A next-generation sequencing-based assay for minimal residual disease assessment in AML patients with FLT3-ITD mutations

Mark J Levis, Alexander E. Perl, Jessica K. Altman, Christopher Gocke, Erkut Bahceci, Jason Hill, Chaofeng Liu, Zhiyi Xie, Andrew R. Carson, Valerie McClain, Timothy T. Stenzel, Jeffrey E. Miller

Research output: Contribution to journalArticle

Abstract

Internal tandem duplications in fms-like tyrosine kinase 3 (FLT3-ITDs) are common in acute myeloid leukemia (AML) and confer a poor prognosis. A sensitive and specific assay for the detection of minimal residual disease (MRD) in FLT3-ITD mutated AML could guide therapy decisions. Existing assays for MRD in FLT3-ITD AML have not been particularly useful because of limited sensitivity. We developed a sensitive and specific MRD assay for FLT3-ITD mutations using next-generation sequencing. The initial validation of this assay was performed by spiking fixed amounts of mutant DNA into wild-type DNA to establish a sensitivity of detection equivalent to ≥1 FLT3-ITD-containing cell in 10 000, with a minimum input of 100 000 cell equivalents of DNA. We subsequently validated the assay in bone marrow samples from patients with FLT3-ITD AML in remission. Finally, we analyzed bone marrow samples from 80 patients with FLT3-ITD relapsed/refractory AML participating in a trial of a novel FLT3 inhibitor, gilteritinib, and demonstrated a relationship between the mutation burden, as detected by the assay, and overall survival. This novel MRD assay is specific and 2 orders of magnitude more sensitive than currently available polymerase chain reaction- or next-generation sequencing-based FLT3-ITD assays. The assay is being prospectively validated in ongoing randomized clinical trials.

Original languageEnglish (US)
Pages (from-to)825-831
Number of pages7
JournalBlood advances
Volume2
Issue number8
DOIs
StatePublished - Apr 24 2018

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  • Cite this

    Levis, M. J., Perl, A. E., Altman, J. K., Gocke, C., Bahceci, E., Hill, J., Liu, C., Xie, Z., Carson, A. R., McClain, V., Stenzel, T. T., & Miller, J. E. (2018). A next-generation sequencing-based assay for minimal residual disease assessment in AML patients with FLT3-ITD mutations. Blood advances, 2(8), 825-831. https://doi.org/10.1182/bloodadvances.2018015925