title: Detection of the single hotspot mutation in the JH2 pseudokinase domain of Janus kinase 2 in bone marrow trephine biopsies derived from chronic myeloproliferative disorders.
abstract: The recent discovery of a single point mutation in the JH2 pseudokinase domain of Janus kinase 2 (JAK2) in a considerable fraction of patients has shed light on the molecular pathomechanism in Philadelphia chromosome-negative chronic myeloproliferative disorders (Ph- CMPDs). We established a robust and reliable method for detection of the JAK2 mutation in bone marrow cells derived from archival bone marrow trephines based on polymerase chain reaction and subsequent restriction site analysis. In a series of proven Ph- CMPDs classified according to World Health Organization criteria (n = 79), we detected the JAK2 mutation in 90% of polycythemia vera, 22% of cellular prefibrotic chronic idiopathic myelofibrosis, 60% of advanced chronic idiopathic myelofibrosis, and 27% of essential thrombocythemia. JAK2 mutation was not detected in Ph+ chronic myeloid leukemia (n = 5), acute myeloid leukemia (n = 10), acute lymphoblastic leukemia (n = 10), secondary erythrocytosis (n = 10), or normal bone marrow (n = 10). Restriction site analysis was also suitable for unfixed cell populations derived from peripheral blood and bone marrow aspirates. Besides providing support in the differential diagnosis of reactive versus neoplastic myeloproliferations, this newly developed assay reveals considerable overlaps between histologically different disease entities, indicating that additional genetic alterations might be responsible for the established differences of CMPD subentities.
authors.affiliation: Institute of Pathology, Medizinische Hochschule Hannover, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany. bock.oliver@mh-hannover.de
authors.forename: Oliver
authors.initials: O
authors.lastname: Bock
authors.forename: Guntram
authors.initials: G
authors.lastname: Büsche
authors.forename: Christina
authors.initials: C
authors.lastname: Koop
authors.forename: Sabine
authors.initials: S
authors.lastname: Schröter
authors.forename: Thomas
authors.initials: T
authors.lastname: Buhr
authors.forename: Hans
authors.initials: H
authors.lastname: Kreipe
mesh_terms: D001483:Base Sequence; D001706:Biopsy; D001853:Bone Marrow / Q000378:metabolism*; D002908:Chronic Disease; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D006410:Hematopoiesis; D006801:Humans; D053614:Janus Kinase 2; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009196:Myeloproliferative Disorders / Q000201:enzymology* / Q000235:genetics* / Q000473:pathology; D009369:Neoplasms; D011505:Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D015183:Restriction Mapping
chemical_list: D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D011505:Protein-Tyrosine Kinases; D016044:Fusion Proteins, bcr-abl; C507924:JAK2 protein, human; D053614:Janus Kinase 2