title: Cell autonomous expression of CXCL-10 in JAK2V617F-mutated MPN.
abstract: OBJECTIVE
  Myeloproliferative neoplasms (MPN) are clonal disorders of hematopoietic stem- and progenitor cells. Mutation of Janus-Kinase 2 (JAK2) is the most frequent genetic event detected in Philadelphia-negative MPN. In advanced phases, the clinical hallmark of the disease is a striking inflammatory syndrome. So far, the cellular and molecular basis of inflammation is not fully understood. We, therefore, sought to investigate the relationship of activating JAK2 mutation and aberrant cytokine expression in MPN.
  
  
  METHODS
  Cytokine array was performed to identify Jak2V617F-related cytokine expression and secretion. CXCL10 mRNA expression was analyzed by qPCR in peripheral blood cells. To exclude paracrine/autocrine stimulation as a potential mechanism, we generated Ba/F3-EpoR-JAK2WT or EpoR-JAK2V617F cells lacking CXCL10 receptor. Pharmacologic inhibition of JAK2 kinase was achieved by JAK-Inhibitor treatment. Signaling pathways and downstream effectors were characterized by Western blotting, immunofluorescence microscopy, luciferase reporter assays, qPCR, and chromatin-immunoprecipitation studies.
  
  
  RESULTS
  We identified CXCL10 as the most highly induced cytokine in JAK2-mutated cell lines. In MPN patients, CXCL10 is highly expressed in JAK2V617F but not JAK2WT MPN or healthy donor controls. Moreover, CXCL10 expression correlates with JAK2V617F allelic burden. High CXCL10 correlates with the presence of clinical risk factors but not with clinical symptoms and quality of life. Pharmacologic inhibition of mutated JAK2 kinase inhibits CXCL10 expression. NFκB signaling is activated downstream of JAK2V617F receptor and directly induces CXCL10 expression.
  
  
  CONCLUSIONS
  Our data provide first evidence for a link between oncogenic JAK2V617F signaling and cell intrinsic induction of CXCL10 induced by activated NFkB signaling.
authors.affiliation: Innere Medizin II, Hämatologie und Onkologie, Universitätsklinikum Jena, Am Klinikum 1, 07747, Jena, Germany.|Leibniz Institute on Aging, Fritz-Lipmann-Institute, Jena, Germany.
authors.forename: Tina M
authors.initials: TM
authors.lastname: Schnöder
authors.affiliation: Department of Hematology and Oncology, Otto-von-Guericke University Medical Center, Magdeburg, Germany.
authors.forename: Judith
authors.initials: J
authors.lastname: Eberhardt
authors.affiliation: Department of Hematology and Oncology, Otto-von-Guericke University Medical Center, Magdeburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Koehler
authors.affiliation: Leibniz Institute on Aging, Fritz-Lipmann-Institute, Jena, Germany.|Department of Genetics, Friedrich-Schiller-University, Jena, Germany.
authors.forename: Holger B
authors.initials: HB
authors.lastname: Bierhoff
authors.affiliation: Department of Cardiology, Otto-von-Guericke University Medical Center, Magdeburg, Germany.
authors.forename: Sönke
authors.initials: S
authors.lastname: Weinert
authors.affiliation: Department of Hematology and Oncology, Otto-von-Guericke University Medical Center, Magdeburg, Germany.
authors.forename: Akhilesh Datt
authors.initials: AD
authors.lastname: Pandey
authors.affiliation: Department of Hematology and Oncology, Otto-von-Guericke University Medical Center, Magdeburg, Germany.
authors.forename: Subbaiah Chary
authors.initials: SC
authors.lastname: Nimmagadda
authors.affiliation: Department of Hematology and Oncology, Otto-von-Guericke University Medical Center, Magdeburg, Germany.
authors.forename: Denise
authors.initials: D
authors.lastname: Wolleschak
authors.affiliation: Institut für Pathologie, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Korinna
authors.initials: K
authors.lastname: Jöhrens
authors.affiliation: Department of Hematology and Oncology, Otto-von-Guericke University Medical Center, Magdeburg, Germany.|Collaborative Research Cluster 854 (CRC854), Medical Faculty, University Hospital Magdeburg, Magdeburg, Germany.
authors.forename: Thomas
authors.identifier: http://orcid.org/0000-0003-2438-1955
authors.initials: T
authors.lastname: Fischer
authors.affiliation: Innere Medizin II, Hämatologie und Onkologie, Universitätsklinikum Jena, Am Klinikum 1, 07747, Jena, Germany. Florian.Heidel@med.uni-jena.de.|Leibniz Institute on Aging, Fritz-Lipmann-Institute, Jena, Germany. Florian.Heidel@med.uni-jena.de.|Collaborative Research Cluster 854 (CRC854), Medical Faculty, University Hospital Magdeburg, Magdeburg, Germany. Florian.Heidel@med.uni-jena.de.
authors.forename: Florian H
authors.identifier: http://orcid.org/0000-0003-2438-1955
authors.initials: FH
authors.lastname: Heidel
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D054357:Chemokine CXCL10 / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D009154:Mutation; D009196:Myeloproliferative Disorders / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics
chemical_list: C114753:CXCL10 protein, human; D054357:Chemokine CXCL10; C485433:Cxcl10 protein, mouse; D012333:RNA, Messenger; C507924:JAK2 protein, human; D053614:Janus Kinase 2
keywords: CXCL10; ET; IP10; Inflammation; MF; MPN; PV