Prospective feasibility of a minimal BH3 profiling assay in acute myeloid leukemia
Kim Pacchiardi1,2, Victoire de Marcellus1, Tony Huynh3
1Université Paris Cité, Génomes, biologie cellulaire et thérapeutique U944, INSERM, CNRS, Paris, France.
Abstract:
BH3 profiling can assess global mitochondrial priming and dependence of leukemic cells on specific BH3 anti-apoptotic proteins such as BCL-2. In acute myeloid leukemia (AML), proof-of-concept prognostic studies have been performed on archived samples variably accounting for molecular genetics. We undertook a single-center feasibility study of a simplified flow-based assay to determine the absolute mitochondrial priming and BCL-2 dependence in consecutive AML patients. When possible, results on the leukemic fraction were normalized to the cognate lymphocyte population (relative priming and BCL-2 dependence). Samples from 97 (89.8%) of the 108 referred patients were successfully processed. Relative priming and BCL-2 dependence could be determined in 62 (67.4%) and 67 (62.0%) samples, respectively. Absolute mitochondrial priming was lower in patients having previously failed intensive chemotherapy compared to chemotherapy-naïve patients (p = 0.01), but its prognostic impact was limited. Conversely, relative BCL-2 independence tended to predict worse EFS (HR = 2.51, p = 0.07) and OS (HR = 2.79, p = 0.10) independently of adverse genetic risk. Our results show that simplified BH3 profiling can be prospectively assessed in AML patients but that its prognostic use may require internal normalization. Future studies should compare its relevance with other functional assays such as ex vivo drug testing or BH3 protein expression.
Insights
Simplified BH3 profiling shows promise for assessing acute myeloid leukemia (AML) patient mitochondrial priming and BCL-2 dependence. Relative BCL-2 independence may indicate poorer outcomes, suggesting normalization is key for prognostic use.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- BH3 profiling assesses mitochondrial priming and dependence on anti-apoptotic proteins like BCL-2 in cancer.
- Previous prognostic studies in acute myeloid leukemia (AML) used archived samples with variable molecular genetics.
- A simplified, flow-based assay offers a feasible approach for prospective BH3 profiling in AML.
Purpose of the Study:
- To evaluate the feasibility of a simplified flow-based BH3 profiling assay in consecutive AML patients.
- To determine absolute and relative mitochondrial priming and BCL-2 dependence.
- To assess the prognostic impact of these parameters in AML.
Main Methods:
- A single-center feasibility study involving 108 consecutive AML patients.
- Simplified flow-based BH3 profiling assay to measure mitochondrial priming and BCL-2 dependence.
- Normalization of leukemic cell results to lymphocyte populations where possible.
Main Results:
- BH3 profiling was successfully performed on 97 (89.8%) patients.
- Relative priming and BCL-2 dependence were determined in 67.4% and 62.0% of samples, respectively.
- Absolute mitochondrial priming was lower in pre-treated patients (p=0.01), but prognostic impact was limited.
- Relative BCL-2 independence trended towards worse event-free survival (EFS) and overall survival (OS) (HR=2.51 and 2.79, respectively).
Conclusions:
- Simplified BH3 profiling is prospectively feasible in AML.
- Internal normalization (relative measurements) may be crucial for prognostic utility.
- Further studies are needed to compare BH3 profiling with other functional assays like ex vivo drug testing or protein expression analysis.
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