Tetraspanin CD9 participates in dysmegakaryopoiesis and stromal interactions in primary myelofibrosis
Christophe Desterke,
Christophe Martinaud,
Bernadette Guerton,
Lisa Pieri,
Costanza Bogani,
Denis Clay,
Frederic Torossian,
Jean-Jacques Lataillade,
Hans C. Hasselbach,
Heinz Gisslinger,
Jean-Loup Demory,
Brigitte Dupriez,
Claude Boucheix,
Eric Rubinstein,
Sophie Amsellem,
Alessandro M. Vannucchi,
Marie-Caroline Le Bousse-Kerdilès
Affiliations
Christophe Desterke
INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France;INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France
Christophe Martinaud
INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France;CTS of Army, Percy Hospital, Clamart, France
Bernadette Guerton
INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France
Lisa Pieri
Department of Experimental and Clinical Medicine, University of Florence, Italy
Costanza Bogani
Department of Experimental and Clinical Medicine, University of Florence, Italy
Denis Clay
INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France;INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France
Frederic Torossian
INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France;INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France
Jean-Jacques Lataillade
INSERM UMR-S1197, Paul Brousse Hospital, Paris-Sud University, Villejuif, France;Department of Experimental and Clinical Medicine, University of Florence, Italy
Hans C. Hasselbach
Department of Hematology, Herlev University Hospital, Copenhagen, Denmark
Heinz Gisslinger
Department of Hematology, University Klinik Fur Innere Medizin, Vienna, Austria
Jean-Loup Demory
Université Catholique de Lille, France;French Intergroup on Myeloproliferative Neoplasms (FIM), France
Brigitte Dupriez
French Intergroup on Myeloproliferative Neoplasms (FIM), France;Department of Hematology, Dr Schaffner Hospital, Lens, France
Claude Boucheix
INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France;Inserm U935, Paul Brousse Hospital, Paris-Sud University, Villejuif, France
Eric Rubinstein
INSERM UMS33, Paul Brousse Hospital, Paris-Sud University, Villejuif, France;Inserm U935, Paul Brousse Hospital, Paris-Sud University, Villejuif, France
Sophie Amsellem
Department of Hematology, Gustave Roussy Institute, Villejuif, France
Alessandro M. Vannucchi
Department of Experimental and Clinical Medicine, University of Florence, Italy
Marie-Caroline Le Bousse-Kerdilès
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
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.