title: The detection of SRSF2 mutations in routinely processed bone marrow biopsies is useful in the diagnosis of chronic myelomonocytic leukemia.
abstract: Diagnosis of chronic myelomonocytic leukemia (CMML) is based on a combination of clinical, laboratory, and morphological parameters, including persistent peripheral blood monocytosis. Recently, mutations of serine/arginine-rich splicing factor 2 (SRSF2) have been identified in 40% to 50% of CMMLs and occasionally in other myeloid disorders. In this study, we established a robust assay for the detection of SRSF2 mutations in decalcified, paraffin-embedded bone marrow (BM) biopsies and investigated its diagnostic utility. BM biopsies of 78 patients with myeloid neoplasms, including 36 CMMLs, 22 myelodysplastic syndromes (MDS), and 20 Ph- myeloproliferative neoplasms (MPN) were analyzed. The region around hot spot P95 in exon 1 of SRSF2 was amplified and bidirectionally sequenced. In addition, a restriction fragment length polymorphism analysis was established. The JAK2 V617F mutation was investigated by allele-specific polymerase chain reaction. SRSF2 mutations were identified in 16 (44%) of 36 CMMLs, including 1 of 3 cases with associated systemic mastocytosis, 4 (20%) of 20 Ph- MPN, and 1 (4.5%) of 22 MDS. Restriction fragment length polymorphism analysis detected all mutations with the exception of a single P95A. Of note, 2 cases of JAK2 V617F+ primary myelofibrosis with SRSF2 mutation initially were diagnosed as CMML based on significant peripheral blood monocytosis. In CMML, no correlation with histopathology and/or clinical parameters was observed, but SRSF2 mutations were associated with normal karyotype (P < .001). In summary, SRSF2 mutations are frequent in CMML and a useful diagnostic feature demonstrable in BM biopsies, allowing a definitive diagnosis for cases with minimal dysplasia and normal karyotype. The role of SRSF2 mutations in cases with hybrid features between primary myelofibrosis and CMML needs further investigation.
authors.affiliation: Institute of Pathology and Neuropathology Comprehensive Cancer Center Tuebingen and University of Tuebingen, 72076 Tuebingen, Germany.
authors.forename: Birgit
authors.initials: B
authors.lastname: Federmann
authors.affiliation: Institute of Pathology and Neuropathology Comprehensive Cancer Center Tuebingen and University of Tuebingen, 72076 Tuebingen, Germany.
authors.forename: Manuel
authors.initials: M
authors.lastname: Abele
authors.affiliation: Institute of Pathology and Neuropathology Comprehensive Cancer Center Tuebingen and University of Tuebingen, 72076 Tuebingen, Germany; Department of Pathology, Hospital Universitario Miguel Servet, 50001 Zaragoza, Spain.
authors.forename: David Santiago
authors.initials: DS
authors.lastname: Rosero Cuesta
authors.affiliation: Department of Internal Medicine, Hematology and Oncology, Comprehensive Cancer Center Tuebingen and University of Tuebingen, 72076 Tuebingen, Germany.
authors.forename: Wichard
authors.initials: W
authors.lastname: Vogel
authors.affiliation: Department of Molecular and Translational Medicine, Pathology Section, University of Brescia, 25100 Brescia, Italy.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Boiocchi
authors.affiliation: Department of Pathology, Hospital Universitario Miguel Servet, 50001 Zaragoza, Spain.
authors.forename: Lothar
authors.initials: L
authors.lastname: Kanz
authors.affiliation: Institute of Pathology and Neuropathology Comprehensive Cancer Center Tuebingen and University of Tuebingen, 72076 Tuebingen, Germany.
authors.forename: Leticia
authors.initials: L
authors.lastname: Quintanilla-Martinez
authors.affiliation: Department of Pathology and Laboratory Medicine, Weill Cornell Medical College/New York Presbyterian Hospital, 10065 NY, USA.
authors.forename: Attilio
authors.initials: A
authors.lastname: Orazi
authors.affiliation: Institute of Pathology and Neuropathology Comprehensive Cancer Center Tuebingen and University of Tuebingen, 72076 Tuebingen, Germany.
authors.forename: Irina
authors.initials: I
authors.lastname: Bonzheim
authors.affiliation: Institute of Pathology and Neuropathology Comprehensive Cancer Center Tuebingen and University of Tuebingen, 72076 Tuebingen, Germany. Electronic address: falko.fend@med.uni-tuebingen.de.
authors.forename: Falko
authors.initials: F
authors.lastname: Fend
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001706:Biopsy; D001853:Bone Marrow / Q000473:pathology*; D001856:Bone Marrow Examination; D005260:Female; D006801:Humans; D015477:Leukemia, Myelomonocytic, Chronic / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics*; D012261:Ribonucleoproteins / Q000235:genetics*; D000068103:Serine-Arginine Splicing Factors
chemical_list: D009687:Nuclear Proteins; D012261:Ribonucleoproteins; C073288:SRSF2 protein, human; D000068103:Serine-Arginine Splicing Factors
keywords: Bone marrow biopsy; Chronic myelomonocytic leukemia; JAK2 mutation; Myeloproliferative neoplasms; SRSF2 mutation