title: Tetraspanin CD9 participates in dysmegakaryopoiesis and stromal interactions in primary myelofibrosis.
abstract: Primary myelofibrosis is characterized by clonal myeloproliferation, dysmegakaryopoiesis, extramedullary hematopoiesis associated with myelofibrosis and altered stroma in the bone marrow and spleen. The expression of CD9, a tetraspanin known to participate in megakaryopoiesis, platelet formation, cell migration and interaction with stroma, is deregulated in patients with primary myelofibrosis and is correlated with stage of myelofibrosis. We investigated whether CD9 participates in the dysmegakaryopoiesis observed in patients and whether it is involved in the altered interplay between megakaryocytes and stromal cells. We found that CD9 expression was modulated during megakaryocyte differentiation in primary myelofibrosis and that cell surface CD9 engagement by antibody ligation improved the dysmegakaryopoiesis by restoring the balance of MAPK and PI3K signaling. When co-cultured on bone marrow mesenchymal stromal cells from patients, megakaryocytes from patients with primary myelofibrosis displayed modified behaviors in terms of adhesion, cell survival and proliferation as compared to megakaryocytes from healthy donors. These modifications were reversed after antibody ligation of cell surface CD9, suggesting the participation of CD9 in the abnormal interplay between primary myelofibrosis megakaryocytes and stroma. Furthermore, silencing of CD9 reduced CXCL12 and CXCR4 expression in primary myelofibrosis megakaryocytes as well as their CXCL12-dependent migration. Collectively, our results indicate that CD9 plays a role in the dysmegakaryopoiesis that occurs in primary myelofibrosis and affects interactions between megakaryocytes and bone marrow stromal cells. These results strengthen the "bad seed in bad soil" hypothesis that we have previously proposed, in which alterations of reciprocal interactions between hematopoietic and stromal cells participate in the pathogenesis of primary myelofibrosis.
authors.affiliation: INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Desterke
authors.affiliation: INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France CTS of Army, Percy Hospital, Clamart, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Martinaud
authors.affiliation: INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France.
authors.forename: Bernadette
authors.initials: B
authors.lastname: Guerton
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Italy.
authors.forename: Lisa
authors.initials: L
authors.lastname: Pieri
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Italy.
authors.forename: Costanza
authors.initials: C
authors.lastname: Bogani
authors.affiliation: INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Clay
authors.affiliation: INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France.
authors.forename: Frederic
authors.initials: F
authors.lastname: Torossian
authors.affiliation: INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France Department of Experimental and Clinical Medicine, University of Florence, Italy.
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Lataillade
authors.affiliation: Department of Hematology, Herlev University Hospital, Copenhagen, Denmark.
authors.forename: Hans C
authors.initials: HC
authors.lastname: Hasselbach
authors.affiliation: Department of Hematology, University Klinik Fur Innere Medizin, Vienna, Austria.
authors.forename: Heinz
authors.initials: H
authors.lastname: Gisslinger
authors.affiliation: Université Catholique de Lille, France French Intergroup on Myeloproliferative Neoplasms (FIM), France.
authors.forename: Jean-Loup
authors.initials: JL
authors.lastname: Demory
authors.affiliation: French Intergroup on Myeloproliferative Neoplasms (FIM), France Department of Hematology, Dr Schaffner Hospital, Lens, France.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Dupriez
authors.affiliation: INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France Inserm U935, Paul Brousse Hospital, Paris-Sud University, Villejuif, France.
authors.forename: Claude
authors.initials: C
authors.lastname: Boucheix
authors.affiliation: INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France Inserm U935, Paul Brousse Hospital, Paris-Sud University, Villejuif, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Rubinstein
authors.affiliation: Department of Hematology, Gustave Roussy Institute, Villejuif, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Amsellem
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Italy.
authors.forename: Alessandro M
authors.initials: AM
authors.lastname: Vannucchi
authors.affiliation: INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France French Intergroup on Myeloproliferative Neoplasms (FIM), France caroline.le-bousse-kerdiles@inserm.fr.
authors.forename: Marie-Caroline
authors.initials: MC
authors.lastname: Le Bousse-Kerdilès
mesh_terms: D018920:Coculture Techniques; D006801:Humans; D008533:Megakaryocytes / Q000378:metabolism* / Q000473:pathology; D055728:Primary Myelofibrosis / Q000378:metabolism* / Q000473:pathology; D017154:Stromal Cells / Q000378:metabolism* / Q000473:pathology; D060245:Tetraspanin 29 / Q000502:physiology*; D034061:Thrombopoiesis / Q000502:physiology*
chemical_list: C554869:CD9 protein, human; D060245:Tetraspanin 29
references.citation: J Biol Chem. 1991 Jan 5;266(1):117-22
references.pmid: 1840589
references.citation: J Biol Chem. 1992 Sep 25;267(27):19665-72
references.pmid: 1527086
references.citation: Br J Haematol. 2001 Jul;114(1):78-83
references.pmid: 11472348
references.citation: Blood. 2001 Apr 1;97(7):1982-9
references.pmid: 11264162
references.citation: J Gastroenterol. 2009;44(9):889-96
references.pmid: 19468669
references.citation: Am J Pathol. 2005 Sep;167(3):849-58
references.pmid: 16127162
references.citation: Leukemia. 1997 Aug;11(8):1290-7
references.pmid: 9264383
references.citation: Platelets. 2011;22(3):196-203
references.pmid: 21231855
references.citation: Leukemia. 2012 May;26(5):1135-7
references.pmid: 22064353
references.citation: Cell Div. 2010 Nov 21;5:27
references.pmid: 21092141
references.citation: Br J Haematol. 2009 Sep;146(5):510-20
references.pmid: 19604240
references.citation: N Engl J Med. 2013 Dec 19;369(25):2391-405
references.pmid: 24325359
references.citation: Blood. 2006 Nov 15;108(10):3472-6
references.pmid: 16868251
references.citation: Leukemia. 2011 Jul;25(7):1217-9
references.pmid: 21537334
references.citation: Leukemia. 2011 Jul;25(7):1200-2
references.pmid: 21455215
references.citation: BMC Cancer. 2011;11:30
references.pmid: 21261996
references.citation: Ann Hematol. 2001 Mar;80(3):160-5
references.pmid: 11320901
references.citation: Blood. 2009 May 28;113(22):5599-604
references.pmid: 19252138
references.citation: Leukemia. 2013 Sep;27(9):1861-9
references.pmid: 23619563
references.citation: Prog Clin Biol Res. 1984;154:427-54
references.pmid: 6089232
references.citation: Leuk Res. 1985;9(5):597-604
references.pmid: 2409408
references.citation: Stem Cells. 2008 Aug;26(8):1920-30
references.pmid: 18511598
references.citation: Blood. 2008 Oct 15;112(8):3026-35
references.pmid: 18669872
references.citation: Exp Hematol. 2008 Dec;36(12):1728-38
references.pmid: 18838203
references.citation: J Cell Sci. 2004 Jul 1;117(Pt 15):3379-88
references.pmid: 15226408
references.citation: Cell Immunol. 1997 Dec 15;182(2):105-12
references.pmid: 9514697
references.citation: Exp Hematol. 2008 Dec;36(12):1585-92
references.pmid: 18814950
references.citation: Blood. 2007 Aug 1;110(3):986-93
references.pmid: 17473062
references.citation: J Clin Pathol. 2009 Apr;62(4):331-8
references.pmid: 19329711
references.citation: Leukemia. 2013 Sep;27(9):1882-90
references.pmid: 23748344
references.citation: Int J Cancer. 2001 May 15;92(4):484-8
references.pmid: 11304681
references.citation: Blood. 2011 Feb 10;117(6):1840-50
references.pmid: 21063023
references.citation: Nucleic Acids Res. 2001 May 1;29(9):e45
references.pmid: 11328886
references.citation: Eur J Immunol. 1996 Nov;26(11):2657-65
references.pmid: 8921952
references.citation: Cancer Res. 2011 Apr 15;71(8):2901-15
references.pmid: 21487043
references.citation: Leukemia. 2010 Jun;24(6):1128-38
references.pmid: 20428194
references.citation: PLoS One. 2011;6(6):e21015
references.pmid: 21698292
references.citation: Leukemia. 2014 Jul;28(7):1407-13
references.pmid: 24441292
references.citation: Acta Biochim Biophys Sin (Shanghai). 2010 Jan;42(1):1-7
references.pmid: 20043041
references.citation: Stem Cells. 2007 Jan;25(1):165-73
references.pmid: 16990584
references.citation: Biochem J. 2009 Jun 1;420(2):133-54
references.pmid: 19426143
references.citation: J Cell Sci. 2006 Feb 15;119(Pt 4):744-52
references.pmid: 16449323
references.citation: Nature. 2005 Apr 28;434(7037):1144-8
references.pmid: 15793561
references.citation: J Cell Physiol. 2000 Jul;184(1):58-69
references.pmid: 10825234
references.citation: Blood. 2002 Aug 15;100(4):1310-7
references.pmid: 12149212