BACKGROUND: Demonstrating the presence of myelodysplastic syndrome (MDS)-specific molecular abnormalities can aid in diagnosis and patient management. We explored the potential of using peripheral blood (PB) cell-free DNA (cf-DNA) and next-generation sequencing (NGS). MATERIALS AND METHODS: We performed NGS on a panel of 14 target genes using total nucleic acid extracted from the plasma of 16 patients, all of whom had confirmed diagnoses for early MDS with blasts <5%. PB cellular DNA from the same patients was sequenced using conventional Sanger sequencing and NGS. RESULTS: Deep sequencing of the cf-DNA identified one or more mutated gene(s), confirming the diagnosis of MDS in all cases. Five samples (31%) showed abnormalities in cf-DNA by NGS that were not detected by Sanger sequencing on cellular PB DNA. NGS of PB cell DNA showed the same findings as those of cf-DNA in four of five patients, but failed to show a mutation in the RUNX1 gene that was detected in one patient's cf-DNA. Mutant allele frequency was significantly higher in cf-DNA compared with cellular DNA (p = 0.008). CONCLUSION: These data suggest that cf-DNA when analyzed using NGS is a reliable approach for detecting molecular abnormalities in MDS and should be used to determine if bone marrow aspiration and biopsy are necessary.
BACKGROUND: Demonstrating the presence of myelodysplastic syndrome (MDS)-specific molecular abnormalities can aid in diagnosis and patient management. We explored the potential of using peripheral blood (PB) cell-free DNA (cf-DNA) and next-generation sequencing (NGS). MATERIALS AND METHODS: We performed NGS on a panel of 14 target genes using total nucleic acid extracted from the plasma of 16 patients, all of whom had confirmed diagnoses for early MDS with blasts <5%. PB cellular DNA from the same patients was sequenced using conventional Sanger sequencing and NGS. RESULTS: Deep sequencing of the cf-DNA identified one or more mutated gene(s), confirming the diagnosis of MDS in all cases. Five samples (31%) showed abnormalities in cf-DNA by NGS that were not detected by Sanger sequencing on cellular PB DNA. NGS of PB cell DNA showed the same findings as those of cf-DNA in four of five patients, but failed to show a mutation in the RUNX1 gene that was detected in one patient's cf-DNA. Mutant allele frequency was significantly higher in cf-DNA compared with cellular DNA (p = 0.008). CONCLUSION: These data suggest that cf-DNA when analyzed using NGS is a reliable approach for detecting molecular abnormalities in MDS and should be used to determine if bone marrow aspiration and biopsy are necessary.
Authors: Caitlin M Stewart; Prachi D Kothari; Florent Mouliere; Richard Mair; Saira Somnay; Ryma Benayed; Ahmet Zehir; Britta Weigelt; Sarah-Jane Dawson; Maria E Arcila; Michael F Berger; Dana Wy Tsui Journal: J Pathol Date: 2018-03-12 Impact factor: 7.996
Authors: Peter Valent; Attilio Orazi; David P Steensma; Benjamin L Ebert; Detlef Haase; Luca Malcovati; Arjan A van de Loosdrecht; Torsten Haferlach; Theresia M Westers; Denise A Wells; Aristoteles Giagounidis; Michael Loken; Alberto Orfao; Michael Lübbert; Arnold Ganser; Wolf-Karsten Hofmann; Kiyoyuki Ogata; Julie Schanz; Marie C Béné; Gregor Hoermann; Wolfgang R Sperr; Karl Sotlar; Peter Bettelheim; Reinhard Stauder; Michael Pfeilstöcker; Hans-Peter Horny; Ulrich Germing; Peter Greenberg; John M Bennett Journal: Oncotarget Date: 2017-07-05