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

Mark J Levis1, Alexander E Perl2, Jessica K Altman3

  • 1Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD.

Blood Advances
|April 13, 2018
PubMed

Insights

A new, highly sensitive next-generation sequencing assay accurately detects minimal residual disease (MRD) in fms-like tyrosine kinase 3 (FLT3-ITD) mutated acute myeloid leukemia (AML), improving upon existing methods for guiding therapy.

Area of Science:

  • Hematology
  • Molecular Diagnostics
  • Oncology

Background:

  • Internal tandem duplications in fms-like tyrosine kinase 3 (FLT3-ITD) are prevalent in acute myeloid leukemia (AML) and associated with poor prognosis.
  • Accurate detection of minimal residual disease (MRD) in FLT3-ITD AML is crucial for guiding treatment decisions.
  • Current MRD assays for FLT3-ITD AML lack sufficient sensitivity.

Purpose of the Study:

  • To develop and validate a sensitive and specific next-generation sequencing (NGS)-based assay for MRD detection in FLT3-ITD mutated AML.
  • To compare the sensitivity of the novel assay with existing methods.
  • To evaluate the assay's utility in assessing mutation burden and its correlation with overall survival in relapsed/refractory AML patients.

Main Methods:

  • Development of a novel NGS-based assay for FLT3-ITD mutation detection.
  • Validation using spiked DNA samples to determine sensitivity (≥1 FLT3-ITD cell in 10,000).
  • Assay validation in bone marrow samples from patients with FLT3-ITD AML in remission and relapsed/refractory AML.

Main Results:

  • The developed NGS assay demonstrated high sensitivity and specificity for FLT3-ITD MRD detection.
  • The assay is two orders of magnitude more sensitive than current PCR or NGS-based assays.
  • A correlation was observed between mutation burden, as detected by the assay, and overall survival in relapsed/refractory AML patients treated with gilteritinib.

Conclusions:

  • The novel NGS-based MRD assay offers significantly improved sensitivity and specificity for FLT3-ITD AML.
  • This assay has the potential to refine MRD monitoring and guide therapeutic strategies in FLT3-ITD AML.
  • Prospective validation in ongoing clinical trials will further establish the assay's clinical utility.

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