Abstract
BackgroundThe gain-of-function mutation JAK2V617F is frequently found in Philadelphia-chromosome-negative myeloproliferative neoplasm (MPN) patients. However, the tumorigenic properties of JAK2V617F have mostly been characterized in in vivo and in vitro murine models due to the lack of appropriate human cell lines.MethodsUsing the multipotent hematologic cell line UT-7/GM, we established D9, a novel human cell line that expresses JAK2V617F upon tetracycline addition. We assessed cellular differentiation in UT-7/GM cells when JAK2V617F was induced, and we used microarrays to analyze changes in mRNA expression caused by JAK2V617F.ResultsUsing the human D9 cell line, we demonstrated that the induction of JAK2V617F leads to cytokine-independent cell growth with increased STAT activation and erythroid differentiation, mimicking the characteristics observed in polycythemia vera, making it a suitable in vitro model for studying this disorder. Interestingly, JAK2V617F-dependent erythroid cell differentiation was blocked when GM-CSF was added to the culture, suggesting that the GM-CSF pathway antagonizes JAK2V617F-induced erythroid cell differentiation. Our microarray analysis identified several genes involved in inflammasome activation, such as AIM2, IL1B, and CASP1, which were significantly up-regulated in JAK2V617F-induced cells.ConclusionsThe observed inflammasome activation following JAK2V617F induction is consistent with a recent report demonstrating the involvement of IL1B in myelofibrosis development in a JAK2V617F model mouse. These results indicate that the D9 cell line should be useful for characterizing the signaling pathways downstream of JAK2V617F, allowing for the identification of effector molecules that contribute to the development of MPN.Electronic supplementary materialThe online version of this article (doi:10.1186/s40164-016-0032-7) contains supplementary material, which is available to authorized users.
Highlights
The gain-of-function mutation JAK2V617F is frequently found in Philadelphia-chromosome-negative myeloproliferative neoplasm (MPN) patients
Unlike chronic neutrophilic leukemia (CNL) that is associated with CSF3R mutation [1,2,3], polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF) share a common acquired mutation, JAK2V617F, which is found in approximately 95 % of PV patients and 50 % of ET and PMF patients [4,5,6,7]
These cells will be an invaluable tool with which to investigate the downstream effectors of JAK2V617F, which should allow us to elucidate the relationship between MPNs and the JAK2V617F mutation
Summary
The gain-of-function mutation JAK2V617F is frequently found in Philadelphia-chromosome-negative myeloproliferative neoplasm (MPN) patients. JAK2V617F activation stimulates several downstream signaling networks, including the STAT3, STAT5, ERK/MAP kinase, and PI3 kinase/Akt pathways, leading to cytokine-independent cell growth (reviewed in [10]) Activation of these pathways can partially explain the phenotypes observed in MPN patient. We used UT-7/GM as the basis for the cell line, which has the unique ability to differentiate into erythrocytes or megakaryocytes, depending on the presence of EPO or thrombopoietin (TPO), respectively [14, 15] These cells will be an invaluable tool with which to investigate the downstream effectors of JAK2V617F, which should allow us to elucidate the relationship between MPNs and the JAK2V617F mutation
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