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Open Access 01-12-2012 | Research

Deregulation of apoptosis-related genes is associated with PRV1 overexpression and JAK2 V617F allele burden in Essential Thrombocythemia and Myelofibrosis

Authors: Raquel Tognon, Elainy PL Gasparotto, Renata P Neves, Natália S Nunes, Aline F Ferreira, Patrícia VB Palma, Simone Kashima, Dimas T Covas, Mary Santana, Elizabeth X Souto, Maria Aparecida Zanichelli, Belinda P Simões, Ana Maria de Souza, Fabíola A Castro

Published in: Journal of Hematology & Oncology | Issue 1/2012

Abstract

Background

Essential Thrombocythemia (ET) and Primary Myelofibrosis (PMF) are Chronic Myeloproliferative Neoplasms (MPN) characterized by clonal myeloproliferation/myeloaccumulation without cell maturation impairment. The JAK2 V617F mutation and PRV1 gene overexpression may contribute to MPN physiopathology. We hypothesized that deregulation of the apoptotic machinery may also play a role in the pathogenesis of ET and PMF. In this study we evaluated the apoptosis-related gene and protein expression of BCL2 family members in bone marrow CD34⁺ hematopoietic stem cells (HSC) and peripheral blood leukocytes from ET and PMF patients. We also tested whether the gene expression results were correlated with JAK2 V617F allele burden percentage, PRV1 overexpression, and clinical and laboratory parameters.

Results

By real time PCR assay, we observed that A1, MCL1, BIK and BID, as well as A1, BCLW and BAK gene expression were increased in ET and PMF CD34⁺ cells respectively, while pro-apoptotic BAX and anti-apoptotic BCL2 mRNA levels were found to be lower in ET and PMF CD34⁺ cells respectively, in relation to controls. In patients' leukocytes, we detected an upregulation of anti-apoptotic genes A1, BCL2, BCL-X_L and BCLW. In contrast, pro-apoptotic BID and BIM_EL expression were downregulated in ET leukocytes. Increased BCL-X_L protein expression in PMF leukocytes and decreased BID protein expression in ET leukocytes were observed by Western Blot. In ET leukocytes, we found a correlation between JAK2 V617F allele burden and BAX, BIK and BAD gene expression and between A1, BAX and BIK and PRV1 gene expression. A negative correlation between PRV1 gene expression and platelet count was observed, as well as a positive correlation between PRV1 gene expression and splenomegaly.

Conclusions

Our results suggest the participation of intrinsic apoptosis pathway in the MPN physiopathology. In addition, PRV1 and JAK2 V617F allele burden were linked to deregulation of the apoptotic machinery.

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Tefferi A, Vardiman JW: Classification and diagnosis of myeloproliferative neoplasms: the 2008 World Health Organization criteria and point-of-care diagnostic algorithms. Leukemia. 2008, 22: 14-22. 10.1038/sj.leu.2404955.CrossRefPubMed

Anastasi J: The myeloproliferative neoplasms: insights into molecular pathogenesis and changes in WHO classification and criteria for diagnosis. Hematol Oncol Clin North Am. 2009, 23: 693-708. 10.1016/j.hoc.2009.04.002.CrossRefPubMed

Vainchenker W, Delhommeau F, Constantinescu SN, Bernard OA: New mutations and pathogenesis of myeloproliferative neoplasms. Blood. 2011

James C, Ugo V, Le Couédic JP, Staerk J, Delhommeau F, Lacout C, Garçon L, Raslova H, Berger R, Bennaceur-Griscelli A, Villeval JL, Constantinescu SN, Casadevall N, Vainchenker W: A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature. 2005, 434: 1144-1148. 10.1038/nature03546.CrossRefPubMed

B Baxter EJ, Scott LM, Campbell PJ, East C, Fourouclas N, Swanton S, Vassiliou GS, Bench AJ, Boyd EM, Curtin N, Scott MA, Erber WN, Green AR: Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders. Lancet. 2005, 365: 1054-1061.CrossRef

Levine RL, Wadleigh M, Cools J, Ebert BL, Wernig G, Huntly BJ, Boggon TJ, Wlodarska I, Clark JJ, Moore S, Adelsperger J, Koo S, Lee JC, Gabriel S, Mercher T, D'Andrea A, Fröhling S, Döhner K, Marynen P, Vandenberghe P, Mesa RA, Tefferi A, Griffin JD, Eck MJ, Sellers WR, Meyerson M, Golub TR, Lee SJ, Gilliland DG: Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis. Cancer Cell. 2005, 7: 387-397. 10.1016/j.ccr.2005.03.023.CrossRefPubMed

Kralovics R, Passamonti F, Buser AS, Teo SS, Tiedt R, Passweg JR, Tichelli A, Cazzola M, Skoda RC: A gain-of-function mutation of JAK2 in myeloproliferative disorders. N Engl J Med. 2005, 352: 1779-1790. 10.1056/NEJMoa051113.CrossRefPubMed

Zhao R, Xing S, Li Z, Fu X, Li Q, Krantz SB, Zhao ZJ: Identification of an acquired JAK2 mutation in polycythemia vera. J Biol Chem. 2005, 280: 22788-22792. 10.1074/jbc.C500138200.PubMedCentralCrossRefPubMed

Scott LM, Tong W, Levine RL, Scott MA, Beer PA, Stratton MR, Futreal PA, Erber WN, McMullin MF, Harrison CN, Warren AJ, Gilland DG, Lodish HF, Green AR: JAK2 exon 12 mutations in polycythemia vera and idiopathic erythrocytosis. N Engl J Med. 2007, 356: 459-468. 10.1056/NEJMoa065202.PubMedCentralCrossRefPubMed

10.

Delhommeau F, Dupont S, Della Valle V, James C, Trannoy S, Massé A, Kosmider O, Le Couedic JP, Robert F, Alberdi A, Lécluse Y, Plo I, Dreyfus FJ, Marzac C, Casadevall N, Lacombe C, Romana SP, Dessen P, Soulier J, Viguié F, Fontenay M, Vainchenker W, Bernard AO: Mutation in TET2 in myeloid cancers. N Engl J Med. 2009, 360: 2289-2301. 10.1056/NEJMoa0810069.CrossRefPubMed

11.

Grand FH, Hidalgo-Curtis CE, Ernst T, Zoi K, Zoi C, McGuire C, Kreil S, Jones A, Score J, Metzgeroth G, Oscier D, Hall A, Brandts C, Serve H, Reiter A, Chase AJ, Cross NC: Frequent CBL mutations associated with 11q acquired uniparental disomy in myeloproliferative neoplasms. Blood. 2009, 113: 6182-6192. 10.1182/blood-2008-12-194548.CrossRefPubMed

12.

Pikman Y, Lee BH, Mercher T, McDowell E, Ebert BL, Gozo M, Cuker A, Wernig G, Moore S, Galinsky I, DeAngelo DJ, Clark JJ, Lee SJ, Golub TR, Wadleigh M, Gilliland DG, Levine RL: MPLW515L is a novel somatic activating mutation in myelofibrosis with myeloid metaplasia. PLoS Med. 2006, 3: e270-10.1371/journal.pmed.0030270.PubMedCentralCrossRefPubMed

13.

Carbuccia N, Murati A, Trouplin V, Brecqueville M, Adélaïde J, Rey J, Vainchenker W, Bernard OA, Chaffanet M, Vey N, Birnbaum D, Mozziconacci MJ: Mutations of ASXL1 gene in myeloproliferative neoplasms. Leukemia. 2009, 23: 2183-2186. 10.1038/leu.2009.141.CrossRefPubMed

14.

Larsen TS, Pallisgaard N, Møller MB, Hasselbalch HC: The JAK2 V617F allele burden in essential thrombocythemia, polycythemia vera and primary myelofibrosis--impact on disease phenotype. Eur J Haematol. 2007, 79: 508-515. 10.1111/j.1600-0609.2007.00960.x.CrossRefPubMed

15.

Griesshammer M, Klippel S, Strunck E, Temerinac S, Mohr U, Heimpel H, Pahl HL: PRV-1 mRNA expression discriminates two types of essential thrombocythemia. Ann Hematol. 2004, 83: 364-370. 10.1007/s00277-004-0864-9.CrossRefPubMed

16.

Dillon M, Minear J, Johnson J, Lannutti BJ: Expression of the GPI-anchored receptor Prv-1 enhances thrombopoietin and IL-3-induced proliferation in hematopoietic cell lines. Leuk Res. 2008, 32: 811-819. 10.1016/j.leukres.2007.09.018.CrossRefPubMed

17.

Temerinac S, Klippel S, Strunck E, Röder S, Lübbert M, Lange W, Azemar M, Meinhardt G, Schaefer HE, Pahl HL: Cloning of PRV-1, a novel member of the uPAR receptor superfamily, which is overexpressed in polycythemia rubra vera. Blood. 2000, 95: 2569-2576.PubMed

18.

Klippel S, Strunck E, Temerinac S, Bench AJ, Meinhardt G, Mohr U, Leichtle R, Green AR, Griesshammer M, Heimpel H, Pahl HL: Quantification of PRV-1 mRNA distinguishes polycythemia vera from secondary erythrocytosis. Blood. 2003, 102: 3569-3574. 10.1182/blood-2003-03-0919.CrossRefPubMed

19.

Melis S, Vellinga S, Zachée P, Sierens AC, De Schouwer PJ: JAK2 V617F mutation and PRV-1 overexpression: relevance in the diagnosis of polycythaemia vera and essential thrombocythaemia. Acta Clin Belg. 2009, 64: 429-433.CrossRefPubMed

20.

Zivny J, Klener P, Pytlik R, Andera L: The role of apoptosis in cancer development and treatment: focusing on the development and treatment of hematologic malignancies. Curr Pharm Des. 2010, 16: 11-33. 10.2174/138161210789941883.CrossRefPubMed

21.

Thomadaki H, Scorilas A: BCL2 family of apoptosis-related genes: functions and clinical implications in cancer. Crit Rev Clin Lab Sci. 2006, 43: 1-67. 10.1080/10408360500295626.CrossRefPubMed

22.

Economopoulou C, Pappa V, Kontsioti F, Papageorgiou S, Kapsimali V, Papasteriadi C, Economopoulou P, Papageorgiou E, Dervenoulas J, Economopoulos T: Analysis of apoptosis regulatory genes expression in the bone marrow (BM) of adult de novo myelodysplastic syndromes (MDS). Leuk Res. 2008, 32: 61-69. 10.1016/j.leukres.2007.04.012.CrossRefPubMed

23.

Del Poeta G, Bruno A, Del Principe MI, Venditti A, Maurillo L, Buccisano F, Stasi R, Neri B, Luciano F, Siniscalchi A, de Fabritiis P, Amadori S: Deregulation of the mitochondrial apoptotic machinery and development of molecular targeted drugs in acute myeloid leukemia. Curr Cancer Drug Targets. 2008, 8: 207-222. 10.2174/156800908784293640.CrossRefPubMed

24.

Tognon R, Gasparotto EP, Leroy JM, Oliveira GL, Neves RP, Carrara ReC, Kashima S, Covas DT, Santana M, Souto EX, Zanichelli MA, Velano CE, Simões BP, Alberto FL, Miyashiro K, de Souza AM, Amarante-Mendes GP, de Castro FA: Differential expression of apoptosis-related genes from death receptor pathway in chronic myeloproliferative diseases. J Clin Pathol. 2011, 64: 75-82. 10.1136/jcp.2010.080895.CrossRefPubMed

25.

Mnjoyan Z, Yoon D, Li J, Delhommeau F, Afshar-Kharghan V: The effect of the JAK2 V617F mutation on PRV-1 expression. Haematologica. 2006, 91: 411-412.PubMed

26.

Kralovics R, Teo SS, Buser AS, Brutsche M, Tiedt R, Tichelli A, Passamonti F, Pietra D, Cazzola M, Skoda RC: Altered gene expression in myeloproliferative disorders correlates with activation of signaling by the V617F mutation of Jak2. Blood. 2005, 106: 3374-3376. 10.1182/blood-2005-05-1889.CrossRefPubMed

27.

Olsson A, Norberg M, Okvist A, Derkow K, Choudhury A, Tobin G, Celsing F, Osterborg A, Rosenquist R, Jondal M, Osorio LM: Upregulation of bfl-1 is a potential mechanism of chemoresistance in B-cell chronic lymphocytic leukaemia. Br J Cancer. 2007, 97: 769-777. 10.1038/sj.bjc.6603951.PubMedCentralCrossRefPubMed

28.

Castro FA, Jacysyn JF, Ulbrich AG, Tobo PR, Lopes LR, Colassantti MD, Camanho D, Zanichelli MA, Hamerschlak N, Cavalheiro RC, Oliveira JSR, Amarante-Mendes GP: Overexpression of the Anti-Apoptotic Genes mcl-1, bcl-w, bcl-xL and a1 Is Correlated with Chronic Myelogenous Leukemia Progression and Resistance to Gleevec [abstract]. ASH Annual Meeting Abstracts. 2005, 106: 2880-

29.

Frenzel A, Grespi F, Chmelewskij W, Villunger A: Bcl2 family proteins in carcinogenesis and the treatment of cancer. Apoptosis. 2009, 14: 584-596. 10.1007/s10495-008-0300-z.PubMedCentralCrossRefPubMed

30.

Fabregat I: Dysregulation of apoptosis in hepatocellular carcinoma cells. World J Gastroenterol. 2009, 15: 513-520. 10.3748/wjg.15.513.PubMedCentralCrossRefPubMed

31.

Wuillème-Toumi S, Robillard N, Gomez P, Moreau P, Le Gouill S, Avet-Loiseau H, Harousseau JL, Amiot M, Bataille R: Mcl-1 is overexpressed in multiple myeloma and associated with relapse and shorter survival. Leukemia. 2005, 19: 1248-1252. 10.1038/sj.leu.2403784.CrossRefPubMed

32.

Aichberger KJ, Mayerhofer M, Krauth MT, Skvara H, Florian S, Sonneck K, Akgul C, Derdak S, Pickl WF, Wacheck V, Selzer E, Monia BP, Moriggl R, Valent P, Sillaber C: Identification of mcl-1 as a BCR/ABL-dependent target in chronic myeloid leukemia (CML): evidence for cooperative antileukemic effects of imatinib and mcl-1 antisense oligonucleotides. Blood. 2005, 105: 3303-3311. 10.1182/blood-2004-02-0749.CrossRefPubMed

33.

Eischen CM, Roussel MF, Korsmeyer SJ, Cleveland JL: Bax loss impairs Myc-induced apoptosis and circumvents the selection of p53 mutations during Myc-mediated lymphomagenesis. Mol Cell Biol. 2001, 21: 7653-7662. 10.1128/MCB.21.22.7653-7662.2001.PubMedCentralCrossRefPubMed

34.

Zhang L, Zhao H, Sun A, Lu S, Liu B, Tang F, Feng Y, Zhao L, Yang R, Han ZC: Early down-regulation of Bcl-xL expression during megakaryocytic differentiation of thrombopoietin-induced CD34+ bone marrow cells in essential thrombocythemia. Haematologica. 2004, 89: 1199-1206.PubMed

35.

Mesa RA, Tefferi A, Lasho TS, Loegering D, McClure RF, Powell HL, Dai NT, Steensma DP, Kaufmann SH: Janus kinase 2 (V617F) mutation status, signal transducer and activator of transcription-3 phosphorylation and impaired neutrophil apoptosis in myelofibrosis with myeloid metaplasia. Leukemia. 2006, 20: 1800-1808. 10.1038/sj.leu.2404338.CrossRefPubMed

36.

Rubert J, Qian Z, Andraos R, Guthy DA, Radimerski T: Bim and Mcl-1 exert key roles in regulating JAK2V617F cell survival. BMC Cancer. 2011, 11: 24-10.1186/1471-2407-11-24.PubMedCentralCrossRefPubMed

37.

Chipuk JE, Moldoveanu T, Llambi F, Parsons MJ, Green DR: The BCL-2 family reunion. Mol Cell. 2010, 37: 299-310. 10.1016/j.molcel.2010.01.025.PubMedCentralCrossRefPubMed

38.

Chipuk JE, Green DR: How do BCL-2 proteins induce mitochondrial outer membrane permeabilization?. Trends Cell Biol. 2008, 18: 157-164. 10.1016/j.tcb.2008.01.007.PubMedCentralCrossRefPubMed

39.

Battle TE, Frank DA: The role of STATs in apoptosis. Curr Mol Med. 2002, 2: 381-392. 10.2174/1566524023362456.CrossRefPubMed

40.

Turkson J: STAT proteins as novel targets for cancer drug discovery. Expert Opin Ther Targets. 2004, 8: 409-422. 10.1517/14728222.8.5.409.CrossRefPubMed

41.

Puigdecanet E, Espinet B, Villa O, Florensa L, Besses C, Serrano S, Solé F: Detection of abnormalities of PRV-1, TPO, and c-MPL genes detected by fluorescence in situ hybridization in essential thrombocythemia. Cancer Genet Cytogenet. 2006, 167: 39-42. 10.1016/j.cancergencyto.2005.08.016.CrossRefPubMed

42.

Johansson P, Andréasson B, Safai-Kutti S, Wennström L, Palmqvist L, Ricksten A, Lindstedt G, Kutti J: The presence of a significant association between elevated PRV-1 mRNA expression and low plasma erythropoietin concentration in essential thrombocythaemia. Eur J Haematol. 2003, 70: 358-362. 10.1034/j.1600-0609.2003.00079.x.CrossRefPubMed

Title: Deregulation of apoptosis-related genes is associated with PRV1 overexpression and JAK2 V617F allele burden in Essential Thrombocythemia and Myelofibrosis
Authors: Raquel Tognon
Elainy PL Gasparotto
Renata P Neves
Natália S Nunes
Aline F Ferreira
Patrícia VB Palma
Simone Kashima
Dimas T Covas
Mary Santana
Elizabeth X Souto
Maria Aparecida Zanichelli
Belinda P Simões
Ana Maria de Souza
Fabíola A Castro
Publication date: 01-12-2012
Publisher: BioMed Central
Published in: Journal of Hematology & Oncology / Issue 1/2012
Electronic ISSN: 1756-8722
DOI: https://doi.org/10.1186/1756-8722-5-2

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