Determination of Molecular Subtypes of Diffuse Large B-Cell Lymphoma Using a Reverse Transcriptase Multiplex Ligation-Dependent Probe Amplification Classifier

Abstract

Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma. It includes three major subtypes termed germinal center B-cell-like, activated B-cell-like, and primary mediastinal B-cell lymphoma. With the emergence of novel targeted therapies, accurate methods capable of interrogating this cell-of-origin classification should soon become essential in the clinics. To address this issue, we developed a novel gene expression profiling DLBCL classifier based on reverse transcriptase multiplex ligation-dependent probe amplification. This assay simultaneously evaluates the expression of 21 markers, to differentiate primary mediastinal B-cell lymphoma, activated B-cell-like, germinal center B-cell-like, and also Epstein-Barr virus-positive DLBCLs. It was trained using 70 paraffin-embedded biopsies and validated using >160 independent samples. Compared with a reference classification established from Affymetrix U133 + 2 data, reverse transcriptase multiplex ligation-dependent probe amplification classified 85.0% samples into the expected subtype, comparing favorably with current diagnostic methods. This assay also proved to be highly efficient in detecting the MYD88 L265P mutation, even in archival paraffin-embedded tissues. This reliable, rapid, and cost-effective method uses common instruments and reagents and could thus easily be implemented into routine diagnosis workflows, to improve the management of these aggressive tumors.

Incorporation of digital gene expression profiling for cell-of-origin determination (Lymph2Cx testing) into the routine work-up of diffuse large B cell lymphoma

“…Indeed, a number of other groups have also developed molecular methods to determine COO in DLBCL. For example, Mareschal et al developed a gene-expression assay for COO determination in DLBCL based on a reverse transcriptase-multiplex ligation-dependent probe amplification (RT-MLPA) and also recently expanded their assay to include the classification of primary mediastinal large B cell lymphoma [22,23]. In addition, Collie et al developed a novel multiplex single-tube gene expression assay on the ICEPlex® system which could differentiate between GCB and ABC DLBCL subtypes in FFPE tissue specimens [24].…”

STAT3-coordinated migration facilitates the dissemination of diffuse large B-cell lymphomas

“…Risk stratification was conducted using the International Prognostic Index (IPI) score. From the 73 DLBCL patients, 40 available patient samples were subjected to an 8-gene expression panel analysis to determine the GCB and ABC subtypes as reported previously 42 – 44 . RNA was extracted from FFPE tissue (ReliaPrep™ FFPE Total RNA Miniprep System, Promega) and used for cDNA synthesis.…”

Cytological diagnosis of vitreoretinal lymphomas: A case series

“…Treatment depends on the age and general condition of the patient, and whether they are associated with cerebral lymphoma. Treatments consist of systemic chemotherapy or local treatment (intra‐vitreous chemotherapy or ocular radiotherapy) . Vitrectomy is also described as an adjunct treatment to systemic therapy for VRL …”