Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Daga, S; Rosenberger, A; Kashofer, K; Heitzer, E; Quehenberger, F; Halbwedl, I; Graf, R; Krisper, N; Prietl, B; Höfler, G; Reinisch, A; Zebisch, A; Sill, H; Wölfler, A.
Sensitive and broadly applicable residual disease detection in acute myeloid leukemia using flow cytometry-based leukemic cell enrichment followed by mutational profiling.
Am J Hematol. 2020; 95(10):1148-57
Doi: 10.1002/ajh.25918
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- Führende Autor*innen der Med Uni Graz
- Daga Shruti
- Wölfler Albert
- Co-Autor*innen der Med Uni Graz
- Graf Ricarda
- Halbwedl Iris
- Heitzer Ellen
- Höfler Gerald
- Kashofer Karl
- Krisper Nina
- Prietl Barbara
- Quehenberger Franz
- Reinisch Andreas
- Schlacher Angelika
- Sill Heinz
- Zebisch Armin
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- Abstract:
- Persistent measurable residual disease (MRD) is an increasingly important prognostic marker in acute myeloid leukemia (AML). Currently, MRD is determined by multi-parameter flow cytometry (MFC) or PCR-based methods detecting leukemia-specific fusion transcripts and mutations. However, while MFC is highly operator-dependent and difficult to standardize, PCR-based methods are only available for a minority of AML patients. Here we describe a novel, highly sensitive and broadly applicable method for MRD detection by combining MFC-based leukemic cell enrichment using an optimized combinatorial antibody panel targeting CLL-1, TIM-3, CD123 and CD117, followed by mutational analysis of recurrently mutated genes in AML. In dilution experiments this method showed a sensitivity of 10-4 to 10-5 for residual disease detection. In prospectively collected remission samples this marker combination allowed for a median 67-fold cell enrichment with sufficient DNA quality for mutational analysis using next generation sequencing (NGS) or digital PCR in 39 out of 41 patients. Twenty-one samples (53.8%) tested MRD positive, whereas 18 (46.2%) were negative. With a median follow-up of 559 days, 71.4% of MRD positive (15/21) and 27.8% (5/18) of MRD negative patients relapsed (P = .007). The cumulative incidence of relapse (CIR) was higher for MRD positive patients (5-year CIR: 90.5% vs 28%, P < .001). In multivariate analysis, MRD positivity was a prominent factor for CIR. Thus, MFC-based leukemic cell enrichment using antibodies against CLL-1, TIM-3, CD123 and CD117 followed by mutational analysis allows high sensitive MRD detection and is informative on relapse risk in the majority of AML patients.