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Published on: December 28, 2015
Accurate Classification of Germinal Center B-Cell-Like/Activated B-Cell-Like Diffuse Large B-Cell Lymphoma Using a
Sylvain Mareschal1, Philippe Ruminy1, Cristina Bagacean2
1INSERM U918, Centre Henri Becquerel, Institute for Research and Innovation in Biomedicine, University of Rouen, Rouen, France.
Insights
A new classifier accurately distinguishes between germinal center B-cell-like and activated B-cell-like diffuse large B-cell lymphomas. This method aids in treatment decisions for non-Hodgkin lymphoma patients.
Area of Science:
- Hematology
- Molecular Biology
- Oncology
Background:
- Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma.
- DLBCL is subtyped into germinal center B-cell-like (GCB) and activated B-cell-like (ABC) subtypes.
- Accurate differentiation of DLBCL subtypes is challenging in routine diagnostics, hindering targeted therapy development.
Purpose of the Study:
- To develop a simple, rapid, and accurate classifier for DLBCL subtypes.
- To validate the classifier's performance using independent cohorts and formalin-fixed, paraffin-embedded samples.
- To assess the clinical utility of the classifier in patient stratification and treatment decisions.
Main Methods:
- Development of a classifier using reverse transcriptase multiplex ligation-dependent probe amplification (RT-MLPA) assay and 14 gene signatures.
- Validation of the classifier against a gold standard (Affymetrix U133+2) and a mid-throughput gene expression profiling method.
- Inclusion of prognostic factors like MYC/BCL2 co-expression in the assay.
Main Results:
- High concordance (95% CI, 92%-100%) with the gold standard in a validation cohort.
- Accurate classification of 93% (95% CI, 82%-98%) of samples in an independent series.
- Demonstrated significant differences in overall and progression-free survival between subtypes in a 135-patient cohort.
Conclusions:
- The developed RT-MLPA-based classifier provides a sensitive, rapid, and reliable method for DLBCL subtype differentiation.
- The assay's flexibility and ease of implementation in clinical settings facilitate patient stratification for targeted therapies.
- This approach can significantly aid in optimizing treatment strategies for DLBCL patients.
Abstract:
Diffuse large B-cell lymphoma, the most common non-Hodgkin lymphoma, is subdivided into germinal center B-cell-like and activated B-cell-like subtypes. Unfortunately, these lymphomas are difficult to differentiate in routine diagnosis, impeding the development of treatments. Patients with these lymphomas can benefit from specific therapies. We therefore developed a simple and rapid classifier based on a reverse transcriptase multiplex ligation-dependent probe amplification assay and 14 gene signatures. Compared with the Affymetrix U133+2 gold standard, all 46 samples (95% CI, 92%-100%) of a validation cohort classified by both techniques were attributed to the expected subtype. Similarly, 93% of the 55 samples (95% CI, 82%-98%) of a second independent series characterized with a mid-throughput gene expression profiling method were classified correctly. Unclassifiable sample proportions reached 13.2% and 13.8% in these cohorts, comparable with the frequency originally reported. The developed assay was also sensitive enough to obtain reliable results from formalin-fixed, paraffin-embedded samples and flexible enough to include prognostic factors such as MYC/BCL2 co-expression. Finally, in a series of 135 patients, both overall (P = 0.01) and progression-free (P = 0.004) survival differences between the two subtypes were confirmed. Because the multiplex ligation-dependent probe amplification method is already in use and requires only common instruments and reagents, it could easily be applied to clinical trial patient stratification to help in treatment decisions.
