Molecular Dissection of Diffuse Large B-Cell Lymphoma

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. SCI-14-SCI-14
Author(s):  
Laura Pasqualucci

Diffuse large B-cell lymphoma (DLBCL), the most common form of human lymphoma, is an aggressive malignancy comprising multiple phenotypically and genetically distinct subtypes, approximately 40% of which are incurable. These tumors may arise de novo or from the transformation of more indolent lymphomas, as observed in 30-40% of follicular lymphoma (FL) and 5-12% of chronic lymphocytic leukemia cases. Over the last decade, the introduction of next-generation sequencing technologies combined with genome-wide copy number analysis has allowed a comprehensive definition of the genetic lesions that are associated with the pathogenesis of these malignancies, leading to the identification of several previously unappreciated targets1-3. These lesions, in turn, uncovered dysregulated cellular pathways that represent potential targets for improved diagnosis and therapy. Among the most common genetic alterations found in both de novo DLBCL and transformed FL (tFL) are those targeting histone/chromatin modifiers; in particular, loss-of-function mutations in the genes encoding for the H3K4 methyltransferase MLL2 and the acetyltransferases CREBBP/EP300, together with gain-of-function mutations of the EZH2 H3K27 methyltransferase are observed in over 50% of DLBCL and 90% of tFL patients, suggesting a major role for these enzymes in altering gene expression during malignant transformation. Interestingly, sequential analysis of tumor samples isolated at FL diagnosis and at evolution to DLBCL indicates that inactivating mutations of CREBBP and MLL2 represent early events acquired during the initial expansion of a common ancestral clone4. Disruption of epigenetic modifiers by genetic alterations may thus contribute to malignant transformation by shaping the epigenetic landscape of the cancer cell as well as by perturbing specific biological programs. In line with this hypothesis, we have shown that mutations of CREBBP/EP300 disrupt the balance between acetylation-mediated activation of the p53 tumor suppressor and inactivation of the BCL6 proto-oncogene5. The lecture will cover recent advances in our understanding of the genetic basis of this disease, with emphasis on the role of epigenetic regulators in normal germinal center development and lymphomagenesis, as revealed by in vitro and in vivo studies. References: 1. Pasqualucci L, Trifonov V, Fabbri G, et al. Analysis of the coding genome of diffuse large B-cell lymphoma. Nat Genet. 2011; 43: 830-837. 2. Morin RD, Mendez-Lago M, Mungall AJ, et al. Frequent mutation of histone-modifying genes in non-Hodgkin lymphoma. Nature. 2011; 476: 298-303. 3. Lohr JG, Stojanov P, Lawrence MS, et al. Discovery and prioritization of somatic mutations in diffuse large B-cell lymphoma (DLBCL) by whole-exome sequencing. Proc Natl Acad Sci U S A. 2012; 109: 3879-3884. 4. Pasqualucci L, Khiabanian H, Fangazio M, et al., Genetics of follicular lymphoma transformation. Cell Rep. 2014; 6:130-140. 5. Pasqualucci L, Dominguez-Sola D, Trifonov V, et al. Inactivating mutations of acetyltransferase genes in B-cell lymphoma. Nature. 2011; 471: 189-195. Disclosures No relevant conflicts of interest to declare.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2819-2819
Author(s):  
Matthew J. Maurer ◽  
Thomas E. Witzig ◽  
William R. Macon ◽  
Sergei I. Syrbu ◽  
James R. Cerhan ◽  
...  

Abstract Background: A significant percentage of DLBCL patients present with a composite histology, often seen as a node containing both follicular lymphoma and DLBCL or DLBCL in the node and discordant indolent lymphoma in the bone marrow. Literature from the pre-rituximab era suggests DLBCL patients with transformed lymphoma or composite histology have worse outcome than de novo DLBCL. Here we report on early events in a cohort of R-CHOP treated patients. Goal: To determine whether patients with composite lymphoma have an inferior event free survival (EFS) and overall survival (OS) compared to de novo diffuse large B-cell lymphoma when treated with R-CHOP. Methods: Newly diagnosed patients treated with an R-CHOP containing regimen were prospectively enrolled in our Lymphoma SPORE registry from 9/2002 through 6/2007. Pathology was centrally reviewed. All patients were followed for retreatment, disease progression, and death. Results: 401 DLBCL patients were enrolled; 14% (57/401) had a composite histology. 33 patients had DLBCL and another histology, predominantly follicular lymphoma (n=29), in the same node. 20 patients had a non-DLBCL histology in a distinct location from the DLBCL; this was primarily indolent lymphoma in the bone marrow (n=15). 4 patients had both. 19% (75/401) of patients died during follow-up and 30% (121/401) had an event (death due to any cause, progression, or retreatment). Median follow-up for living patients was 34 months (range, 5–73). Composite DLBCL patients had higher event-free (3 year EFS = 79%) and overall (3 year OS = 93%) survival then de novo DLBCL (3 year OS = 66%, 3 year EFS 79%), p=0.05 and p=0.005 respectively. These differences remained statistically significant after adjusting for the International Prognostic Index (IPI): EFS HR = 0.53, 95% CI: 0.29–0.97, p=0.02; OS HR=0.28, 95% CI: 0.10–0.76, p=0.002. OS and EFS for composite DLBCL more closely resembled R-CHOP treated grade III follicular lymphoma (A,B) from the same cohort (3 year EFS = 81%, 3 year OS = 93%). Improved outcome for composite DLBCL was consistent whether the additional histology was in the same node or distinct from the DLBCL. Conclusions: R-CHOP treated DLBCL patients with indolent discordant bone marrow involvement or other composite histology have improved early OS and EFS compared to de novo DLBCL. Further follow-up is needed to assess the long-term prognosis of composite DLBCL in the rituximab era. Histology N Age > 60 Stage III/IV LDH > ULN PS > 1 >2 EN Sites 3 YR EFS 3 YR OS * Denotes statistically significant difference at p=0.05 de novo DLBCL 344 58% 56% 56% 17% 22% 66% 78% Composite DLBCL 57 65% 77%* 34%* 18% 32% 79%* 93%* Figure Figure


Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 1284-1284
Author(s):  
Carol Y Ying ◽  
David Dominguez-Sola ◽  
Melissa Fabi ◽  
Ivo C Lorenz ◽  
Mukesh Bansal ◽  
...  

Abstract Abstract 1284 Diffuse large B-cell lymphoma (DLBCL) and Follicular Lymphoma (FL) are the most common forms of non-Hodgkin's lymphoma in the adult, accounting for approximately 75% of lymphoma diagnoses. Recent technological advances, including whole-genome DNA and RNA sequencing and gene copy-number analysis, have provided a comprehensive view of the genomic landscape of DLBCL, allowing new insights in the somatic genetic lesions that are associated with the pathogenesis of this malignancy. Among the genetic alterations that are recurrently found in DLBCL and FL, but remain of unclear functional significance, are the mutations involving the MEF2B gene. MEF2B is a member of the myocyte enhancer-binding factor 2 (MEF2) family of transcription factors whose activity is dependent on association with specific co-repressors (including CABIN1 and HDACs) and co-activators in response to multiple signaling pathways. Overall, ∼11% of DLBCL and ∼12% of FL cases reported carry mutations in MEF2B (Morin Nature 2011; Pasqualucci Nat Genet 2011; Lohr PNAS 2012). We showed that within the mature B-cell lineage, MEF2B expression is restricted to germinal center (GC) B-cells. The analysis of the B-cell interactome, a network of protein-protein and protein-DNA interactions generated by reverse-engineering a large dataset of B-cell phenotypes, showed that MEF2B was uniquely connected to BCL6, a proto-oncogene and well-characterized master regulator of the GC reaction. We demonstrated that MEF2B directly binds to the promoter of BCL6 and leads to its trans-activation in GC B-cells. Consistently, silencing of MEF2B in GC-derived lymphoma cell lines led to BCL6 down-regulation and impairment of cell cycle progression and proliferation, suggesting that normal and malignant GC cells are dependent on MEF2B expression. Approximately 80% of the DLBCL and FL mutated cases carry missense mutations clustered in the N-terminal conserved MADS-box and MEF2 functional domains, suggesting that they may have a relevant impact on MEF2B function. In a second group of cases (∼20%), mutations affect the C-terminal half of the MEF2B protein, and are mostly represented by frameshift and nonsense mutations, which truncate or modify the C-terminus of the protein. In order to functionally characterize these mutations, we first investigated whether DLBCL- and FL-associated MEF2B mutations affected the ability to regulate the transcription of BCL6. Using a reporter construct containing the native BCL6 promoter region responsive to MEF2B, we demonstrated that most of the N-terminal mutations cause increased transcriptional activity as tested on the BCL6 promoter. The analysis of the N-terminal MEF2B crystal structure, upon mapping the mutated residues, predicted that these mutations may interfere with the ability of MEF2B to heterodimerize with the CABIN1 co-repressor. Indeed, we showed that these MEF2B mutant proteins fail to bind CABIN1 and are resistant to its transrepressive activity. Conversely, C-terminal MEF2B mutations lead to truncated MEF2B proteins lacking the domains responsive to two independent post-transcriptional modifications, namely PKA-mediated phosphorylation and sumoylation. We showed that MEF2B is in fact phosphorylated by PKA and sumoylated in vivo, that both of these modifications lead to negative regulation of MEF2B transcriptional activity, and that lymphoma-associated C-terminal mutants fail to be negatively regulated by PKA-mediated phosphorylation and sumoylation. In summary, these results identify MEF2B as an upstream regulator of BCL6 and GC formation, which is required for lymphoma proliferation. Lymphoma-associated MEF2B mutations may contribute to lymphomagenesis, at least in part, by deregulating the expression of the BCL6 oncogene. Thus, targeting MEF2B may represent an alternative therapeutic approach to block BCL6 and cell proliferation in DLBCL and FL. Disclosures: No relevant conflicts of interest to declare.


Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 2666-2666
Author(s):  
Shin-ichiro Fujiwara ◽  
Kazuo Muroi ◽  
Yuji Hirata ◽  
Kazuya Sato ◽  
Tomohiro Matsuyama ◽  
...  

Abstract Abstract 2666 Background: CD25 (the alpha chain of the IL-2 receptor) has been shown to be expressed in a small subset of normal B cells and hairy cell leukemia cells; however, its expression in diffuse large B-cell lymphoma (DLBCL) has not been examined. The aim of this study was to clarify the clinical features of CD25+ DLBCL. Patients and Methods: Fifty-five patients with newly diagnosed CD25+ DLBCL who were admitted to our hospital between 1993 and 2011 were retrospectively evaluated. Lymph node or related tissue biopsy specimens from the patients were analyzed using FCM combined with single- and two-color staining. CD25 expression was defined as positivity of >20% of clonal B cells with CD19 or 20 expression >80% in a gated region. Results: There were 30 males and 25 females, with a median age of 65 years (range, 27–88 years). They showed aggressive clinical features as follows: 37 patients older than 60 (67%), 45 with elevated LDH (82%), 42 with soluble IL-2 receptor higher than 2,000 U/ml (79.4%), 33 with advanced-stage disease (stage III or IV, 60%), 28 with more than one extra nodal site (51%), 31 at high/high-intermediate risk according to the international prognostic index (IPI) score (56.4%). Chromosomal abnormalities were identified in 27 (79.4%) of 34 patients and frequently at chromosomes 3 (n = 13), 6, 7, 8, 14 (n = 9, respectively), and 1 (n = 7). CD25 expression showed a mean of 60.2% (range, 21.2–97.4%), and was particularly higher in patients aged more than 65 years and with CNS involvement (66.8 vs. 52.8%, p= 0.03 and 88.2 vs. 61.8%, P <0.01, respectively). In two-color FCM analysis, the percentages (mean ± SD) of CD19+CD25+ and CD20+CD25+ cells were 63.7 ± 25.5% (n=13) and 55 ± 28.1% (n=14), respectively. Compared to the patients with de novo CD25+ follicular lymphoma (CD25>20%, CD19/20>80%, n = 7), CD25 expression was significantly higher in CD25+ DLBCL (59.1%) than in CD25+ follicular lymphoma (34.5%). The complete remission (CR), 4-year progression free survival (PFS) and overall survival rates in the patients treated with R-CHOP (n = 35) were 81, 47, and 64%, respectively. When CD25 expression levels were compared between patients in CR and alive and those in non-CR and deceased, a trend was shown (58.1 vs. 74.4%, P = 0.055, and 57 vs. 71.8%, P = 0.058, respectively). Conclusion: CD25+ DLBCL may constitute a distinct subgroup with aggressive clinical features. Because R-CHOP is less effective for these patients, an alternative approach such as R-CHOP + anti-CD25 immunotherapy is needed. Disclosures: No relevant conflicts of interest to declare.


Hematology ◽  
2020 ◽  
Vol 2020 (1) ◽  
pp. 306-311
Author(s):  
Sonali Smith

Abstract Although the majority of indolent lymphomas (focusing on follicular lymphoma [FL]) have a prolonged waxing and waning course, a portion of patients experience histologic transformation (HT) to either diffuse large B-cell lymphoma or a higher-grade morphology, often with acquisition of MYC and BCL2 and/or BCL6 rearrangements (high-grade B-cell lymphoma–double-hit lymphoma/triple-hit lymphoma). The overall incidence of HT and transformed follicular lymphoma (tFL) may be declining, but outcomes remain inferior to those in simple indolent lymphoma progression. Recent data suggest that the majority of HT cases occur in higher-risk patients with FL, and they occur early after initial chemoimmunotherapy, comprising the majority of patients with progression of disease within 24 months. This latter point emphasizes the need for a sufficient biopsy at relapse in FL. Treatment options depend on the prior therapy for the indolent component as well as the histology at relapse, but they generally follow several principles discussed in this article. Anthracycline-naïve patients have the best outcomes if there is HT, and responses to R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, prednisone) are similar to those of patients with de novo diffuse large B-cell lymphoma. Patients with anthracycline exposure prior to transformation have the best outcomes with salvage chemotherapy and a consolidative autologous stem cell transplant. However, a major challenge is the management of patients with tFL who experience relapse early after bendamustine-based treatment, in whom the role of consolidative transplant after anthracycline-based treatment is unclear. In the past several years, cellular therapy has emerged as an important tool for some but not all patients with tFL. This review focuses on the nuances of managing tFL.


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