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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Residual Neoplasm
Acute Myeloid Leukemia
Mutation
DNA
fms-Like Tyrosine Kinase 3
Bone Marrow
Randomized Controlled Trials
Polymerase Chain Reaction
Survival

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A next-generation sequencing-based assay for minimal residual disease assessment in AML patients with FLT3-ITD mutations. / Levis, Mark J; Perl, Alexander E.; Altman, Jessica K.; Gocke, Christopher; Bahceci, Erkut; Hill, Jason; Liu, Chaofeng; Xie, Zhiyi; Carson, Andrew R.; McClain, Valerie; Stenzel, Timothy T.; Miller, Jeffrey E.

In: Blood advances, Vol. 2, No. 8, 24.04.2018, p. 825-831.

Research output: Contribution to journalArticle

Levis, MJ, Perl, AE, Altman, JK, Gocke, C, Bahceci, E, Hill, J, Liu, C, Xie, Z, Carson, AR, McClain, V, Stenzel, TT & Miller, JE 2018, 'A next-generation sequencing-based assay for minimal residual disease assessment in AML patients with FLT3-ITD mutations', Blood advances, vol. 2, no. 8, pp. 825-831. https://doi.org/10.1182/bloodadvances.2018015925
Levis, Mark J ; Perl, Alexander E. ; Altman, Jessica K. ; Gocke, Christopher ; Bahceci, Erkut ; Hill, Jason ; Liu, Chaofeng ; Xie, Zhiyi ; Carson, Andrew R. ; McClain, Valerie ; Stenzel, Timothy T. ; Miller, Jeffrey E. / A next-generation sequencing-based assay for minimal residual disease assessment in AML patients with FLT3-ITD mutations. In: Blood advances. 2018 ; Vol. 2, No. 8. pp. 825-831.
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