Tumor-collagen digital proximity signature to indicate prognosis in diffuse large B-cell lymphoma.

e19073 Background: Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous tumor that originates from normal B-cells. Despite the use of combination chemotherapy, around 40% of DLBCL patients die (de Jonge, et al. European Journal of Cancer, 2016). Limited studies have investigated the role of collagen in the acellular tumor microenvironment. In this study, we present a novel digital signature of the proximity of tumor cells and collagen-VI (COL6) that can predict overall survival (OS) in DLBCL patients. To the best of our knowledge, this is the first study of its kind to employ automated image analysis. Methods: The proposed digital proximity signature (DPS) aggregates summary-level statistics from the entire whole slide image (WSI) and serves as a marker of regions, categorizing weak, moderate, significant, and strong tumor-collagen proximity and can be described as a surrogate for signaling. To accomplish this, we developed a novel artificial intelligence (AI) based multi-task model for simultaneous detection and classification of tumor cells and another bespoke method for automatically identifying COL6 fiber. The tumor-collagen proximity analysis was then performed by aggregating tumor cell statistics within the vicinity of COL6 fibres. Finally, the prognostic significance of DPS for OS in DLBCL was investigated with Kaplan-Meier analysis, stratifying patients into two groups based on the median of the DPS values. Results: We took WSIs of DLBCL tissue slides for 32 cases immunohistochemically stained with COL6 and Hematoxylin counterstain. The AI model for tumor cell identification achieved a high F1-score of 0.84, outperforming recent single-task learning models. Our results show that strong proximity of COL6 and tumor cells is linked to better OS in DLBCL patients ( p = 0.03). Conclusions: Our novel digitally computed COL6-tumor proximity signature shows prognostic significance for overall survival on a pilot dataset of 32 DLBCL patients. We are further validating the utility of this novel signature as a prognostic biomarker in larger cohorts of DLBCL patients.

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Prognostic significance of the aggregative perivascular growth pattern of tumor cells in primary central nervous system diffuse large B-cell lymphoma

Neuro-Oncology ◽

10.1093/neuonc/not012 ◽

2013 ◽

Vol 15 (6) ◽

pp. 727-734 ◽

Cited By ~ 16

Author(s):

Miaoxia He ◽

Changjing Zuo ◽

Jianjun Wang ◽

Jianmin Liu ◽

Binghua Jiao ◽

...

Keyword(s):

Central Nervous System ◽

Nervous System ◽

B Cell ◽

Tumor Cells ◽

Growth Pattern ◽

Cell Lymphoma ◽

Prognostic Significance ◽

B Cell Lymphoma ◽

Large B Cell Lymphoma ◽

Large B Cell

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Effect of addition of rituximab to CHOP on survival of patients in both the GCB and non-GCB subgroups of diffuse large B-cell lymphoma

Journal of Clinical Oncology ◽

10.1200/jco.2007.25.18_suppl.8040 ◽

2007 ◽

Vol 25 (18_suppl) ◽

pp. 8040-8040 ◽

Cited By ~ 1

Author(s):

K. Fu ◽

K. D. Perry ◽

L. M. Smith ◽

C. P. Hans ◽

T. C. Greiner ◽

...

Keyword(s):

Overall Survival ◽

B Cell ◽

De Novo ◽

Cell Lymphoma ◽

Prognostic Significance ◽

B Cell Lymphoma ◽

Rank Test ◽

Cell Of Origin ◽

Large B Cell Lymphoma ◽

Large B Cell

8040 Background: Diffuse large B-cell lymphoma (DLBCL) includes at least two prognostically important subgroups, i.e. germinal center B-cell (GCB) and activated B-cell (ABC) DLBCL, which were initially characterized by gene expression profiling and subsequently validated by immunostaining. Bcl-2 has also been identified as a prognostic indicator in the ABC subgroup. However, with the addition of rituximab (R) to standard chemotherapy, the prognostic significance of this subclassification of DLBCL is unclear. Methods: We studied 119 cases of de novo DLBCL including 70 cases treated with R-CHOP and 49 cases treated with CHOP. The cases were assigned to either the GCB or non-GCB subgroups using the methodology described by Hans et al (Blood 2004; 103:275). Characteristics of the patients were compared using the Chi-square test. Overall survival (OS) and event-free survival (EFS) were estimated using the Kaplan Meier method and compared with the log-rank test. Results: The median age of the 119 patients was 67 years, ranging from 20 to 90 years, and there were 62 males and 57 females. The clinical characteristics of patients treated with CHOP versus R-CHOP, including the IPI, were comparable. R-CHOP was more effective than CHOP with improved 5-year EFS (63% vs 41%, p=0.013) and OS (78% vs 47%, p<0.001). In both patient groups treated with R-CHOP or CHOP, the GCB subgroup had a significantly better 5-year EFS and OS compared to the non-GCB subgroup (OS: 91% vs 64% for R-CHOP, p=0.0073; 67% vs 31% for CHOP, p=0.034, respectively). Additionally, both the GCB and non-GCB subgroups treated with R-CHOP had a significantly improved OS compared to their respective subgroups receiving CHOP alone (GCB, p=0.015; non-GCB, p=0.019). Bcl-2 expression was not a significant predictor in either the GCB or non-GCB subgroups treated with R-CHOP (OS, GCB: p=0.32; non-GCB: p=0.43). Conclusions: In this retrospective study, we demonstrate that subclassification based on the cell of origin continues to have prognostic significance in patients with DLBCL treated with R-CHOP. Addition of rituximab to CHOP improves the overall survival of patients with DLBCL in both the GCB and non-GCB subgroups. No significant financial relationships to disclose.

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5-Hydroxymethylcytosines of Circulating Cell-Free DNA and Prognosis in Diffuse Large B-Cell Lymphoma

Blood ◽

10.1182/blood-2018-99-114375 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 2985-2985

Author(s):

Brian C.-H. Chiu ◽

Zhou Zhang ◽

Qiancheng You ◽

Elizabeth Stepniak ◽

Paige Bracci ◽

...

Keyword(s):

Overall Survival ◽

B Cell ◽

Cell Lymphoma ◽

Prognostic Significance ◽

B Cell Lymphoma ◽

Cell Free Dna ◽

Free Dna ◽

Large B Cell Lymphoma ◽

Large B Cell ◽

Circulating Cell Free Dna

Abstract Background: Elevated levels of circulating cell-free DNA (cfDNA) have been associated with poor prognosis and relapse in patients with diffuse large B-cell lymphoma (DLBCL). However, the tumor-specific molecular targets in cfDNA that have prognostic significance remain unclear. We investigated the association between 5-hydroxymethylcytosine (5hmC), a mark of active demethylation and gene activation, in cfDNA from plasma and prognosis in DLBCL. Methods: We used the 5hmC-Seal, a highly sensitive chemical labeling technique integrated with next-generation sequencing (NGS), to profile 5hmC in plasma cfDNA samples from Caucasian patients at the University of Chicago who were newly diagnosed with DLBCL (n=43) or follicular lymphoma (FL, n=28), the two most common histological subtypes of non-Hodgkin lymphoma (NHL), in 2011-13. Baseline clinical, laboratory, and vital status data were abstracted from medical records. Patients were followed through December 31, 2017 and those who relapsed after completion of treatment, lost to follow-up, or died were censored. We profiled 5hmC with 1-2 ng of cfDNA extracted from ~2 ml of plasma for library construction and the NGS. We obtained ~25 million reads per sample, providing a depth of coverage ~600X in terms of gene bodies. We normalized read counts and identified differential 5hmC markers using DESeq2. Cox proportional hazards model were used to estimate the association between 5hmC markers and overall survival. Results: We found that in cfDNA from DLBCL patients, 5hmC markers were enriched within gene bodies and depleted in CpG islands. The cfDNA-based 5hmC profiles at diagnosis differed between DLBCL and FL, and in DLBCL, the 5hmC profiles differed between Ann Arbor stage (stage 1/2 vs stage 3/4), lactate dehydrogenase (LDH) level (normal vs elevated), and cell-of-origin (germinal center B-like and activated B-cell-like DLBCL). In addition, genome-wide 5hmC distribution patterns in cfDNA samples are highly correlated with those found in cfDNA-paired tumor tissues, supporting the tumor relevance of cfDNA in a patient. Next, we evaluated the prognostic significance of cfDNA-based 5hmC in DLBCL using a two-step approach. In the discovery phase (7 DLBCL patients with relapse and 12 age- and sex-matched patients without relapse within two years following treatment), a substantial number of 5hmC markers were associated with relapse (449 gene bodies at 5% false discovery rate [FDR]). These relapse-associated 5hmC signatures showed high sensitivity, specificity, and overall accuracy (area under curve [AUC]=0.91) in predicting relapse in the independent validation set (relapse=5, no relapse=13). Finally, we identified a panel of 128 5hmC markers (fold change >20% and p-value <0.05) that were associated with 4-year overall survival. Conclusion: These findings suggest that 5hmC signatures in cfDNA at the time of DLBCL diagnosis correlate with standard clinical prognostic indices and hold promise as non-invasive markers for prognosis and survival. Disclosures Smith: BMS: Consultancy; Portola: Honoraria.

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BCL2 Expression Is a Prognostic Marker for the Activated B-Cell–Like Type of Diffuse Large B-Cell Lymphoma

Journal of Clinical Oncology ◽

10.1200/jco.2005.03.4264 ◽

2006 ◽

Vol 24 (6) ◽

pp. 961-968 ◽

Cited By ~ 192

Author(s):

Javeed Iqbal ◽

Vishala T. Neppalli ◽

George Wright ◽

Bhavana J. Dave ◽

Douglas E. Horsman ◽

...

Keyword(s):

Overall Survival ◽

B Cell ◽

Cell Lymphoma ◽

Mrna Level ◽

Prognostic Significance ◽

B Cell Lymphoma ◽

Large B Cell Lymphoma ◽

The Difference ◽

Large B Cell ◽

Bcl2 Expression

Background The role of BCL2 as a predictor of survival in diffuse large B-cell lymphoma (DLBCL) is controversial. DLBCL is heterogeneous, and the expression of BCL2 is variable within the two major subgroups of DLBCL, germinal center B-cell–like (GCB) and activated B-cell–like (ABC) DLBCL, as well as primary mediastinal DLBCL. Patients and Methods In this study, we investigated the correlation of BCL2 expression with survival in the two major subgroups of DLBCL, as well as the mechanisms of BCL2 expression. Results There was no significant correlation between BCL2 protein expression and overall survival within the GCB subgroup, but BCL2 expression had a significant adverse effect on overall survival within the ABC subgroup (P = .008). This correlation was also observed at the mRNA level (P < .04). The difference remained significant when the analyses were performed at different cutoff values. The t(14;18) was frequently observed in the GCB subgroup and was highly associated with BCL2 expression. Patients with ABC DLBCL did not exhibit t(14;18) but had a markedly higher frequency of chromosome 18q21 amplification, on which BCL2 resides. Thus, alternative mechanisms such as 18q21 amplification or activation of the nuclear factor-kappa B pathway, as reported previously, seem to be mainly responsible for the upregulation of BCL2 expression in the ABC subgroup. Conclusion Treating all DLBCL as a single entity ignores the mechanistic differences in BCL2 upregulation and obscures the prognostic significance of BCL2 expression. Hence, the significance of BCL2 and other biomarkers should be assessed in the context of DLBCL subgroups in future studies.

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LMO2 and Bcl-6 Expression in Diffuse Large B-Cell Lymphoma Predict Overall Survival in R-CHOP Treated Patients

Blood ◽

10.1182/blood.v112.11.5304.5304 ◽

2008 ◽

Vol 112 (11) ◽

pp. 5304-5304

Author(s):

Heather D. Brooks ◽

Robert M. Graham ◽

Randall L. Woodford ◽

Audrey K. Bennett ◽

Xin Qun Wang ◽

...

Keyword(s):

Overall Survival ◽

B Cell ◽

Germinal Center ◽

Cell Lymphoma ◽

Prognostic Significance ◽

Previous History ◽

B Cell Lymphoma ◽

Large B Cell Lymphoma ◽

Significant Difference ◽

Large B Cell

Abstract LMO2, a germinal center marker and transcription factor with an important role in angiogenesis and erythropoiesis, was found to confer improved prognosis in diffuse large B-cell lymphoma (DLBCL) when expressed in tissue microarray (TMA) samples (Natkunam et al J Clin Oncol2008,26:447). Bcl-6, a transcriptional repressor controlling germinal center formation, has been associated with favorable outcomes in DLBCL patients treated with CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) but was found to have no prognostic significance in DLBCL patients older than 60 treated with rituximab (R) + CHOP (Winter et al Blood2006,107:4207). Hans previously described an algorithm to subclassify DLBCL into germinal center B-cell like (GCB) and activated B-cell like (ABC) using immunohistochemical staining for CD10, bcl-6 and MUM1 with GCB exhibiting improved survival (Hans et al Blood2004,103:275). The aim of this study was to evaluate the prognostic value of LMO2, CD10, bcl-6, bcl-2 and MUM1 expression in DLBCL patients treated with R-CHOP. We also applied the Hans classification to our cohort and incorporated LMO2 into the algorithm. Adults (>18 years) with DLBCL who were diagnosed and treated at the University of Virginia between 2000 and 2007 were retrospectively evaluated. Patients were selected based on adequate tissue, treatment with at least 3 cycles of R-CHOP and if a minimum of 6 months of follow-up data was available at the time of data analysis. Forty-one cases were included of which 26 were de novo DLBCL, 12 were DLBCL with concurrent follicular lymphoma (FL) and 3 cases were new diagnoses of DLBCL with a previous history of FL. H&E-stained sections from paraffin-embedded, formalin-fixed blocks were used to define diagnostic areas, and 3 representative 1.0-mm cores were obtained from each case. TMA sections were then stained with antibodies to CD10, bcl-6, bcl-2, MUM1 and LMO2. Cases were assigned to GCB-like and ABC-like subgroups based on the Hans algorithm. The LMO2 stain was incorporated into the Hans algorithm to create modified-Hans (m-Hans) subgroups; the sample received an m-GCB designation if LMO2 positive but the normal Hans classification was applied if the sample was LMO2 negative. For 41 complete patient samples, a nonparametric Kaplan-Meier estimator was used to estimate overall survival (OS) probability for each group (m-GCB, m-ABC). A log-rank test demonstrated that there was a significant difference in OS probability between m-ABC and m-GCB groups (p = 0.047). Six month and 3 year survival for m-ABC and m-GCB were 71% and 88%, and 44% and 68% respectively. A semi-parametric Cox proportional hazard model was used to assess association between OS and each individual stain (CD10, bcl-2, bcl-6, MUM1 and LMO2), and a hazards ratio was used to quantify the effect of each stain. Bcl-6 and LMO2 expression were highly correlated, with bcl-6 expression significantly associated with OS (p=0.011). Patients without bcl-6 expression had more than four-fold risk than those expressing bcl-6 in OS (Hazard Ratio = 4.29, 95% CI: 1.4–13.1). Also, LMO2 expression showed a marginal association with OS (p=0.065). In conclusion, LMO2 incorporated into the Hans classification and bcl-6 expression are both significant predictors of OS in DLBCL patients treated with R-CHOP. Figure Figure

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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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Prediction of Survival in Diffuse Large B-Cell Lymphoma Based On the Expression of Two Genes: Integration of Tumor and Microenvironment Contributions.

Blood ◽

10.1182/blood.v114.22.622.622 ◽

2009 ◽

Vol 114 (22) ◽

pp. 622-622 ◽

Cited By ~ 1

Author(s):

Ash A Alizadeh ◽

Andrew J Gentles ◽

Alvaro J. Alencar ◽

Holbrook E Kohrt ◽

Roch Houot ◽

...

Keyword(s):

Gene Expression ◽

Overall Survival ◽

B Cell ◽

Tumor Cells ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Cell Of Origin ◽

Bivariate Model ◽

Large B Cell Lymphoma ◽

Large B Cell

Abstract Abstract 622 Background: Several gene-expression signatures are predictive of prognosis in diffuse large-B-cell lymphoma (DLBCL), but the lack of practical methods for a genome-scale analysis has restricted their routine clinical applicability. Methods: We studied genes whose expression had been reported to predict survival in DLBCL, attempting to validate genes and prognostic models with robust survival associations that are amenable to rapid diagnostic testing. Results: Among a previously described set of 6-genes shown to predict survival independent of measurement platform or therapy era (Lossos, et al. 2004 NEJM 350:1828), we identified LMO2 as the single gene with strongest independent prognostic value in 3 independent cohorts of patients with DLBCL. To assess the independent contribution of other genes in predicting survival, using existing microarray gene expression data (Lenz, et al. 2008 NEJM 359:2313), we evaluated all pairwise models that included LMO2 toward construction of a robust bivariate survival predictor. Among 54674 possible models, one combining expression of LMO2 with TNFRSF9 (encoding 4-1BB, also known as CD137) emerged as among the best in cross-validation when assessed in training (n=233) and test (n=181) cohorts. This bivariate predictor remained prognostic in both CHOP (p=1.7e-6) and R-CHOP (p=6.5e-8) therapy eras, was highly independent of the International Prognostic Index, Cell-of-Origin classification, 6-gene predictive model, ‘stromal' model, and added significantly to their prognostic power. While LMO2 expression was highly restricted to tumor cells and was linked to Cell-of-Origin (GCB, p=2.2e-16), TNFRSF9 expression was highest in non-tumor cells (P=0.02), particularly in an activated subset of infiltrating CD8 T-cells. To validate this bivariate model and devise a practical diagnostic assay, we used quantitative real-time polymerase-chain-reaction to measure the expression of LMO2 and TNFRSF9 as well as other components of the 6-gene model (BCL2, BCL6, FN1, CCL3, and CCND2) in diagnostic formalin fixed and paraffin-embedded samples of lymphoma from an independent set of 147 patients with de novo DLBCL treated with R-CHOP. The IPI distribution for these patients was: 0-1 factor (n=70), 2 factors (n=40), 3 factors (n=26), ≥4 factors (n=11). In univariate and multivariate analyses of this independent cohort, LMO2 and TNFRSF9 expression remained individually prognostic of both progression free and overall survival. The bivariate model combining LMO2/TNFRSF9 could be used to stratify distinct risk groups for overall survival (p=0.004), and remained independent of IPI. Conclusion: Measurement of the expression of two genes integrating contributions of tumor cells and the tumor microenvironment is sufficient to predict overall survival in patients with DLBCL treated with R-CHOP. Disclosures: Advani: Seattle Genetics, Inc.: Research Funding.

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Prognostic Significance of BACH2 Expression in Diffuse Large B-Cell Lymphoma: A Study of the Osaka Lymphoma Study Group

Journal of Clinical Oncology ◽

10.1200/jco.2005.02.1626 ◽

2005 ◽

Vol 23 (31) ◽

pp. 8012-8017 ◽

Cited By ~ 27

Author(s):

Emiko Sakane-Ishikawa ◽

Shin-ichi Nakatsuka ◽

Yasuhiko Tomita ◽

Shigeki Fujita ◽

Itsuko Nakamichi ◽

...

Keyword(s):

Overall Survival ◽

B Cell ◽

Cell Lymphoma ◽

Prognostic Significance ◽

B Cell Lymphoma ◽

Large B Cell Lymphoma ◽

Level 1 ◽

Level 2 ◽

Disease Free ◽

Large B Cell

Purpose BACH2, a B-cell–specific transcription repressor, is abundantly expressed in lymphocytes of B-cell lineage as well as B-cell lymphoma cell lines. BACH2 possesses an inhibitory effect on proliferation of Raji cell lines derived from Burkitt's lymphoma. In this study, the prognostic significance of BACH2 expression was examined in diffuse large B-cell lymphoma (DLBCL). Patients and Methods BACH2 expression was immunohistochemically examined on the paraffin-embedded sections obtained by biopsy from 108 patients (62 males and 46 females; age range, 23 to 85 years) with DLBCL. Staining intensity in the cytoplasm of the tumor cells was categorized as equal to or stronger (level 1) or weaker (level 2) than that in the endothelial cells in the same specimens. Results Level 1 and 2 expression of BACH2 was found in 32.4% and 67.6% of patients, respectively. Patients with level 1 expression showed significantly better disease-free and overall survival rate than those with level 2 expression (both P < .05). Multivariate analysis revealed BACH2 expression level together with performance status, elevated serum level of lactate dehydrogenase, and treatment response to be independent factors for prognosis of the patients. Conclusion BACH2 expression level is a useful marker to predict disease-free and overall survival of patients with DLBCL.

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