Neither Germinal Center (GC) vs Non-Germinal Center (Non-GC) Phenotype nor FOXP1 Expression Correlate with Outcome in AIDS-Associated Diffuse Large B-Cell Lymphoma (DLBCL): Study of Patients from AIDS Malignancies Consortium Trials 010 and 034.

Abstract Background: Germinal center (GC) and non-germinal center (non-GC) gene expression profiles correlate with survival in immunocompetent DLBCL patients. Phenotypic expression patterns of CD10, BCL6, MUM1 and CD138 are surrogates for genetic studies with comparable survival data; CD10+, BCL6+/−, MUM1- defines GC while CD10-, BCL6-, MUM1+/− identifies the non-GC phenotype with poorer prognosis. Expression of FOXP1, a transcription factor differentially expressed in resting and activated B cells, and PRDM1/BLIMP1, a regulator of terminal B cell differentiation, are also adverse prognostic markers for DLBCL in immunocompetent patients. AIDS-associated DLBCLs from uniformly treated HIV+ patients in AMC010 (CHOP vs. CHOP-rituxan) and AMC034 (EPOCH vs. EPOCH-rituxan) were examined to determine if the GC vs. non-GC phenotype, FOXP1 expression and/or BLIMP1 expression are prognostic in this patient population. Design: Slides of 32 and 30 AIDS-associated DLBCLs from AMC010 (closed) and AMC034 (in analysis) patients, respectively, were available for FOXP1, BLIMP1, CD10, BCL6, MUM1, BCL2, Ki-67 immunohistochemistry and in situ hybridization for EBV (EBER). Antigen expression by >20% tumor cells (>10% for BLIMP1) was considered positive. GC phenotype was defined as CD10+, BCL6+/−, MUM1- while the non-GC cases were CD10-, BCL6-, MUM1+/−. Overall survival (OS) based on GC vs. non-GC phenotype was examined. FOXP1 expression was correlated with survival; BCL2, Ki-67 expression; and EBV status; BLIMP1 expression was correlated with survival in a subgroup of patients from AMC 034. Results: Of the 62 cases, 59% were MUM1, 60% BCL6, 53% CD10, 59% BCL2, 49% FOXP1, 67% BLIMP1 and 30% EBER positive. 19 (58%) cases were classified as GC and 14 as non-GC. No mean OS difference between GC and non-GC groups (p=0.74) or FOXP1+ and FOXP1- cases (p=0.8; t-test) in the AMC 010 patients; BLIMP1 expression did not correlate with survival in the subgroup of AMC 034 patients (p=0.4). GC vs. non-GC phenotype did not correlate with FOXP1 expression (p=0.1) or BLIMP1 expression (p=0.4). In addition, FOXP1 expression did not correlate with EBV positivity, BCL2, MUM1, BCL6 or CD10 expression or proliferation rate based on Ki67 (chi-square). Conclusions: AIDS-associated DLBCLs can be classified as GC and non-GC cases, but this classification does not appear to correlate with prognosis/OS in uniformly treated HIV-positive patients. Furthermore, in contrast with immunocompetent DLBCLs, FOXP1 expression also does not correlate with OS in this patient population; similarly BLIMP1 expression also does not correlate with survival in the HIV+ patient population. In addition, GC vs non-GC phenotype did not correlate with FOXP1 or BLIMP1 expression, while FOXP1 expression did not correlate with prognostic markers BCL2/Ki67 or EBV status. Thus, prognostic markers useful in immunocompetent patients with DLBCL may not be relevant for HIV positive patients, suggesting that patient immune status rather than tumor biology may be more important in predicting patient outcome.

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Addition of Rituximab to CHOP Regimen Improves Clinical Outcome in Non-Germinal Center Type Diffuse Large B-Cell Lymphoma.

Blood ◽

10.1182/blood.v108.11.4725.4725 ◽

2006 ◽

Vol 108 (11) ◽

pp. 4725-4725

Author(s):

Masahiro Yokoyama ◽

Daisuke Ennishi ◽

Kyoko Ueda ◽

Makoto Kodaira ◽

Shuhei Yamada ◽

...

Keyword(s):

Clinical Outcome ◽

B Cell ◽

Germinal Center ◽

Cell Lymphoma ◽

Expression Patterns ◽

B Cell Lymphoma ◽

Chop Regimen ◽

Large B Cell Lymphoma ◽

Type 3 ◽

Large B Cell

Abstract Background: In recent years, diffuse large B-cell lymphoma (DLBCL) has been classified into the germinal center B-cell (GC) type, the activated B-cell (ABC) type, and the type 3 using global gene expression profiling or immunohistochemical staining. It has been reported that the GC type DLBCL showed significantly longer survival than the non-GC (ABC and the type 3) type DLBCL treated with CHOP or CHOP like regimen not using rituximab. Methods: We analyzed retrospectively the prognosis between the GC and non-GC types of DLBCL treated with R-CHOP regimen. All 50 patients with DLBCL, diagnosed between July 2003 and July 2005 were included in this study. The pathology was reviewed by hematopathologist and confirmed to be de novo DLBCL according to the WHO classification. Patients with primary CNS- and post-transplant lymphomas were excluded. GC type or non-GC type DLBCL was determined by immunohistochemistry such as the expression patterns of CD10, BCL-6, and IRF-4 (MUM1). All patients were initially treated with six cycles of R-CHOP regimen consisting of rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone. If we evaluated partial response after six cycles of R-CHOP, the patients have added radiation therapy. Results: The patients consisted of 30 GC type and 20 non-GC type DLBCL with a median age of 61.0 yr (range 31–83 yr). The median follow up of surviving patients was 24 months. CR rate between GC and non-GC types were 57.0% vs. 75.0%, p=0.186, and overall response rate were 87.0% vs. 90.0%, p=0.929, respectively. The median of progression free survival was 17.3 months vs. 19.6 months, p=0.80. There is no statistical significance difference between two groups. Conclusion: These results suggest that addition of rituximab to CHOP regimen improves clinical outcome of non-GC type DLBCL as well as GC type DLBCL.

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Distinct Expression Patterns of microRNAs in Activated B Cell (ABC) and Germinal Center B (GC) Subtypes of Diffuse Large B Cell Lymphoma (DLBLC).

Blood ◽

10.1182/blood.v110.11.562.562 ◽

2007 ◽

Vol 110 (11) ◽

pp. 562-562

Author(s):

Jean-Noel Bastie ◽

Jean-Philippe Jais ◽

Thierry Molina ◽

Emmanuelle Come ◽

Bertrand Coiffier ◽

...

Keyword(s):

B Cell ◽

Mirna Expression ◽

Germinal Center ◽

Cell Lymphoma ◽

Expression Patterns ◽

B Cell Lymphoma ◽

Rt Pcr ◽

Large B Cell Lymphoma ◽

Micro Rnas ◽

Large B Cell

Abstract The Micro RNAs (miRNAs) are small non-coding, single-stranded RNAs that regulate the expression of genes by hybridizing the mRNA target with complementary sequences that are followed by translational repression, mRNa cleavage or destabilization. There is evidence that miRNA play an important role in carcinogenesis. Aberrant miRNA expression has been found in a variety of human malignancies including B-cell leukemias and lymphomas. The purpose of this study is to determine the miRNA expression patterns in DLBCL and their relationship with clinical characteristics and outcome. We used high throughput quantitative RT-PCR technology (Taqman Low density Arrays) to analyze the expression profile of 365 different mature miRNA in 12 Diffuse Large B-Cell Lymphoma (DLBCL) samples which had been previously characterized at the transcriptional level with Affymetrix HU133A micro-arrays. MiR-155 expression was significantly lower in the 6 samples with a Germinal Center (GC) mRNA profile than in the 6 samples with an Activated B-Cell (ABC) profile. Expression of miR-155 and of different miRNA involved in lymphoid differentiation (miR-181a, miR127) or tumour pathogenesis (cluster miR 17–92) were further evaluated in a series of 64 patients with DLBCL treated with Rituximab associated with chemotherapy (R-CHOP). 28 patients had been enrolled in the LNH98.5 GELA trial between February 1998 and February 2000 and 36 were treated with R-CHOP in GELA centers after the closure of the LNH98-5 trial, between November 2000 and June 2004. The median follow up of these patients is 69 months. Patients median age at diagnosis was 69 years (range 59–82) and 28 patients presented with an International Prognostic Index (IPI) score of more than 3. Mature miRNA expression was determined by quantitative RT-PCR with Applied Biosystems specific miRNA primer and probe sets and normalized to U6 small nuclear RNA expression. MiR-155 expression was significantly higher in patients with a lymphoma of the ABC subtype, defined on the basis of the transcriptional profile (mean delta Ct = − 3.9 in the 41 ABC samples, mean delta Ct = − 1.67 in the 23 GC samples, p<0.00001) and significantly higher in patients with a high (4 or 5) IPI score (p=0.02). A high miR-155 expression was associated with a trend towards a poorer overall survival. The expression of miR-127, miR-181a and the cluster 17–92 were not correlated with clinical outcome. These analyses are currently being extended to a larger series of patients in order to determine whether other miRNA can be used to classify DLBCL samples into ABC and GC subtypes and whether some miRNA have prognostic significance in the era of treatments combining Rituximab and chemotherapy.

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Tumor-specific and Proliferation-specific Gene Expression Typifies Murine Transgenic B Cell Lymphomagenesis

Journal of Biological Chemistry ◽

10.1074/jbc.m605870200 ◽

2006 ◽

Vol 282 (7) ◽

pp. 4803-4811 ◽

Cited By ~ 27

Author(s):

Marc E. Lenburg ◽

Anupama Sinha ◽

Douglas V. Faller ◽

Gerald V. Denis

Keyword(s):

Gene Expression ◽

Cell Cycle ◽

B Cell ◽

Cell Cycle Progression ◽

Cellular Proliferation ◽

Expression Profiles ◽

Expression Patterns ◽

B Cell Lymphoma ◽

Specific Pattern ◽

Specific Gene

The dual bromodomain protein Brd2 is closely related to the basal transcription factor TAFII250, which is essential for cyclin A transactivation and mammalian cell cycle progression. In transgenic mice, constitutive lymphoid expression of Brd2 causes a malignancy most similar to human diffuse large B cell lymphoma. We compare the genome-wide transcriptional expression profiles of these lymphomas with those of proliferating and resting normal B cells. Transgenic tumors reproducibly show differential expression of a large number of genes important for cell cycle control and lymphocyte biology; expression patterns are either tumor-specific or proliferation-specific. Several of their human orthologs have been implicated in human lymphomagenesis. Others correlate with human disease survival time. BRD2 is underexpressed in some subtypes of human lymphoma and these subtypes display a number of similarities to the BRD2-mediated murine tumors. We illustrate with a high degree of detail that cancer is more than rampant cellular proliferation, but involves the additional transcriptional mobilization of many genes, some of them poorly characterized, which show a tumor-specific pattern of gene expression.

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BCL2/Ki-67 index predict survival in germinal center B-cell-like diffuse large B-cell lymphoma

Oncology Letters ◽

10.3892/ol.2017.6577 ◽

2017 ◽

Vol 14 (3) ◽

pp. 3767-3773 ◽

Cited By ~ 9

Author(s):

Yun-Long Tang ◽

Yan Zhou ◽

Ling-Ling Cheng ◽

Yong-Zhong Su ◽

Chun-Bin Wang

Keyword(s):

B Cell ◽

Germinal Center ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Ki 67 ◽

Large B Cell Lymphoma ◽

Germinal Center B Cell ◽

Large B Cell

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MicroRNA Expression Profiling Characterize Diffuse Large B-Cell Lymphomas and Follicular Lymphomas.

Blood ◽

10.1182/blood.v110.11.3186.3186 ◽

2007 ◽

Vol 110 (11) ◽

pp. 3186-3186

Author(s):

Anja Roehle ◽

Kai P. Hoefig ◽

Dirk Repsilber ◽

Christoph Thorns ◽

Kai O. Wesche ◽

...

Keyword(s):

Lymph Nodes ◽

B Cell ◽

Mirna Expression ◽

Germinal Center ◽

Expression Profiles ◽

Expression Patterns ◽

Mirna Expression Profiles ◽

Proliferation And Differentiation ◽

Follicular Lymphomas ◽

Large B Cell

Abstract A new and powerful tool for classification and understanding the biology of human lymphomas is microRNA (miRNA) expression profiling. MiRNAs comprise a class of approximately 1000 RNA species, which are thought to directly regulate the expression of ∼30% of the transcripts of the human genome on a posttranscriptional level. An important regulatory role of miRNAs has been implicated in biological processes such as cell proliferation and differentiation, fat metabolism, insulin secretion and hematopoiesis. Recent studies indicate that miRNAs are mechanistically involved in the development of various human malignancies and therefore represent a promising new class of biomarkers. We evaluated the miRNA expression profiles of 58 Diffuse Large B-Cell Lymphomas (DLBCL), 46 Follicular Lymphomas (FL) and 7 lymph nodes with chronic lymphadenitis. A quantitative PCR-based method was used to determine the expression levels of 157 miRNA species. The analysis was performed exclusively on FFPE tissues, which are traditionally inapt to molecular analysis. It was possible to clearly distinguish malignant and non-malignant tissues by miRNA expression patterns. We found 34 differentially expressed miRNAs by comparing DLBCL and lymph nodes, 25 miRNAs by comparing FL and lymph nodes and 55 by comparing DLBCL and FL. It was noticeable that the miRNA expression profiles of FL and lymph nodes are more similar compared to miRNA expression profiles of DLBCL. We were also interested to find out whether it is possible to classify the DLBCL cases in germinal center-like DLBCL (GCB-DLBCL) and Non-germinal center-like DLBCL (Non-GCB-DLBCL) based on miRNA expression patterns. We used the immunohistochemical classification, published by Hans et al., to identify both subgroups (Hans CP, Weisenburger DD et al. Confirmation of the molecular classification of diffuse large B-cell lymphoma by immunohistochemistry using a tissue microarray. Blood. 2004; 103:275–282). Comparing miRNA expression profiles of GCB and Non-GCB cases of DLBCL, 8 miRNAs were found to be differentially expressed using the partial least squares discriminant analysis (PLS-DA). However, using a cross-validation approach it was not possible to predict the GCB status on the basis of miRNA expression patterns. Similar to Landgraf et al. we found a reduced miR-150 expression level in DLBCL in comparison to other closely related germinal center malignancies (Landgraf P, Ruscu M et al. A mammalian microRNA expression atlas based on small RNA library sequencing. Cell. 2007; 129:1401–1414). To determine the function of miR-150, Ramos cells were transfected with anti-miR-150 and an analysis of putative target proteins was carried out. It was possible to show a regulation of transcription factor v-myb/c-myc, which plays an important role in the control of proliferation and differentiation of hematopoietic progenitor cells, by miR-150. Moreover, miRNA profiles of DLBCL from a randomized trail will be compared to survival data in the future.

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Phylogenetic Evolution in Refractory/Relapsed Diffuse Large B-Cell Lymphoma

Blood ◽

10.1182/blood.v120.21.418.418 ◽

2012 ◽

Vol 120 (21) ◽

pp. 418-418

Author(s):

Philippe Ruminy ◽

Vinciane Rainville ◽

Sylvain Mareschal ◽

Pascaline Etancelin ◽

Berangere Joly ◽

...

Keyword(s):

B Cell ◽

Germinal Center ◽

Phylogenetic Trees ◽

Somatic Mutations ◽

Cell Lymphoma ◽

Expression Profiles ◽

Gene Expression Profiles ◽

B Cell Lymphoma ◽

Large B Cell Lymphoma ◽

Selection Of

Abstract Abstract 418 Diffuse large B-cell lymphoma (DLBCL) is the most common non-hodgkin lymphoma. It can be subdivided into two major subtypes with different gene expression profiles. The first, called germinal center B cell-like (GCB), develops from normal germinal center (GC) B-cells. The second, called Activated B cell-like (ABC), is thought to develop from plasmablasts, a population of B cells at a later stage of differentiation. In agreement with these cellular origins, the somatic hypermutation process (SHM), responsible for an aberrant accumulation of chromosomal translocations and somatic mutations at multiple oncogenes in these pathologies, remains ongoing in all GCB but in only rare ABC cases. Whether this ongoing SHM activity also participates in clinical evolutions is not known. To address this question, we have reconstructed the phylogenetic trees of 14 refractory/relapsed DLBCL cases by analysing the somatic mutations patterns at the IGH variable regions from different biopsies, obtained at diagnosis and during progressions. These lymphomas were classified as belonging to the GCB or ABC subtype by hierarchical clustering, using the expression of 18 genes that discriminate these two molecular subtypes. For each of the 30 biopsies we analysed, the clonotypic VDJ rearrangement was amplified by PCR, subcloned into the pGEM-T vector, and 10 individual colonies were sequenced. Phylogenetic trees rooted on the closest IGVH germline gene were reconstructed using the SeaView software and the PhyML algorithm. To avoid PCR artefact, only those mutations observed in at least two subclones were considered. In this series, hierarchical clustering identified 5 ABC and 5 GCB DLBCLs. The four remaining cases showed intermediate gene expression profiles. As expected, intraclonal variations (ICV) were found in all 5 GCB and in only 1 ABC tumor at diagnosis. Three lymphomas, all from the ABC subtype, did not show any evidence of clonal evolution, presenting homogeneous and identical SHM patterns at diagnosis and progression. For five other cases (4 GCB and 1 undefined DLBCL), the evolution trees revealed that the progressions have resulted from the selection of cells already present at diagnosis. Surprisingly, 3 of these 4 GCB cases showed ICV in the first but not in the second biopsy, suggesting that the SHM process was not anymore active in the selected population. Finally, for the six remaining cases (2 ABC, 1 GCB and 3 undefined DLBCLs), our data revealed that the tumoral populations at diagnosis and at relapse have evolved in parallel from less mutated precursors (Figure 1). In these lymphomas, the tumoral cells within the different biopsies shared the same CDR3 region, confirming their clonal origin. They also shared several somatic mutations, suggesting that their precursors were of GC or post-GC origin. Interestingly, 3 of these cases relapsed as lower grade follicular lymphomas (FL). One, from the GCB subtype, showed ICV both at diagnosis and at relapse. The two others, one ABC and one undefined case, showed ICV only at relapse, in FL, but not at diagnosis in DLBCL, again suggesting that the activity of the SHM process can be influenced by other factors than the ABC/GCB signature. In conclusion, our data show that progressions in DLBCL can develop from the dominant tumoral population or from the selection of minor subclones, maybe due to their inherent capacity to resist treatments. Alternatively, a significant percentage of clinical evolutions apparently develop from a pool of less mutated post-GC precursors, involved in an active diversification process driven by the SHM machinery. Disclosures: No relevant conflicts of interest to declare.

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Rituximab and dose-adjusted EPOCH (R-EPOCH) for treatment of aggressive B-cell non-Hodgkin lymphomas: A developing country perspective.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.e19503 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. e19503-e19503

Author(s):

Hasmukh Jain ◽

Tanuja Shet ◽

Epari Sridhar ◽

Hari Menon ◽

Uma Bhaskar Dangi ◽

...

Keyword(s):

B Cell ◽

Germinal Center ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Clinicopathological Features ◽

Hiv Positive ◽

Cd4 Counts ◽

Poor Risk ◽

Grade 3

e19503 Background: Biological heterogeneity of diffuse large B-cell lymphomas (DLBL) is reflected in variable outcomes with R-CHOP. Clinicopathological features like high IPI, underlying acquired immunodeficiency, non-germinal center phenotype, primary mediastinal B-cell histology and Bcl-2 positivity correlate with adverse biology. Effect of adverse biological attributes is also reflected in outcomes B-cell lymphoma intermediate between diffuse large cell B-cell and Burkitt’s lymphomas (BCLU) and Burkitt’s lymphoma in elderly and HIV-positive individuals with low CD4 counts. Given the promising efficacy of R- EPOCH in poor-risk DLBL we used it in therapy of aggressive B-cell NHL. Methods: From May 2010 to December 2012 patients with de-novo aggressive B-cell lymphomas with adverse clinicopathological features (DLBL with multiple extranodal sites, high IPI, primary mediastinal B-cell lymphoma, BCLU, Burkitt’s lymphoma in elderly or in HIV-positive individuals) were treated with R-EPOCH. Demography,stage, bulk of disease, IPI, CD4 counts, grade 3/4 toxicities and death were recorded. Responses were evaluated at mid and end of therapy. Overall and progression-free survivals were calculated. Immunohistochemistry was used to classify the germinal center and non-germinal center phenotype. Results: 33 patients (males-21, female-12) were treated with median of 6 cycles (range 3-6) of R-EPOCH. Histologicalsubtypes distribution was- DLBL-16, BL-8, BCLU-5, and PMBL-4. Sixty percent of DLBL were non-germinal center type and bcl-2 positive. Median age was 33 years (range 15-73 years). More than 2/3 of patients had stage 3/4 bulky disease with high IPI. Twelve patients were HIV positive with median CD4 count of 173/µL (range 43-359/µL). Complete responses were seen in 66.7% at interim evaluation and in 90% at the end of therapy. At a median follow up of 8 months, 1- year overall survival and progression- free survival were 72.5% and 90% respectively. Two patients died due to therapyand grade 3/4 toxicities were seen in 10% patients. Conclusions: R-EPOCH regimen is efficacious in patients with poor-risk aggressive B-cell NHL however longer follow-up is needed.

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