Reliable subtype classification of diffuse large B-Cell lymphoma samples from GELA LNH2003 trials using the Lymph2Cx gene expression assay (original) (raw)
Blood, 2004
Diffuse large B-cell lymphoma (DLBCL) can be divided into prognostically important subgroups with germinal center Bcell-like (GCB), activated B-cell-like (ABC), and type 3 gene expression profiles using a cDNA microarray. Tissue microarray (TMA) blocks were created from 152 cases of DLBCL, 142 of which had been successfully evaluated by cDNA microarray (75 GCB, 41 ABC, and 26 type 3). Sections were stained with antibodies to CD10, bcl-6, MUM1, FOXP1, cyclin D2, and bcl-2. Expression of bcl-6 (P < .001) or CD10 (P ؍ .019) was associated with better overall survival (OS), whereas expression of MUM1 (P ؍ .009) or cyclin D2 (P < .001) was associated with worse OS. Cases were subclassified using CD10, bcl-6, and MUM1 expression, and 64 cases (42%) were considered GCB and 88 cases (58%) non-GCB. The 5-year OS for the GCB group was 76% compared with only 34% for the non-GCB group (P < .001), which is similar to that reported using the cDNA microarray. Bcl-2 and cyclin D2 were adverse predictors in the non-GCB group. In multivariate analysis, a high International Prognostic Index score (3-5) and the non-GCB phenotype were independent adverse predictors (P < .0001). In summary, immunostains can be used to determine the GCB and non-GCB subtypes of DLBCL and predict survival similar to the cDNA microarray. (Blood. 2004;103:275-282)
Reproducibility of Gene Expression Signatures in Diffuse Large B-Cell Lymphoma
Cancers, 2022
Multiple gene expression profiles have been identified in diffuse large B-cell lymphoma (DLBCL). Besides the cell of origin (COO) classifier, no signatures have been reproduced in independent studies or evaluated for capturing distinct aspects of DLBCL biology. We reproduced 4 signatures in 175 samples of the HOVON-84 trial on a panel of 117 genes using the NanoString platform. The four gene signatures capture the COO, MYC activity, B-cell receptor signaling, oxidative phosphorylation, and immune response. Performance of our classification algorithms were confirmed in the original datasets. We were able to validate three of the four GEP signatures. The COO algorithm resulted in 94 (54%) germinal center B-cell (GCB) type, 58 (33%) activated B-cell (ABC) type, and 23 (13%) unclassified cases. The MYC-classifier revealed 77 cases with a high MYC-activity score (44%) and this MYC-high signature was observed more frequently in ABC as compared to GCB DLBCL (68% vs. 32%, p < 0.00001). T...
British Journal of Haematology
We assessed the concordance between immunohistochemistry (IHC) and gene expression profiling (GEP) for determining diffuse large B-cell lymphoma (DLBCL) cell of origin (COO) in the phase III PHOENIX trial of rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) with or without ibrutinib. Among 910/1114 screened patients with nongerminal centre B-cell-like (non-GCB) DLBCL by IHC, the concordance with GEP for non-GCB calls was 82•7%, with 691 (75•9%) identified as activated B-cell-like (ABC), and 62 (6•8%) as unclassified. Among 746/837 enrolled patients with verified non-GCB DLBCL by This trial was funded by Janssen Research and Development. The authors would like to thank Dr Anas Younes' contributions towards the PHOENIX study and all patients included in this analysis. Writing assistance was provided by
Clinical Lymphoma Myeloma and Leukemia, 2011
Diffuse large B-cell lymphoma (DLBCL) as described by the World Health Organization (WHO) 2008 classification is the most frequent lymphoma subtype of the aggressive Non-Hodgkin's lymphomas (NHL). 1 The addition of rituximab to CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone)-containing regimens (R-CHOP) highly improves outcome. However, survival of both elderly and young patients with NHL is still unsatisfactory, making improvement of current therapeutic strategies necessary. 2-12 One of the most promising strategies may be tailoring the therapy to the clinical and biologic factors of patients and disease, which influence outcome. To this end, the International Prognostic Index (IPI) up to now has been the most powerful tool for predicting clinical outcome of patients with DLBCL. However, the IPI reflects a mixture of underlying biologic differences and thus has its limits with respect to the aim of personalized therapy. Therefore, novel biologic prognostic markers reflecting the heterogeneous biology of DLBCL have been investigated with the ultimate goal of finding DLBCL subtypes with different prognosis. 13 Gene expression profiling (GEP) using oligonucleotide arrays indeed confirmed the existence of subgroups of patients, treated either with CHOP or CHOP-like
International journal of molecular epidemiology and genetics, 2011
Gene expression profiling studies have distinguished diffuse large B-cell lymphomas (DLBCLs) by cell of origin, with distinct pathogenetic mechanisms and prognosis. We attempted to identify DLBCL molecular subtypes in an epidemiologic study of 214 DLBCL patients diagnosed during 1998-2000 with archival tissues to investigate etiology. Immunohistochemical staining for CD10, BCL6, LMO2, MUM1/IRF4, and BCL2 and fluorescence in situ hybridization for t(14;18) were conducted, with ≥93% blinded duplicate agreement. CD10, LMO2, and BCL2 expression was similar to previous reports (32%, 44%, and 44% of DLBCLs, respectively), but BCL6 and MUM1/IRF4 expression was lower than expected (29% and 5%, respectively). We classified 112/214 (52%) cases as germinal center B-cell-like DLBCL (GCB-DLBCL; Hans et al., Blood 2004; CD10+ or CD10-/BCL6+/MUM1-), with no difference in prognosis compared with non-GCB-DLBCL (Cox regression, P=0.48). Comparing other GCB correlates, LMO2 expression and t(14;18) wer...
Hematological Oncology, 2019
Introduction: Sequencing studies identified mutational drivers in diffuse large B cell lymphoma (DLBCL), capturing outcome difference in previously unrecognized patient subsets. However, the lack of routine-applicable genomic approaches limits translation of such information to the clinic. Currently, molecular prognostication consists in cell of origin (COO) determination by the Lymph2Cx NanoString assay. We recently developed two independent prognostic signatures incorporating genes reflecting the COO, the activation of pivotal oncogenic pathways, and the composition of tumor microenvironment (TME). We aimed this study at examining the prognostic strength of a model combining the performance of each signature and developing a comprehensive NanoString assay rapidly transferable to the clinic for prognostic purposes. Methods: The expression of the genes was measured by the NanoString nCounter Analysis System using customized probes for 73 genes, including 15 COO genes, 6 additional oncogenic genes (MYC, BCL-2, NFKBIA, PIK3CA, PTEN, STAT3), 47 TME genes, and 5 housekeeping genes. The analysis was performed on 175 newly
Clinical Cancer Research, 2013
Purpose: The opportunity to improve therapeutic choices on the basis of molecular features of the tumor cells is on the horizon in diffuse large B-cell lymphoma (DLBCL). Agents such as bortezomib exhibit selective activity against the poor outcome activated B-cell type (ABC) DLBCL. In order for targeted therapies to succeed in this disease, robust strategies that segregate patients into molecular groups with high reliability are needed. Although molecular studies are considered gold standard, several immunohistochemistry (IHC) algorithms have been published that claim to be able to stratify patients according to their cell-of-origin and to be relevant for patient outcome. However, results are poorly reproducible by independent groups. Experimental Design: We investigated nine IHC algorithms for molecular classification in a dataset of DLBCL diagnostic biopsies, incorporating immunostaining for CD10, BCL6, BCL2, MUM1, FOXP1, GCET1, and LMO2. IHC profiles were assessed and agreed among three expert observers. A consensus matrix based on all scoring combinations and the number of subjects for each combination allowed us to assess reliability. The survival impact of individual markers and classifiers was evaluated using Kaplan-Meier curves and the log-rank test. Results: The concordance in patient's classification across the different algorithms was low. Only 4% of the tumors have been classified as germinal center B-cell type (GCB) and 21% as ABC/non-GCB by all methods. None of the algorithms provided prognostic information in the R-CHOP (rituximab plus cyclophosphamide-adriamycin-vincristine-prednisone)-treated cohort. Conclusion: Further work is required to standardize IHC algorithms for DLBCL cell-of-origin classification for these to be considered reliable alternatives to molecular-based methods to be used for clinical decisions. Clin Cancer Res; 1-10. Ó2013 AACR.
British journal of haematology, 2017
The study included 1848 diffuse large B-cell lymphoma (DLBCL)patients treated with chemotherapy/rituximab. The aims were to validate the National Comprehensive Cancer Network International Prognostic Index (NCCN-IPI) and explore the effect of adding high Beta-2 microglobulin (β2M), primary extranodal presentation and intense treatment to the NCCN-IPI variables in order to develop an improved index. Comparing survival curves, NCCN-IPI discriminated better than IPI, separating four risk groups with 5-year overall survival rates of 93%, 83%, 67% and 49%, but failing to identify a true high-risk population. For the second aim the series was split into training and validation cohorts: in the former the multivariate model identified age, lactate dehydrogenase, Eastern Cooperative Oncology Group performance status, Stage III-IV, and β2M as independently significant, whereas the NCCN-IPI-selected extranodal sites, primary extranodal presentation and intense treatments were not. These result...