Abstract
JAK2(V617F) is the predominant mutation in myeloproliferative neoplasms (MPN). Modeling MPN in a human context might be helpful for the screening of molecules targeting JAK2 and its intracellular signaling. We describe here the derivation of induced pluripotent stem (iPS) cell lines from 2 polycythemia vera patients carrying a heterozygous and a homozygous mutated JAK2(V617F), respectively. In the patient with homozygous JAK2(V617F), additional ASXL1 mutation and chromosome 20 allowed partial delineation of the clonal architecture and assignation of the cellular origin of the derived iPS cell lines. The marked difference in the response to erythropoietin (EPO) between homozygous and heterozygous cell lines correlated with the constitutive activation level of signaling pathways. Strikingly, heterozygous iPS cells showed thrombopoietin (TPO)-independent formation of megakaryocytic colonies, but not EPO-independent erythroid colony formation. JAK2, PI3K and HSP90 inhibitors were able to block spontaneous and EPO-induced growth of erythroid colonies from GPA(+)CD41(+) cells derived from iPS cells. Altogether, this study brings the proof of concept that iPS can be used for studying MPN pathogenesis, clonal architecture, and drug efficacy.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Cells, Cultured
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Erythropoietin / pharmacology
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Humans
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Induced Pluripotent Stem Cells / cytology*
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Induced Pluripotent Stem Cells / metabolism*
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Janus Kinase 2 / genetics
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Janus Kinase 2 / metabolism*
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Myeloproliferative Disorders / genetics
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Myeloproliferative Disorders / metabolism*
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Phosphatidylinositol 3-Kinases / genetics
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Phosphatidylinositol 3-Kinases / metabolism
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Repressor Proteins / genetics
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Repressor Proteins / metabolism
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Signal Transduction / drug effects
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Signal Transduction / genetics
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Thrombopoietin / pharmacology
Substances
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ASXL1 protein, human
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Repressor Proteins
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Erythropoietin
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Thrombopoietin
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Phosphatidylinositol 3-Kinases
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Janus Kinase 2
Grants and funding
This work was supported by grants from Agence Nationale de la Recherche (ANR) (ANR-blanc Megon 2009 and thrombocytosis 2011; ANR-Blanc 2010 Epigenome) and from the Association pour la Recherche sur le Cancer (ARC) (ontogenèse et mégacaryopoïèse normale et pathologique, and ARC libre 2012). Labex GR-Ex (IP, VW) is funded by the program "Investissements d'avenir". TL was supported by a fellowship from ANR-blanc Megon. GL was supported by a postdoctoral fellowship from the Ile de France Cancéropôle and Institut National du Cancer (INCA). LS and JS were supported by PhD grants from the Ile de France region (Cancéropôle, Département d'information médicale (DIM) cellule souche, Fondation pour la Recherche Médicale (FRM)). ND, HR and WV are recipients of a research fellowship from Centre Hospitalier Universitaire (CHU Bordeaux)-INSERM, Assistance Publique-Hopitaux de Paris (AP-HP)-INSERM and Institut Gustave Roussy (IGR)-INSERM (contrats d'interface). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.