title: Clinical relevance of sensitive and quantitative STAT3 mutation analysis using next-generation sequencing in T-cell large granular lymphocytic leukemia.
abstract: Diagnosis of T-cell large granular lymphocytic leukemia (T-LGL) is often challenging because clinical and laboratory characteristics are overlapping with nonneoplastic conditions. Recently, mutation in the STAT3 gene has been identified as a recurrent genetic abnormality in T-LGL. STAT3 mutation, therefore, represents a promising marker in T-LGL diagnostics. We developed a new quantitative next-generation sequencing assay that allows sensitive analysis of the STAT3 gene. The assay was used to study the utility of STAT3 mutation analysis as a diagnostic tool in T-LGL. The study included 16 T-LGL patients. A total of 15 mutations, including 2 new mutations (G618R and K658R), were detected in 12 patients (75%), with mutation levels ranging from 2.5% to 45.6% mutation-positive alleles. Next-generation sequencing detected 50% more mutations than Sanger sequencing. Blood samples from 20 healthy blood donors all tested negative, thus demonstrating the specificity of the assay. The results also indicated that mutation levels in blood and bone marrow are not systematically different, and next-generation sequencing-based STAT3 mutation analysis represents a sensitive method for monitoring residual disease as demonstrated in a patient receiving pentostatin. We demonstrate the clinical relevance of next-generation sequencing-based STAT3 mutation analysis, which represents a sensitive and specific diagnostic marker in T-LGL that allows assessment of molecular residual disease, which may improve clinical decision making.
authors.affiliation: Department of Pathology, Odense University Hospital, Odense, Denmark. Electronic address: thomas.kielsgaard.kristensen@rsyd.dk.
authors.forename: Thomas
authors.initials: T
authors.lastname: Kristensen
authors.affiliation: Department of Clinical Genetics, Odense University Hospital, Odense, Denmark; Division of Human Genetics, Clinical Institute, University of Southern Denmark, Odense, Denmark.
authors.forename: Martin
authors.initials: M
authors.lastname: Larsen
authors.affiliation: Department of Hematology, Odense University Hospital, Odense, Denmark.
authors.forename: Annika
authors.initials: A
authors.lastname: Rewes
authors.affiliation: Department of Hematology, Odense University Hospital, Odense, Denmark.
authors.forename: Henrik
authors.initials: H
authors.lastname: Frederiksen
authors.affiliation: Department of Clinical Genetics, Odense University Hospital, Odense, Denmark; Division of Human Genetics, Clinical Institute, University of Southern Denmark, Odense, Denmark.
authors.forename: Mads
authors.initials: M
authors.lastname: Thomassen
authors.affiliation: Department of Pathology, Odense University Hospital, Odense, Denmark.
authors.forename: Michael Boe
authors.initials: MB
authors.lastname: Møller
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D004252:DNA Mutational Analysis; D005260:Female; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D054066:Leukemia, Large Granular Lymphocytic / Q000175:diagnosis / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D015203:Reproducibility of Results; D050796:STAT3 Transcription Factor / Q000235:genetics*; D012680:Sensitivity and Specificity
chemical_list: D050796:STAT3 Transcription Factor