title: Routine clinical mutation profiling using next generation sequencing and a customized gene panel improves diagnostic precision in myeloid neoplasms.
abstract: Microscopic examination of myelodysplastic syndromes (MDS) and myelodysplastic-myeloproliferative neoplasms (MDS/MPN) may be challenging because morphological features can overlap with those of reactive states. Demonstration of clonal hematopoiesis provides a diagnostic clue and has become possible by comprehensive mutation profiling of a number of frequently mutated genes, some of them with large coding regions.To emphasize the potential benefit of NGS in hematopathology we present sequencing results from routinely processed formalin-fixed and paraffin-embedded (FFPE) bone marrow trephines (n = 192). A customized amplicon-based gene panel including 23 genes frequently mutated in myeloid neoplasms was established and implemented. Thereby, 629,691 reads per sample (range 179,847-1,460,412) and a mean coverage of 2,702 (range 707-6,327) could be obtained, which are sufficient for comprehensive mutational profiling. Seven samples failed in sequencing (3.6%). In 185 samples we found in total 269 pathogenic variants (mean 1.4 variants per patient, range 0-5), 125 Patients exhibit at least one pathogenic mutation (67.6%). Variants show allele frequencies ranging from 6.7% up to 95.7%. Most frequently mutated genes were TET2 (28.7%), SRSF2 (19.5%), ASXL1 (8.6%) and U2AF1 (8.1%). The mutation profiling increases the diagnostic precision and adds prognostic information.
authors.affiliation: Institute of Pathology, Medizinische Hochschule Hannover, Hannover, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Bartels
authors.affiliation: Institute of Pathology, Medizinische Hochschule Hannover, Hannover, Germany.
authors.forename: Elisa
authors.initials: E
authors.lastname: Schipper
authors.affiliation: Institute of Pathology, Medizinische Hochschule Hannover, Hannover, Germany.
authors.forename: Britta
authors.initials: B
authors.lastname: Hasemeier
authors.affiliation: Institute of Pathology, Medizinische Hochschule Hannover, Hannover, Germany.
authors.forename: Hans
authors.initials: H
authors.lastname: Kreipe
authors.affiliation: Institute of Pathology, Medizinische Hochschule Hannover, Hannover, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Lehmann
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000235:genetics; D001706:Biopsy; D001853:Bone Marrow / Q000473:pathology; D004252:DNA Mutational Analysis / Q000379:methods; D004268:DNA-Binding Proteins / Q000235:genetics; D049308:Dioxygenases; D005787:Gene Frequency; D019337:Hematologic Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008875:Middle Aged; D009154:Mutation; D009190:Myelodysplastic Syndromes / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D054437:Myelodysplastic-Myeloproliferative Diseases / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D011379:Prognosis; D011518:Proto-Oncogene Proteins / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D017422:Sequence Analysis, DNA; D000068103:Serine-Arginine Splicing Factors / Q000235:genetics; D000072668:Splicing Factor U2AF / Q000235:genetics; D055815:Young Adult
chemical_list: C473949:ASXL1 protein, human; D014408:Biomarkers, Tumor; D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D012097:Repressor Proteins; D000072668:Splicing Factor U2AF; C076753:U2AF1 protein, human; C073288:SRSF2 protein, human; D000068103:Serine-Arginine Splicing Factors; D049308:Dioxygenases; C541081:TET2 protein, human
keywords: FFPE; MDS; Pathology Section; hematopathology; myeloid neoplasms; next-generation sequencing