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Determination of Molecular Subtypes of Diffuse Large B-Cell Lymphoma Using a Reverse Transcriptase Multiplex
Victor Bobée1, Philippe Ruminy2, Vinciane Marchand2
1INSERM U1245, UNIROUEN, University of Normandie, Rouen, France; Department of Biological Hematology, Rouen University Hospital, Rouen, France.
The Journal of Molecular Diagnostics : JMD
|October 22, 2017
Summary
A new gene expression profiling assay accurately classifies Diffuse Large B-cell Lymphoma (DLBCL) subtypes using multiplex ligation-dependent probe amplification. This cost-effective method aids in managing aggressive tumors and detecting mutations in archival tissues.
Area of Science:
- Hematology
- Oncology
- Molecular Diagnostics
Background:
- Diffuse Large B-cell Lymphoma (DLBCL) is the most prevalent non-Hodgkin lymphoma, with distinct subtypes requiring precise classification for effective treatment.
- Emerging targeted therapies necessitate accurate methods for determining DLBCL cell-of-origin classification in clinical settings.
Purpose of the Study:
- To develop and validate a novel gene expression profiling classifier for DLBCL subtypes using reverse transcriptase multiplex ligation-dependent probe amplification (RT-MLPA).
- To assess the assay's efficiency in differentiating between germinal center B-cell-like, activated B-cell-like, primary mediastinal B-cell lymphoma, and Epstein-Barr virus-positive DLBCL.
Main Methods:
- Development of an RT-MLPA assay evaluating 21 gene expression markers simultaneously.
- Training the classifier on 70 paraffin-embedded DLBCL biopsies and validation on over 160 independent samples.
- Comparison with a reference classification based on Affymetrix U133+2 data.
Main Results:
- The RT-MLPA classifier achieved 85.0% accuracy in subtype classification compared to the reference standard.
- The assay demonstrated high efficiency in detecting the MYD88 L265P mutation, including in archival paraffin-embedded tissues.
- Performance favorably compared with current diagnostic methods.
Conclusions:
- The developed RT-MLPA assay provides a reliable, rapid, and cost-effective method for DLBCL subtyping.
- The assay's ease of implementation using common laboratory equipment facilitates integration into routine diagnostic workflows.
- Improved DLBCL classification has the potential to enhance the management of these aggressive hematologic malignancies.
