Next-Generation Sequencing in Diffuse Large B-Cell Lymphoma Highlights Molecular Divergence and Therapeutic Opportunities: a LYSA Study

Clin Cancer Res. 2016 Jun 15;22(12):2919-28. doi: 10.1158/1078-0432.CCR-15-2305. Epub 2016 Jan 27.


Purpose: Next-generation sequencing (NGS) has detailed the genomic characterization of diffuse large B-cell lymphoma (DLBCL) by identifying recurrent somatic mutations. We set out to design a clinically feasible NGS panel focusing on genes whose mutations hold potential therapeutic impact. Furthermore, for the first time, we evaluated the prognostic value of these mutations in prospective clinical trials.

Experimental design: A Lymphopanel was designed to identify mutations in 34 genes, selected according to literature and a whole exome sequencing study of relapsed/refractory DLBCL patients. The tumor DNA of 215 patients with CD20(+)de novo DLBCL in the prospective, multicenter, and randomized LNH-03B LYSA clinical trials was sequenced to deep, uniform coverage with the Lymphopanel. Cell-of-origin molecular classification was obtained through gene expression profiling with HGU133+2.0 Affymetrix GeneChip arrays.

Results: The Lymphopanel was informative for 96% of patients. A clear depiction of DLBCL subtype molecular heterogeneity was uncovered with the Lymphopanel, confirming that activated B-cell-like (ABC), germinal center B-cell like (GCB), and primary mediastinal B-cell lymphoma (PMBL) are frequently affected by mutations in NF-κB, epigenetic, and JAK-STAT pathways, respectively. Novel truncating immunity pathway, ITPKB, MFHAS1, and XPO1 mutations were identified as highly enriched in PMBL. Notably, TNFAIP3 and GNA13 mutations in ABC patients treated with R-CHOP were associated with significantly less favorable prognoses.

Conclusions: This study demonstrates the contribution of NGS with a consensus gene panel to personalized therapy in DLBCL, highlighting the molecular heterogeneity of subtypes and identifying somatic mutations with therapeutic and prognostic impact. Clin Cancer Res; 22(12); 2919-28. ©2016 AACRSee related commentary by Lim and Elenitoba-Johnson, p. 2829.

Publication types

  • Multicenter Study
  • Randomized Controlled Trial

MeSH terms

  • Antibodies, Monoclonal, Murine-Derived / therapeutic use
  • Antineoplastic Agents / therapeutic use*
  • Antineoplastic Combined Chemotherapy Protocols / therapeutic use*
  • B-Lymphocytes / immunology
  • B-Lymphocytes / pathology
  • Cell Cycle Proteins / genetics
  • Cyclophosphamide / therapeutic use
  • DNA-Binding Proteins / genetics
  • Doxorubicin / therapeutic use
  • Exome Sequencing
  • Exportin 1 Protein
  • GTP-Binding Protein alpha Subunits, Gq-G11 / genetics
  • Gene Expression Profiling
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Karyopherins / genetics
  • Lymphoma, Large B-Cell, Diffuse / drug therapy*
  • Lymphoma, Large B-Cell, Diffuse / genetics*
  • Molecular Targeted Therapy / methods*
  • Oncogene Proteins / genetics
  • Phosphotransferases (Alcohol Group Acceptor) / genetics
  • Precision Medicine / methods*
  • Prednisone / therapeutic use
  • Prospective Studies
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Rituximab
  • Tumor Necrosis Factor alpha-Induced Protein 3 / genetics
  • Vincristine / therapeutic use


  • Antibodies, Monoclonal, Murine-Derived
  • Antineoplastic Agents
  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Karyopherins
  • MFHAS1 protein, human
  • Oncogene Proteins
  • R-CHOP protocol
  • Receptors, Cytoplasmic and Nuclear
  • Rituximab
  • Vincristine
  • Doxorubicin
  • Cyclophosphamide
  • Phosphotransferases (Alcohol Group Acceptor)
  • Inositol 1,4,5-trisphosphate 3-kinase
  • TNFAIP3 protein, human
  • Tumor Necrosis Factor alpha-Induced Protein 3
  • GNA14 protein, human
  • GTP-Binding Protein alpha Subunits, Gq-G11
  • Prednisone