, Cools J, et al. Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis. Cancer Cell. 2005;7(4):387–97. doi: 10.1016/j.ccr.2005.03.023.
  44. Kiladjian JJ. The spectrum of JAK2-positive myeloproliferative neoplasms. Hematol Am Soc Hematol Educ Program. 2012;2012:561–6. doi: 10.1182/asheducation-2012.1.561.
  45. Levine RL, Gilliland DG. Myeloproliferative disorders. Blood. 2008;112(6):2190–8. doi: 10.1182/blood-2008-03-077966.
  46. Chen M, Cheng A, Chen YQ, et al. The amino terminus of JAK3 is necessary and sufficient for binding to the common gamma chain and confers the ability to transmit interleukin 2-mediated signals. Proc Natl Acad Sci USA. 1997;94(13):6910–5. doi: 10.1073/pnas.94.13.6910.
  47. Saharinen P, Silvennoinen O. The pseudokinase domain is required for suppression of basal activity of Jak2 and Jak3 tyrosine kinases and for cytokine-inducible activation of signal transduction. J Biol Chem. 2002;277(49):47954–63. doi: 10.1074/jbc.m205156200.
  48. Griffith J, Black J, Faerman C, et al. The structural basis for autoinhibition of FLT3 by the juxtamembrane domain. Mol Cell. 2004;13(2):169–78. doi: 10.1016/s1097-2765(03)00505-7.
  49. Lindauer K, Loerting T, Liedl KR, et al. Prediction of the structure of human Janus kinase 2 (JAK2) comprising the two carboxy-terminal domains reveals a mechanism for autoregulation. Protein Engin. Design Select. 2001;14(1):27–37. doi: 10.1093/protein/14.1.27.
  50. Parganas E, Wang D, Stravopodis D, et al. Jak2 is essential for signaling through a variety of cytokine receptors. Cell. 1998;93(3):385–95. doi: 10.1016/s0092-8674(00)81167-8.
  51. Levine RL, Loriaux M, Huntly BJ, et al. The JAK2V617F activating mutation occurs in chronic myelomonocytic leukemia and acute myeloid leukemia, but not in acute lymphoblastic leukemia or chronic lymphocytic leukemia. Blood. 2005;106(10):3377–9. doi: 10.1182/blood-2005-05-1898.
  52. Jelinek J, Oki Y, Gharibyan V, et al. JAK2 mutation 1849G>T is rare in acute leukemias but can be found in CMML, Philadelphia-chromosome negative CML and megakaryocytic leukemia. Blood. 2005;106(10):3370–3. doi: 10.1182/blood-2005-05-1800.
  53. Steensma DP, Dewald GW, Lasho TL, et al. The JAK2 V617F activating tyrosine kinase mutation is an infrequent event in both “atypical” myeloproliferative disorders and the myelodysplastic syndrome. Blood. 2005;106(4):1207–9. doi: 10.1182/blood-2005-03-1183.
  54. James C, Ugo V, Le Couedic JP, et al. A unique clonal JAK2 mutation leading to constitutive signaling causes polycythaemia vera. Nature. 2005;434(7037):1144–8. doi: 10.1038/nature03546.
  55. Scott LM, Scott MA, Campbell PJ, Green AR. Progenitors homozygous for the V617F mutation occur in most patients with polycythemia vera, but not essential thrombocythemia. Blood. 2006;108(7):2435–7. doi: 10.1182/blood-2006-04-018259.
  56. Wernig G, Mercher T, Okabe R, et al. Expression of Jak2V617F causes a polycythemia vera-like disease with associated myelofibrosis in a murine bone marrow transplant model. Blood. 2006;107(11):4274–81. doi: 10.1182/blood-2005-12-4824.
  57. Lacout C, Pisani DF, Tulliez M, et al. JAK2V617F expression in murine hematopoietic cells leads to MPD mimicking human PV with secondary myelofibrosis. Blood. 2006;108(5):1652–60. doi: 10.1182/blood-2006-02-002030.
  58. Tiedt R, Hao-Shen H, Sobas MA, et al. Ratio of mutant JAK2-V617F to wild type JAK2 determines the MPD phenotypes in transgenic mice. Blood. 2007;111(8):3931–40. doi: 10.1182/blood-2007-08-107748.
  59. Xing S, Ho WT, Zhao W, et al. Transgenic expression of JAK2V617F causes myeloproliferative disorders in mice. Blood. 2008;111(10):5109–17. doi: 10.1182/blood-2007-05-091579.
  60. Marty C, Lacout C, Martin A, et al. Myeloproliferative neoplasm induced by constitutive expression of JAK2V617F in knock-in mice. Blood. 2010;116(5):783–7. doi: 10.1182/blood-2009-12-257063.
  61. Li J, Kent DG, Chen E, Green AR. Mouse models of myeloproliferative neoplasms: JAK of all grades. Dis Model Mech. 2011;4(3):311–7. doi: 10.1242/dmm.006817.
  62. Pikman Y, Lee BH, Mercher T, et al. MPLW515L is a novel somatic activating mutation in myelofibrosis with myeloid metaplasia. PLoS Med. 2006;3(7):e270. doi: 10.1371/journal.pmed.0030270.
  63. Pardanani AD, Levine RL, Lasho T, et al. MPL515 mutations in myeloproliferative and other myeloid disorders: A study of 1182 patients. Blood. 2006;108(10):3472–6. doi: 10.1182/blood-2006-04-018879.
  64. Nangalia J, Massie CE, Baxter EJ, et al. Somatic CALR mutations in myeloproliferative neoplasms with nonmutated JAK2. N Engl J Med. 2013;369(25):2391–405. doi: 10.1056/NEJMoa1312542.
  65. Klampfl T, Gisslinger H, Harutyunyan AS, et al. Somatic mutations of calreticulin in myeloproliferative neoplasms. N Engl J Med. 2013;369(25):2379–90. doi: 10.1056/NEJMoa1311347.
  66. Wang WA, Groenendyk J, Michalak M. Calreticulin signaling in health and disease. Int J Biochem Cell Biol. 2012;44(6):842–6. doi: 10.1016/j.biocel.2012.02.009.
  67. Michalak M, Groenendyk J, Szabo E, et al. Calreticulin, a multi-process calcium-buffering chaperone of the endoplasmic reticulum. Biochem J. 2009;417(3):651–66. doi: 10.1042/BJ20081847.
  68. Gold LI, Eggleton P, Sweetwyne MT, et al. Calreticulin: non-endoplasmic reticulum functions in physiology and disease. FASEB J. 2010;24(3):665–83. doi: 10.1096/fj.09-145482.
  69. Luo B, Lee AS. The critical roles of endoplasmic reticulum chaperones and unfolded protein response in tumorigenesis and anticancer therapies. Oncogene. 2012;32(7):805–18. doi: 10.1038/onc.2012.130.
  70. Rumi E, Pietra D, Ferretti V, et al. JAK2 or CALR mutation status defines subtypes of essential thrombocythemia with substantially different clinical course and outcomes. Blood. 2014;123(10):1544–51. doi: 10.1182/blood-2013-11-539098.
  71. Cabagnols X, Defour JP, Ugo V, et al. Differential association of calreticulin type 1 and type 2 mutations with myelofibrosis and essential thrombocytemia: relevance for disease evolution. Leukemia. 2014;29(1):249–52. doi: 10.1038/leu.2014.270.
  72. Marty C, Pecquet C, Nivarthi H, et al. Calreticulin mutants in mice induce an MPL-dependent thrombocytosis with frequent progression to myelofibrosis. Blood. 2016;127(10):1317–24. doi: 10.1182/blood-2015-11-679571.
  73. Chachoua I, Pecquet C, El-Khoury M, et al. Thrombopoietin receptor activation by myeloproliferative neoplasm associated calreticulin mutants. Blood. 2016;127(10):1325–35. doi: 10.1182/blood-2015-11-681932.
  74. Thiele J, Kvasnicka HM, Boeltken B. Resident bone marrow macrophages in idiopathic (primary) myelofibrosis (IMF): a histochemical and morphometric study on sequential trephine biopsies. Leuk Res. 1999;23(11):983–5. doi: 10.1016/s0145-2126(99)00120-4.
  75. Tsuchiya S, Kobayashi Y, Goto Y, et al. Induction of maturation in cultured human monocytic leukemia cells by a phorbol diester. Cancer Res. 1982;42(4):1530–6.