Prognostic Significance of MYC Single, Double, Triple Hit and MYC-Translocation Partner Status in Diffuse Large B-Cell Lymphoma - a Study By the Lunenburg Lymphoma Biomarker Consortium (LLBC)

Abstract Introduction MYC-rearrangement (MYC-R) occurs in approximately 10-15% of diffuse large B-cell lymphomas (DLBCL) and several studies suggest an inferior progression free (PFS) as well as overall survival (OS) compared to DLBCL without MYC-R. However, the prognostic significance of MYC single-hit (MYC-SH), MYC double/triple-hit (MYC and BCL2 and/or BCL6 translocation; MYC-DH/TH) in the context of the MYC translocation partner (MYC-IG versus MYC-non-IG) is less clear due to relatively small sample sizes in prior studies. The Lunenburg Lymphoma Biomarker Consortium (LLBC) set out to address these questions in a large number of DLBCL treated uniformly with rituximab (R)-CHOP- or R-CHOP-like chemotherapy. Methods Relevant clinical data (IPI factors, PFS, OS) from patients with aggressive B-cell lymphomas with confirmed DLBCL morphology derived from registry cohorts (Canada: British Columbia Cancer, UK: Leeds/HMRN, Barts, USA: Stanford) and prospective clinical trials (Germany: RICOVER, MegaCHOEP, France: LNH01-5B and LNH03-B, HOVON: HO 46 and 84) were pooled in the LLBC database (n=5118). Interphase FISH analysis (mostly on tissue microarrays) was used to determine the MYC-, BCL2- and BCL6-rearrangement status. All DLBCL with available tissue underwent break-apart testing for MYC (Vysis, LSI, Abbott). Identified MYC-R cases were subjected to FISH break-apart testing for the BCL2- and BCL6 loci as well as to MYC/IGH fusion testing and, if negative, to MYC/IGK and MYC/IGL testing to allow designation of the final MYC-R status (MYC-IG versus MYC-non-IG). Information on the cell of origin (COO) was generated using immunohistochemistry (Hans classifier) and/or gene expression based methods. Survival probabilities were estimated using the Kaplan Meier method and survival curves were compared with the log-rank test. The effects of variables of interest were estimated using univariate and multivariate Cox models stratified on the variable 'cohort' and 'trial'. Results 2380 DLBCL patients with full data on PFS, OS, IPI factors and MYC FISH results were available for analysis. 263 DLBCL cases (11%) had MYC-R which was associated with inferior PFS and OS compared to DLBCL patients without MYC-R (59% versus 72% OS at five years; log-rank <0.001; 56% versus 64% PFS at five years; log-rank <0.002). This effect was observed in both COO subgroups (GCB and non GCB). Out of 163 MYC-R patients with complete FISH data, 40 were MYC-SH with an IG partner (24.5%) and 17 (10.5%) with a non-IG partner. 53 patients had a MYC DH/TH constellation with an IG partner (32.5%) and 53 patients (32.5%) with a non-IG partner. The MYC-DH/TH group with an IG partner had the worst OS at 24 months: (50.9% compared to 76.4% for the remaining MYC-R DLBCL patients and to 82.4% for patients without MYC-R) (fig1). Similar results were obtained for PFS. In the MYC-DH group, there were no differences in OS and PFS between MYC/BCL2 and MYC/BCL6 'double hits'. Multivariate Cox models adjusting for the IPI and including a time-dependent effect showed, that the impact on outcomes was mainly seen in the first 24 months post therapy (hazard ratio (HR) of 2 [1.59-2.53] for MYC-R DLBCL patients compared to patients without MYC-R and a HR of 3.12 [2.09-4.64] for MYC-DH/TH tumors in which MYC is translocated to IG). In contrast, in MYC-R patients including MYC-SH and MYC-DH/TH without an IG partner, the HR was lower at 1.5 [1.01-2.24]. Conclusion This study by the LLBC in a very large cohort of DLBCL patients treated with R-CHOP or R-CHOP-like therapy confirms previous reports on the negative prognostic impact of an underlying MYC-translocation for both PFS and OS. This impact is predominantly observed in the first two years post therapy. Further, the large sample size extends previous observations that the partner gene of MYC (IG versus non-IG) also has prognostic impact. MYC-DH/TH DLBCL with an IG partner gene have the worst OS and PFS, while impact is moderate in other constellations (MYC-SH and MYC-DH/TH with a non-IG partner). Additionally, no differences between MYC/BCL2 and MYC/BCL6 'double hits' are seen in PFS and OS. Our results suggest that along with MYC testing in routine clinical practice, identification of the MYC partner gene (IG versus non-IG) is also warranted to identify further DLBCL subsets with poor outcomes which may have implications in the design and interpretation of future clinical trials. Figure 1. Figure 1. Disclosures Sehn: Merck: Consultancy, Honoraria; TG Therapeutics: Consultancy, Honoraria; Roche/Genentech: Consultancy, Honoraria; Amgen: Consultancy, Honoraria; Seattle Genetics: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; Morphosys: Consultancy, Honoraria; Karyopharm: Consultancy, Honoraria; Lundbeck: Consultancy, Honoraria; Janssen: Consultancy, Honoraria; Abbvie: Consultancy, Honoraria.

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Prognostic Significance of MYC Rearrangement and Translocation Partner in Diffuse Large B-Cell Lymphoma: A Study by the Lunenburg Lymphoma Biomarker Consortium

Journal of Clinical Oncology ◽

10.1200/jco.19.00743 ◽

2019 ◽

Vol 37 (35) ◽

pp. 3359-3368 ◽

Cited By ~ 26

Author(s):

Andreas Rosenwald ◽

Susanne Bens ◽

Ranjana Advani ◽

Sharon Barrans ◽

Christiane Copie-Bergman ◽

...

Keyword(s):

B Cell ◽

Prognostic Significance ◽

B Cell Lymphoma ◽

Study Cohort ◽

Prognostic Impact ◽

Patient Registries ◽

Biopsy Material ◽

Partner Gene ◽

The Impact ◽

Large B Cell

PURPOSE MYC rearrangement ( MYC-R) occurs in approximately 10% of diffuse large B-cell lymphomas (DLBCLs) and has been associated with poor prognosis in many studies. The impact of MYC-R on prognosis may be influenced by the MYC partner gene (immunoglobulin [IG] or a non-IG gene). We evaluated a large cohort of patients through the Lunenburg Lymphoma Biomarker Consortium to validate the prognostic significance of MYC-R (single-, double-, and triple-hit status) in DLBCL within the context of the MYC partner gene. METHODS The study cohort included patients with histologically confirmed DLBCL morphology derived from large prospective trials and patient registries in Europe and North America who were uniformly treated with rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone therapy or the like. Fluorescence in situ hybridization for the MYC, BCL2, BCL6, and IG heavy and light chain loci was used, and results were correlated with clinical outcomes. RESULTS A total of 5,117 patients were identified of whom 2,383 (47%) had biopsy material available to assess for MYC-R. MYC-R was present in 264 (11%) of 2,383 patients and was associated with a significantly shorter progression-free and overall survival, with a strong time-dependent effect within the first 24 months after diagnosis. The adverse prognostic impact of MYC-R was only evident in patients with a concurrent rearrangement of BCL2 and/or BCL6 and an IG partner (hazard ratio, 2.4; 95% CI, 1.6 to 3.6; P < .001). CONCLUSION The negative prognostic impact of MYC-R in DLBCL is largely observed in patients with MYC double hit/triple-hit disease in which MYC is translocated to an IG partner, and this effect is restricted to the first 2 years after diagnosis. Our results suggest that diagnostic strategies should be adopted to identify this high-risk cohort, and risk-adjusted therapeutic approaches should be refined further.

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Overexpression of BCL-2 Does Not Inhibit Autophagy in Human Follicular and Diffuse Large B-Cell Lymphomas

Blood ◽

10.1182/blood.v124.21.3007.3007 ◽

2014 ◽

Vol 124 (21) ◽

pp. 3007-3007

Author(s):

Aine McCarthy ◽

Andrew James Clear ◽

Jacek Marzec ◽

Rita Coutinho ◽

Robert D. Petty ◽

...

Keyword(s):

B Cells ◽

Protein Expression ◽

B Cell ◽

Prognostic Significance ◽

B Cell Lymphoma ◽

Beclin 1 ◽

Pcr Array ◽

B Cell Lymphomas ◽

Autophagy Activity ◽

Large B Cell

Abstract Background: Human follicular lymphoma (FL) and diffuse large B-cell lymphoma (DLBCL) are the most common forms of indolent and aggressive NHL, respectively. The t(14;18) translocation characterizes approximately 85% of FL and 20% of DLBCL and results in constitutive overexpression of the anti-apoptotic protein BCL-2. It was previously reported that BCL-2 plays dual roles in preventing apoptosis and autophagy. Autophagy is a physical and pathological process whereby cells sequester portions of cytoplasm including organelles to form autophagosomes where they are degraded and recycled. Growing evidence demonstrates that autophagy plays important roles in tumorigenesis, tumor progression, and resistance to chemotherapy. Aims: The autophagy status in human B-cell lymphomas is unknown. We hypothesized that overexpression of BCL-2 could change autophagy status and aimed to determined expression of autophagy-related genes and proteins in FL and DLBCL primary samples by PCR array and tissue microarray. We aimed to evaluate whether expression of the autophagy-related proteins p62, Beclin-1 and LC3 individually and in combination with BCL-2 protein expression could risk-stratify FL and DLBCL patients at diagnosis. Patients and methods: Using PCR array, the autophagy-related gene expression profiles were determined in purified and unpurified reactive and malignant human lymph node tissue biopsies. Diagnostic tissues from FL (n=117) and DLBCL (n=109) patients were microarrayed and autophagy protein expression was evaluated using immunohistochemistry. Univariate and multivariate analyses on both continuous and categorical variables were conducted to measure overall survival (OS), disease specific survival (DSS), and progression-free survival (PFS). Results: Seven autophagy machinery genes were up-regulated in purified FL B-cells, namely ATG9A, ATG16L1, MAP1LC3A, GABARAPL1, ULK1, LAMP1 and HDAC6 compared with reactive B-cells. Two autophagy machinery genes, MAP1LC3A and DRAM1, were up-regulated in DLBCL B-cells. In unpurified tissue biopsies, 20 of 46 genes in FL and 2 of 5 genes in DLBCL with increased expression were autophagy machinery genes. CTSD (cathepsin D) and TGM2 (transglutaminase 2) genes and proteins were mainly up-regulated in DLBCL tumor-infiltrating macrophages. These results demonstrate that FL and DLBCL showed increased expression of autophagy-related genes, regardless of the heterogeneity of these diseases. p62, a selective autophagy substrate; LC3, an autophagosome membrane protein; and Beclin-1, an essential autophagy effector, are often used to evaluate autophagy activity in the cell. 91% FL samples were BCL-2 positive but significantly decreased expression of p62, LC3 and Beclin-1 in FL samples was observed in both intra-follicular and non-malignant inter-follicular areas. This suggests that increased basal autophagy activity in both malignant FL cells and surrounding tumor infiltrating cells, indicating that BCL-2 does not inhibit basal autophagy activity. DLBCL samples displayed heterogeneous expression patterns of BCL-2, p62, LC3 and Beclin-1. We found that decreased p62 expression confers worse OS (continuous P=0.015; and categorical P=0.003), DSS (continuous P=0.037; categorical P=0.014) and PFS (categorical P=0.002) in DLBCL patients. Decreased expression of Beclin-1 was also confers poor prognosis in both FL and DLBCL as conducted by categorical analysis, OS (DLBCL, P=0.015; FL, P=0.004), DSS (FL, P=0.006), and PFS (DLBCL, P=0.029). p62 retains prognostic significance after adjustment for the International Prognostic Index (IPI) score and levels of BCL-2, Beclin-1 and LC3 in multivariate analysis. Beclin-1 retains its prognostic significance in FL after adjusting for FLIPI scores. Low p62 plus high BCL-2 expression in DLBCL confers the worst OS (P<0.0001) and DSS (P=0.001) compared with other combinations. Conclusions: These results demonstrate that FL has increased basal autophagy activity, while it varies in DLBCL. p62 is a novel, independent prognostic biomarker for DLBCL but not for FL. Combining p62 with BCL-2 provides a more robust and reliable method to risk-stratify DLBCL patients at diagnosis. Importantly, we report for the first time that overexpression of BCL-2 in human NHL does not inhibit basal autophagy activity. We propose that increased autophagy activity could be a therapeutic target for treatment of NHL. Disclosures Gribben: Celgene: Research Funding; Pharmacyclics: Honoraria; Roche: Honoraria.

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Cell-of-origin classification using the Hans and Lymph2Cx algorithms in primary cutaneous large B-cell lymphomas

Virchows Archiv ◽

10.1007/s00428-021-03265-5 ◽

2022 ◽

Author(s):

Anne M. R. Schrader ◽

Ruben A. L. de Groen ◽

Rein Willemze ◽

Patty M. Jansen ◽

Koen D. Quint ◽

...

Keyword(s):

B Cell ◽

Cell Lymphoma ◽

Prognostic Significance ◽

B Cell Lymphoma ◽

Large Cell ◽

Cell Of Origin ◽

B Cell Lymphomas ◽

Large B Cell Lymphoma ◽

And Behavior ◽

Large B Cell

Abstract Primary cutaneous diffuse large B-cell lymphoma, leg type (PCDLBCL-LT) and primary cutaneous follicle center lymphoma with a diffuse population of large cells (PCFCL-LC) are both primary cutaneous B-cell lymphomas with large-cell morphology (CLBCL) but with different clinical characteristics and behavior. In systemic diffuse large B-cell lymphoma, not otherwise specified (DLBCL-NOS), gene-expression profiling (GEP) revealed two molecular subgroups based on their cell-of-origin (COO) with prognostic significance: the germinal center B-cell-like (GCB) subtype and the activated B-cell-like (ABC) subtype. This study investigated whether COO classification is a useful tool for classification of CLBCL. For this retrospective study, 51 patients with PCDLBCL-LT and 15 patients with PCFCL-LC were analyzed for their COO according to the immunohistochemistry-based Hans algorithm and the NanoString GEP-based Lymph2Cx algorithm. In PCFCL-LC, all cases (100%) classified as GCB by both Hans and Lymph2Cx. In contrast, COO classification in PCDLBCL-LT was heterogeneous. Using Hans, 75% of the PCDLBCL-LT patients classified as non-GCB and 25% as GCB, while Lymph2Cx classified only 18% as ABC, 43% as unclassified/intermediate, and 39% as GCB. These COO subgroups did not differ in the expression of BCL2 and IgM, mutations in MYD88 and/or CD79B, loss of CDKN2A, or survival. In conclusion, PCFCL-LC uniformly classified as GCB, while PCDLBCL-LT classified along the COO spectrum of DLBCL-NOS using the Hans and Lymph2Cx algorithms. In contrast to DLBCL-NOS, the clinical relevance of COO classification in CLBCL using these algorithms has limitations and cannot be used as an alternative for the current multiparameter approach in differentiation of PCDLBCL-LT and PCFCL-LC.

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Clinical Features and Prognostic Significance of NOTCH1 Mutations in Diffuse Large B-Cell Lymphoma

Frontiers in Oncology ◽

10.3389/fonc.2021.746577 ◽

2021 ◽

Vol 11 ◽

Author(s):

Zhongqi Li ◽

Fang Yu ◽

Wenle Ye ◽

Liping Mao ◽

Jiansong Huang ◽

...

Keyword(s):

B Cell ◽

Clinical Features ◽

Cell Lymphoma ◽

International Prognostic Index ◽

Prognostic Significance ◽

Prognostic Index ◽

B Cell Lymphoma ◽

Prognostic Impact ◽

Large B Cell Lymphoma ◽

Large B Cell

Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous group of large lymphoid B cell malignancy with distinct clinical and genetic features. Recently, NOTCH1 mutations were identified in DLBCL cases by Next-generation sequencing (NGS), but the clinical features and prognostic impact were not systematically studied. Here, NOTCH1 genes in 161 DLBCL samples were sequenced by NGS. The prognostic value of NOTCH1 mutations was assessed in the context of clinical and laboratory factors, such as international prognostic index (IPI), cell-of-origin classification, double expression of BCL2 and c-MYC. The combined data from three Western cohorts were used to validate these results. As a result, NOTCH1 mutations were found in 17(10.6%) patients, and three patients had a hotspot mutation of c.7541_7542delCT. The presence of NOTCH1 mutations was significantly associated with poor complete response and progression free survival(PFS), which was independent of established clinical and laboratory parameters. In addition, 30 (1.92%) of 1562 patients treated with R-CHOP regimen in those combined Western cohorts had NOTCH1 mutations. Meta-analysis of the Western cohorts confirmed that NOTCH1 mutations were also associated with poor PFS and OS. In conclusion, DLBCL patients with the NOTCH1 mutations have worse PFS and OS, and the NOTCH1 mutations can be used as an independent predictor for patients with DLBCL.

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Prognostic Impact of Tumor Infiltrating FOXP3 Positive Regulatory T Cells in Diffuse Large B-Cell Lymphoma at Diagnosis.

Blood ◽

10.1182/blood.v110.11.1565.1565 ◽

2007 ◽

Vol 110 (11) ◽

pp. 1565-1565

Author(s):

Jae-Yong Kwak ◽

Na-Ri Lee ◽

Eun-Kee Song ◽

Kyu Yun Jang ◽

Ha Na Choi ◽

...

Keyword(s):

T Cells ◽

Regulatory T Cells ◽

B Cell ◽

Cell Lymphoma ◽

International Prognostic Index ◽

Prognostic Significance ◽

B Cell Lymphoma ◽

Prognostic Impact ◽

Large B Cell Lymphoma ◽

Large B Cell

Abstract Tumor infiltrating immune cells play an important role in host immune reactions against diffuse large B-cell lymphoma (DLBCL). In the present study, we have identified a subset of tumor infiltrating FOXP3-positive regulatory T cells (Tregs) in the initial DLBCL biopsy specimens and evaluated its prognostic significance. Ninety-six patients with DLBCL were retrospectively evaluated. Expression pattern of FOXP3 protein was examined using standard immunohistochemistry in paraffin-embedded tissue samples. The median overall survival (OS) was 28 months. Compared to the others, the patients with higher percentage of FOXP3-positive Tregs in the initial tumor biopsy, showed a significantly longer OS (p=0.003). When prognostic factors were evaluated in a multivariate model, the international prognostic index and the percentage of infiltrating FOXP3-positive Tregs in the initial biopsy were shown to be independent predictors of OS. In conclusion, the presence of increased percentage of FOXP3-positive Tregs in DLBCL predicts a better prognosis. Figure Figure

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Rearrangement of MYC Is Associated With Poor Prognosis in Patients With Diffuse Large B-Cell Lymphoma Treated in the Era of Rituximab

Journal of Clinical Oncology ◽

10.1200/jco.2009.26.3947 ◽

2010 ◽

Vol 28 (20) ◽

pp. 3360-3365 ◽

Cited By ~ 380

Author(s):

Sharon Barrans ◽

Simon Crouch ◽

Alex Smith ◽

Kathryn Turner ◽

Roger Owen ◽

...

Keyword(s):

Clinical Outcome ◽

B Cell ◽

Poor Prognosis ◽

International Prognostic Index ◽

Prognostic Significance ◽

Prognostic Index ◽

B Cell Lymphoma ◽

Prognostic Impact ◽

Large B Cell

Purpose Rearrangement of MYC occurs in a proportion of diffuse large B-cell lymphomas (DLBCL), where they may be associated with an adverse clinical outcome. The aim of this study was to determine the frequency of MYC translocations in DLBCL and their prognostic impact in the era of cyclophosphamide, doxorubicin, vincristine, and prednisone plus rituximab (CHOP-R) therapy. Patients and Methods Three hundred three patients with previously untreated DLBCL, with no evidence of underlying follicular lymphoma, were investigated using immunohistochemistry and interphase fluorescent in situ hybridization for MYC, BCL6, and t(14;18)/BCL2 rearrangements. All patients (median age, 71.1 years; range, 23 to 96 years) were treated when CHOP-R was standard therapy for DLBCL and observed for a maximum of 4 years. Overall survival (OS) at 3 years was 49% (95% CI, 42% to 56%). Results MYC rearrangements were demonstrated in 35 (14%) of 245 biopsies with data available. Of these, 26 (74%) also had a t(14;18), 10 (26%) were BCL6 and MYC rearranged, and seven had all three abnormalities. Only age, International Prognostic Index, and MYC rearrangement retained prognostic significance in the final model. OS was significantly worse for patients with rearrangement of MYC (survival probability at 2 years = 0.35 in v 0.61 in the nonrearranged group). Conclusion The presence of a MYC rearrangement is a strongly adverse prognostic factor in CHOP-R–treated patients and can be used in combination with patients' age and IPI to accurately predict clinical outcome. In DLBCL, rearrangement of MYC is rarely found as the sole genetic abnormality and the poor prognosis of these patients is likely to reflect a synergistic effect alongside deregulation of BCL6 or BCL2.

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Prognostic Significance of BCL6 Translocation in Diffuse Large B-Cell Lymphoma Patients Treated with R-CHOP

Blood ◽

10.1182/blood.v112.11.3782.3782 ◽

2008 ◽

Vol 112 (11) ◽

pp. 3782-3782

Author(s):

Jesse Shustik ◽

Christian Steidl ◽

Laurie Sehn ◽

Randy D Gascoyne

Keyword(s):

Overall Survival ◽

B Cell ◽

Prognostic Value ◽

Cell Lymphoma ◽

Prognostic Significance ◽

B Cell Lymphoma ◽

Immunohistochemical Analysis ◽

Prognostic Impact ◽

Large B Cell Lymphoma ◽

Large B Cell

Abstract Translocation of the BCL6 oncogene is the most commonly-detected chromosomal abnormality in diffuse large B-cell lymphoma (DLBCL), but its prognostic significance remains uncertain. BCL6 translocation has been associated with variable outcomes in prior clinical series, and its clinical impact has not been reevaluated since the introduction of rituximab (R). The objective of this study was to determine the prognostic value of BCL6 translocation in DLBCL patients treated with both CHOP and R-CHOP immunochemotherapy. Methods We included 163 DLBCL patients treated with CHOP (n=72) or R-CHOP (n=73) at the British Columbia Cancer Agency between the years 1999–2002. Formalin-fixed paraffin-embedded diagnostic lymph node specimens were analyzed for BCL6 translocation on tissue microarray (TMA) by fluorescence in situ hybridization (FISH), using commercially available dual-color break-apart probes (Abbott Molecular, IL, USA). Immunohistochemical analysis was performed for BCL6, CD10, and MUM1 expression, and cases were categorized as germinal centre B-cell-like (GCB) or non-GCB using a standard algorithm (Hans, Blood 2004). Results BCL6 translocation status was evaluable in 145 cases, and was positive (BCL6+) in 26% (37/145). Median age (65 vs. 59 years, p=0.03) and overall IPI score (p=0.03) were higher in the BCL6+ than in the BCL6− subgroup. Presence of BCL6 translocation correlated with non-GCB phenotype by immunohistochemical analysis (p=0.03), and showed no significant association with BCL6 protein expression (p=0.32). Median follow-up duration was 4.8 years (range, 0.1–8.4). BCL6 translocation was associated with a trend toward decreased overall survival (OS) among patients treated with R-CHOP (5-year OS 46.7% vs. 68%, p=0.098), though not among CHOP-treated patients (5-year OS 36.4% vs.49.5%, p=0.393). Kaplan-Meier analysis limited to the BCL6+ subgroup showed no overall survival difference between patients treated with CHOP and R-CHOP (p=0.67). Conclusions BCL6 translocations are found in approximately 25% of patients with de novo DLBCL. Presence of BCL6 translocation appears to predict unfavorable outcome in DLBCL patients treated with R-CHOP. However, the adverse prognostic impact of BCL6 translocation is confounded by an association with high IPI score and with non-GCB phenotype. Prospective analysis within large clinical trials would help to determine the independent prognostic value of this marker in the rituximab era.

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