|
|
 Previous Article | Table of Contents | Next Article 
Blood, Vol. 93 No. 1 (January 1), 1999:
pp. 125-139
Autonomous Megakaryocyte Growth in Essential Thrombocythemia and
Idiopathic Myelofibrosis Is Not Related to a c-mpl Mutation
or to an Autocrine Stimulation by Mpl-L
Anne Laure Taksin,
Jean-Pierre Le Couedic,
Isabelle Dusanter-Fourt,
Aline Massé,
Stéphane Giraudier,
André Katz,
Françoise Wendling,
William Vainchenker,
Nicole Casadevall, and
Najet Debili
From INSERM U 362, PR1, Institut Gustave Roussy, Villejuif;
Laboratoire d'hématologie, Hôpital Raymond Poincaré,
Garches; and INSERM U363, ICGM, Hôpital Cochin, Paris, France.
Essential thrombocythemia (ET) and idiopathic myelofibrosis (PMF)
are two myeloproliferative diseases characterized by a marked megakaryocytic (MK) involvement. The pathogenesis of these two diseases
is unknown. Recently it has been shown that overexpression of
Mpl-ligand (Mpl-L) in mice induces thrombocytosis and myelofibrosis. In
this study, we investigated whether Mpl-L was responsible for the
pathogenesis of ET and PMF. Using in vitro cultures of blood or marrow
CD34+ cells, we investigated whether MK growth was
abnormal in these two diseases. Spontaneous MK growth involving only a
fraction (20%) of the MK progenitors, as compared with growth in the
presence of pegylated recombinant human megakaryocyte
growth and development factor (PEG-rhuMGDF), was found in both diseases
(21ET and 14PMF) using serum-free semisolid and liquid cultures,
including cultures at one cell per well. We first searched for a
c-mpl mutation/deletion by sequencing the entire coding region
of the gene by polymerase chain reaction (PCR) in nine ET patients and
five PMF patients, but no mutation was found. We subsequently
investigated whether an autocrine stimulation by Mpl-L could explain
the autonomous MK growth. Addition of different preparations of soluble
Mpl receptor (sMpl) containing a Fc domain of IgG1 (sMpl-Fc) markedly
inhibited MK spontaneous growth in both ET and PMF patients. This
effect was specific for sMpl because a control soluble receptor
(s4-1BB-Fc) had no inhibitory effect and an sMpl devoid of the Fc
fragment had the same inhibitory efficacy as the sMpl-Fc. This
inhibition was reversed by addition of PEG-rhuMGDF or a combination of
cytokines. The sMpl-Fc markedly altered the entry into cell cycle of
the CD34+ cells and increased the apoptosis that occurs
in most patient CD34+ cells in the absence of exogenous
cytokine, suggesting an autocrine stimulation. In contrast, a
neutralizing antibody against Mpl-L did not alter the spontaneous MK
growth, whereas it totally abolished the effects of 10 ng/mL
PEG-rhuMGDF on patient or normal CD34+ cells. Mpl-L
transcripts were detected at a very low level in the patient
CD34+cells and MK and only when a highly sensitive
fluorescent PCR technique was used. By quantitative
reverse-transcription (RT)-PCR, the number of Mpl-L transcripts per
actin transcripts was lower than detected in human Mpl-L-dependent
cell lines, suggesting that this synthesis of Mpl-L was not
biologically significant. In favor of this hypothesis, the Mpl-L
protein was not detected in culture supernatants using either an
enzyme-linked immunosorbent assay (ELISA) or a biological (Ba/F3hu
c-mpl) assay, except in one PMF patient. Investigation of Mpl-L
signaling showed an absence of constitutive activation of STATs in
spontaneously growing patient MKs. Addition of PEG-rhuMGDF to these MKs
activated STATs 3 and 5. This result further suggests that spontaneous
growth is neither related to a stimulation by Mpl-L nor to a
c-mpl mutation. In conclusion, our results show that Mpl-L or
Mpl are not directly implicated in the abnormal proliferation of MK
cells from ET and PMF. The mechanisms by which the sMpl mediates a
growth inhibition will require further experiments.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
A. Jedidi, C. Marty, C. Oligo, L. Jeanson-Leh, J.-A. Ribeil, N. Casadevall, A. Galy, W. Vainchenker, and J.-L. Villeval
Selective reduction of JAK2V617F-dependent cell growth by siRNA/shRNA and its reversal by cytokines
Blood,
August 27, 2009;
114(9):
1842 - 1851.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-J. Lataillade, O. Pierre-Louis, H. C. Hasselbalch, G. Uzan, C. Jasmin, M.-C. Martyre, M.-C. Le Bousse-Kerdiles, and on behalf of the French INSERM and the European EU
Does primary myelofibrosis involve a defective stem cell niche? From concept to evidence
Blood,
October 15, 2008;
112(8):
3026 - 3035.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. A. Beer, P. J. Campbell, L. M. Scott, A. J. Bench, W. N. Erber, D. Bareford, B. S. Wilkins, J. T. Reilly, H. C. Hasselbalch, R. Bowman, et al.
MPL mutations in myeloproliferative disorders: analysis of the PT-1 cohort
Blood,
July 1, 2008;
112(1):
141 - 149.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Lacout, D. F. Pisani, M. Tulliez, F. M. Gachelin, W. Vainchenker, and J.-L. Villeval
JAK2V617F expression in murine hematopoietic cells leads to MPD mimicking human PV with secondary myelofibrosis
Blood,
September 1, 2006;
108(5):
1652 - 1660.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Tefferi
Pathogenesis of Myelofibrosis With Myeloid Metaplasia
J. Clin. Oncol.,
November 20, 2005;
23(33):
8520 - 8530.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. M. Vannucchi, A. Pancrazzi, P. Guglielmelli, S. Di Lollo, C. Bogani, G. Baroni, L. Bianchi, A. R. Migliaccio, A. Bosi, and F. Paoletti
Abnormalities of GATA-1 in Megakaryocytes from Patients with Idiopathic Myelofibrosis
Am. J. Pathol.,
September 1, 2005;
167(3):
849 - 858.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. Vainchenker and S. N. Constantinescu
A Unique Activating Mutation in JAK2 (V617F) Is at the Origin of Polycythemia Vera and Allows a New Classification of Myeloproliferative Diseases
Hematology,
January 1, 2005;
2005(1):
195 - 200.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Tenedini, M. E. Fagioli, N. Vianelli, P. L. Tazzari, F. Ricci, E. Tagliafico, P. Ricci, L. Gugliotta, G. Martinelli, S. Tura, et al.
Gene expression profiling of normal and malignant CD34-derived megakaryocytic cells
Blood,
November 15, 2004;
104(10):
3126 - 3135.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Ding, H. Komatsu, A. Wakita, M. Kato-Uranishi, M. Ito, A. Satoh, K. Tsuboi, M. Nitta, H. Miyazaki, S. Iida, et al.
Familial essential thrombocythemia associated with a dominant-positive activating mutation of the c-MPL gene, which encodes for the receptor for thrombopoietin
Blood,
June 1, 2004;
103(11):
4198 - 4200.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. O'Brien, A. Tefferi, and P. Valent
Chronic Myelogenous Leukemia and Myeloproliferative Disease
Hematology,
January 1, 2004;
2004(1):
146 - 162.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M.-C. Martyre, V. Steunou, M.-C. LeBousse-Kerdiles, J. Wietzerbin, A. M. Vannucchi, and A. R. Migliaccio
Lack of alteration in GATA-1 expression in CD34+ hematopoietic progenitors from patients with idiopathic myelofibrosis
Blood,
June 15, 2003;
101(12):
5087 - 5089.
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Giraudier, H. Chagraoui, E. Komura, S. Barnache, B. Blanchet, J. P. LeCouedic, D. F. Smith, F. Larbret, A.-L. Taksin, F. Moreau-Gachelin, et al.
Overexpression of FKBP51 in idiopathic myelofibrosis regulates the growth factor independence of megakaryocyte progenitors
Blood,
September 26, 2002;
100(8):
2932 - 2940.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. M. Vannucchi, L. Bianchi, C. Cellai, F. Paoletti, R. A. Rana, R. Lorenzini, G. Migliaccio, and A. R. Migliaccio
Development of myelofibrosis in mice genetically impaired for GATA-1 expression (GATA-1low mice)
Blood,
July 30, 2002;
100(4):
1123 - 1132.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. A. Mesa, C. A. Hanson, C.-Y. Li, S.-Y. Yoon, S. V. Rajkumar, G. Schroeder, and A. Tefferi
Diagnostic and prognostic value of bone marrow angiogenesis and megakaryocyte c-Mpl expression in essential thrombocythemia
Blood,
May 13, 2002;
99(11):
4131 - 4137.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Barosi, G. Viarengo, A. Pecci, V. Rosti, G. Piaggio, M. Marchetti, and F. Frassoni
Diagnostic and clinical relevance of the number of circulating CD34+ cells in myelofibrosis with myeloid metaplasia
Blood,
December 1, 2001;
98(12):
3249 - 3255.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. A. Axelrad, D. Eskinazi, P. N. Correa, and D. Amato
Hypersensitivity of circulating progenitor cells to megakaryocyte growth and development factor (PEG-rHu MGDF) in essential thrombocythemia
Blood,
November 15, 2000;
96(10):
3310 - 3321.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Rameshwar, R. Narayanan, J. Qian, T. N. Denny, C. Colon, and P. Gascon
NF-{kappa}B as a Central Mediator in the Induction of TGF-{beta} in Monocytes from Patients with Idiopathic Myelofibrosis: An Inflammatory Response Beyond the Realm of Homeostasis
J. Immunol.,
August 15, 2000;
165(4):
2271 - 2277.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Letestu, N. Vitrat, A. Masse, J.-P. L. Couedic, V. Lazar, P. Rameau, F. Wendling, J. Vuillier, P. Boutard, E. Plouvier, et al.
Existence of a differentiation blockage at the stage of a megakaryocyte precursor in the thrombocytopenia and absent radii (TAR) syndrome
Blood,
March 1, 2000;
95(5):
1633 - 1641.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. R. Moliterno and J. L. Spivak
Posttranslational Processing of the Thrombopoietin Receptor Is Impaired in Polycythemia Vera
Blood,
October 15, 1999;
94(8):
2555 - 2561.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Barosi
Myelofibrosis With Myeloid Metaplasia: Diagnostic Definition and Prognostic Classification for Clinical Studies and Treatment Guidelines
J. Clin. Oncol.,
September 1, 1999;
17(9):
2954 - 2954.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
