Survival Of Diffuse Large B-cell Lymphoma Patients Research Articles

Characterizing Autoimmune Disease-Associated Diffuse Large B Cell Lymphoma in a SEER-Medicare Cohort

Introduction: Severe immune dysregulation as seen in human immunodeficiency virus infection, immunosuppression after solid organ transplant, and autoimmune (AI) disease is known to act as a major risk factor for non-Hodgkin lymphoma (NHL). Recently, the InterLymph Subtypes Project pooled cases and controls to provide well-powered comparisons of risk factors for specific NHL subtypes, such as diffuse large B cell lymphoma (DLBCL), and showed that B cell-activating AI diseases in general (odds ratio 2.4; 95% confidence interval 1.8-3.1) and systemic lupus erythematosus (SLE) and Sjögren’s syndrome (SS) in particular were strongly associated with increased DLBCL risk after controlling for all other risk factors (Cerhan J Natl Cancer Inst Monogr. 2014). However, little is known about the demographics or clinical outcomes of DLBCL that arises in the setting of AI disease. We examined the Surveillance, Epidemiology, and End Results (SEER)-Medicare linked database to determine the frequency of common B cell AI diseases among older patients with DLBCL and characterize the patterns of presentation, treatment, and survival for DLBCL patients with concomitant AI disease.Methods: We used the SEER database for patients diagnosed 2002-2009 linked to their Medicare claims data through 2011 to characterize presentation, treatment, and survival patterns in patients with DLBCL, including those with rheumatoid arthritis (RA), SLE, SS, and other B cell-mediated autoimmune diseases as defined by InterLymph criteria (autoimmune hemolytic anemia, Hashimoto's thyroiditis/hypothyroidism, myasthenia gravis; pernicious anemia; Wang Am J Epidemiol. 2015). Patient age, sex, race/ethnicity, region of residence, marital status, year of diagnosis, cause of death, census tract-level characteristics (education, poverty, and metropolitan/urban/rural status), stage, B symptoms, and nodal or extranodal primary site of involvement were identified from the SEER data. Assessments of poor performance status, anemia, the Charlson comorbidity index, survival, and treatment strategies were identified using Medicare inpatient, outpatient and physician claims. Treatments were categorized as: rituximab (R), cyclophosphamide and vincristine (CVP), R-CVP; cyclophosphamide, hydroxydaunorubicin, and vincristine (CHOP), and R-CHOP. Patients who did not receive any DLBCL-directed treatment within 6 months of diagnosis were categorized separately. We examined the baseline clinical characteristics for patients with DLBCL and RA, SLE, SS, or any B cell-mediated autoimmune disease, plotted overall survival and lymphoma-related survival for these groups and compared median survival times.Results: Patient characteristics are summarized in the Table. With the exception that patients with DLBCL and AI disease were more commonly female[KJL1] [CF2] , patients with DLBCL and RA, SLE, SS, or other B cell AI diseases have similar baseline presenting features as other DLBCL patients and received similar first line treatments. There was a trend towards decreased lymphoma-related survival in patients with SLE and DLBCL compared to all other groups, but this difference was not statistically significant in this cohort (Figure) [KJL3] [KJL4] .Conclusions: In this retrospective claims-based cohort of older patients with DLBCL, concomitant AI disease was uncommon and was more likely to occur in female DLBCL patients, which likely reflects the higher incidence of AI disease in women. The possibility of lower lymphoma-related survival for these patients should be explored in future studies. [Display omitted] [Display omitted] DisclosuresFlowers:Millenium/Takeda: Research Funding; NIH: Research Funding; TG Therapeutics: Research Funding; Pharmacyclics, LLC, an AbbVie Company: Research Funding; Mayo Clinic: Research Funding; Infinity: Research Funding; ECOG: Research Funding; Acerta: Research Funding; AbbVie: Research Funding; Roche: Consultancy, Research Funding; Genentech: Consultancy, Research Funding; Gilead: Consultancy, Research Funding.

A population-based study of cellular markers in R-CHOP treated diffuse large B-cell lymphoma patients

Aim: To determine the prognostic significance of co-expression of MYC, BCL-2 and BCL-6 proteins in combination with other biomarkers and clinical characteristics within a population-based cohort of diffuse large B-cell lymphoma (DLBCL) patients uniformly treated with R-CHOP.Patients and methods: The immunohistochemical (IHC) expression of CD10, BCL-2, BCL-6, MUM1, MYC, CD5, CD30, Ki-67 and p53 was evaluated in a retrospective, population-based study comprising 188 DLBCL patients treated with R-CHOP and diagnosed in Sweden between 2002 and 2012.Results: Patients had a median age at diagnosis of 64 years (26–85 years) with a male:female ratio of 1.4:1. Approximately half (52%) of the patients presented with an International Prognostic Index (IPI) age adjusted (IPIaa) ≥ 2. Median follow-up time was 51 months (range 0.4–158) and the five-year lymphoma-specific survival (LSS) was 76%, five-year overall survival (OS) was 65% and five-year progression-free survival (PFS) was 61%. A high Ki-67 value was found in 59% of patients, while p53 overexpression was detected in 12% of patients and MYC, BCL-2 and BCL-6 expression were detected in 42%, 55% and 74% of patients, respectively. IPIaa ≥2 (p = 0.002), Ki-67 ≥ 70% (p = 0.04) and p53 overexpression ≥50% (p = 0.02) were associated with inferior LSS and OS. Co-expression of both MYC (>40%) and BCL-2 (>70%) proteins was detected in 27% of patients and correlated with a significantly inferior LSS (p = 0.0002), OS (p = 0.009) and PFS (p = 0.03). In addition, triple expression of MYC, BCL-2 and BCL-6, also correlated with a significantly inferior LSS (p = 0.02).Conclusion: Concurrent expression of MYC and BCL-2 proteins, as detected by IHC, was strongly associated with an inferior survival in DLBCL patients treated with R-CHOP. Other markers affecting survival were triple expression of MYC, BCL-2 and BCL-6, IPIaa, high Ki-67 and p53 overexpression.

Antiviral therapy improves overall survival in hepatitis C virus-infected patients who develop diffuse large B-cell lymphoma.

Chronic Hepatitis C virus (HCV) infection is associated with increased incidence of non-Hodgkin lymphoma. Several studies have demonstrated regression of indolent lymphoma with antiviral therapy (AVT) alone. However, the role of AVT in HCV-infected patients with diffuse large B-cell lymphoma (DLBCL) is unclear. We therefore analyzed AVT's impact on oncologic outcomes of HCV-infected patients (cases) who developed DLBCL. Cases seen at our institution (June 2004-May 2014) were matched with uninfected counterparts (controls) and then divided according to prior AVT consisting of interferon-based regimens. We studied 304 patients (76 cases and 228 controls). More cases than controls had extranodal (79% vs. 72%; p = 0.07) and upper gastrointestinal (GI; 42% vs. 24%; p = 0.004) involvement. Cases never given AVT had DLBCL more refractory to first-line chemotherapy than that in the controls (33% vs. 17%; p = 0.05) and exhibited a trend toward more progressive lymphoma at last examination compared to controls (50% vs. 32%; p = 0.09) or cases given AVT (50% vs. 27%; p = 0.06). Cases never given AVT had worse 5-year overall survival (OS) rates than did the controls (HR, 2.3 [95% CI, 1.01-5.3]; p = 0.04). Furthermore, AVT improved 5-year OS rates among cases in both univariate (median [Interquartile range]: 39 [26-56] vs. 16 [6-41] months, p = 0.02) and multivariate analyses (HR = 0.21 [95% CI, 0.06-0.69]; p = 0.01). This study highlights the negative impact of chronic HCV on survival of DLBCL patients and shows that treatment of HCV infection is associated with a better cancer response to chemotherapy and improves 5-year OS.

Abstract 3187: Concordance of immunohistochemistry (IHC) assay results with gene expression profiling (GEP) methods for diffuse large-B-cell lymphoma (DLBCL) subtype identification

Abstract Introduction: The survival of DLBCL patients who receive immunochemotherapy is correlated with the molecular signature of their tumors. The two most common subtypes which are defined by cell of origin (COO) are the germinal center B cell-like (GCB) and activated B cell-like (ABC) subtypes which have 3-year OS rates of 80% and 45%, respectively when treated with R-CHOP. Several methods have been developed to categorize DLBCL into these sub-types as the treatment they receive directly affects their outcome. Microarray GEP was first used to classify DLBCL into ABC or GCB, with a minority unclassified (UNC), however this is difficult to implement widely and has been replaced by more recent technologies. The most commonly used IHC-based Hans method profiles patients into GCB and non-GCB. The primary focus of this study was to evaluate concordance between current GEP and IHC methods to designate DLBCL subtypes. Methods: For this study, the IHC assay used was a standardized IHC assay based on the Hans algorithm and for the GEP-based assay; the NanoString Technologies Lymphoma Subtyping Test (LST) was used. Commercially available DLBCL samples (N = 138) were confirmed for tumor content and sent to a central laboratory for evaluation by the IHC assay. RNA was extracted from each sample and sent to NanoString for GEP evaluations. To compare methods homogeneously, calls for each method were converted to either “GCB” or “non-GCB”. All “ABC” and “UNC” subtype calls from the NanoString platform was converted to “non-GCB” and any “did not pass” (DNP) calls were converted to not available (NA). All NA values were excluded from the tables and concordance calculations. Results: The Positive Percent Agreement [95% CI] in calling non-GCB between IHC and LST was 87% [78% - 96%] (47/54). Overall concordance between IHC and LST was 80% [73% - 87%] (N = 125, 12 LST samples DNP, 1 IHC sample DNP). The Negative Percent Agreement between IHC and LST (i.e., in calling the GCB sub-type) was 75% [65% - 85%] using the LST result as a non-reference standard (53/71). Conclusions: Given that the success of targeted therapy for the treatment of DLBCL is contingent on the correct identification of sub-type of DLBCL, it is important to adopt a method that accurately and consistently identifies the sub-type in the majority of patients. Given the good concordance between the GEP and IHC methods, DLBCL can be stratified into COO sub-types using either platform. The results show that an optimized Hans IHC assay standardized in a central laboratory offers a robust, relatively inexpensive and accessible platform for cell of origin sub-typing in DLBCL and, with clinical validation, may offer an excellent tool for the selection of optimal therapy. Citation Format: Michael Schaffer, Shalini Chaturvedi, Sandy Frans, Regina Aquino, Jaehong Park, Brett Hall, Sriram Balasubramanian. Concordance of immunohistochemistry (IHC) assay results with gene expression profiling (GEP) methods for diffuse large-B-cell lymphoma (DLBCL) subtype identification. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3187.

The impact of FcγRIIa and FcγRIIIa gene polymorphisms on responses to RCHOP chemotherapy in diffuse large B-cell lymphoma patients.

Rituximab is a monoclonal antibody routinely used in the treatment of B-cell non-Hodgkin lymphomas. It mediates antibody-dependent cellular cytotoxicity of B lymphocytes by bridging them with Fcγ receptors (FcγR) on effector cells. Several polymorphisms in the FcγR genes have been identified to influence rituximab binding to FcγR, thus altering its antitumor effect in indolent lymphomas. In the present study, the impact of FcγRIIa and FcγRIIIa polymorphisms on the survival and response to immunochemotherapy consisting of rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone was evaluated in diffuse large B-cell lymphoma (DLBCL) patients. A total of 29 Slovenian DLBCL patients were studied. Genotyping was conducted for FcγRIIa-27, FcγRIIa-131, FcγRIIIa-48 and FcγRIIIa-158 polymorphisms. The median follow-up time was 29.7 months (range, 9.7–45.4 months). No significant impact of the genotypes was observed on the treatment response, progression-free or overall survival of DLBCL patients. There was a non-significant trend of an improved response to chemotherapy without additional irradiation in patients homozygous for Val at FCγIIIa-158 compared to Phe carriers. The findings of the present study indicate that FcγR polymorphisms have no influence on the survival of DLBCL patients.

Glasgow Prognostic Score Is Superior to Other Inflammation-Based Scores in Predicting Survival of Diffuse Large B-Cell Lymphoma

Inflammation-based prognostic scores, such as the glasgow prognostic score (GPS), prognostic index(PI), prognostic nutritional index(PNI), neutrophil lymphocyte ratio(NLR), platelet lymphocyte ratio(PLR) was related to survival in many solid tumors. Recent study showed that GPS can be used to predict outcome in diffuse large B-cell lymphoma(DLBCL). However other inflammation related scores had not been reported in DLBCL, and it also remained unknown which of them was more useful to evaluate the survival in DLBCL. In this retrospective study, a number of 252 newly diagnosed and histologically proven DLBCL patients from January 2003 to December 2014 were included. An elevated GPS, PI, NLR, PNI and PLR were all associated with decreased overall survival(P =0.000, P =0.000, P =0.006, P =0.001 and P =0.001, respectively) and event-free survival (P =0.000, P =0.000, P =0.011, P =0.001 and P =0.009, respectively) in univariate analysis. Multivariate analysis indicated that GPS(RR=1.768, 95%CI=1.043-3.000, p =0.034) remained an independent prognostic predictor in DLBCL. The area under the curve of GPS (0.735, 95%CI=0.645-0.824) was greater than that of PI(0.710, 95%CI=0.621-0.799), PNI(0.600, 95%CI=0.517-0.683), NLR(0.572, 95%CI=0.503-0.642), and PLR(0.599, 95%CI=0.510-0.689) by Harrell's C-statistics. Especially in the DLBCL patients treated with R-CHOP, GPS also remained the most powerful inflammation-based prognostic score when comparing with PI, NLR, PNI and PLR (P =0.004, P =0.000, respectively for OS and EFS). In conclusion, these results indicate that Inflammation-based prognostic scores such as GPS, PI, NLR, PNI and PLR can be used to evaluate the outcome in DLBCL patients. Among them, GPS is the most powerful tool in predicting survival in DLBCL patients, even in the rituximab era. DisclosuresNo relevant conflicts of interest to declare.

Systematic Analysis of Cell-of-Origin, Sequencing and Genomic Imbalances Identifies a Distinct Subset of Rituximab Treated Diffuse Large B-Cell Lymphoma with an Inferior Survival

Introduction: Diffuse Large B-cell Lymphoma (DLBCL), exhibits a wide range of clinical and biological heterogeneity. While initially responsive to rituximab based therapy, more than 70% of patients relapse within 5 years. Two distinct subtypes of DLBCLs were previously identified by gene expression profiling, that correlated to the cell of origin (COO); currently an immunohistochemistry (IHC) surrogate (Hans algorithm) is used to identify this COO. However, the value of COO identification by IHC is being questioned in patients treated with rituximab based regimen. As such there is a need for the identification of additional prognostication markers. The purpose of the present study is to systematically characterize a cohort of patients with DLBCL using a variety of methods including COO by IHC, copy number changes by array-based comparative genomic hybridization, and single gene mutations by next generation sequencing. The goal is to identify prognostic molecular biomarkers that predict survival better than current methods such as COO in DLBCL patients treated with rituximab.Methods: Patients diagnosed between 2003 and 2011 and treated with R-CHOP/R-EPOCH at BIDMC were identified. IPI/R-IPI, ECOG performance status, overall survival data were collected from retrospective chart review. Molecular and immunohistochemical studies were performed on formalin-fixed paraffin embedded tissue (FFPE), which was obtained prior to initiation of therapy (at the time of diagnosis).aCGH: Using a previously developed assay for aCGH to detect genomic gain/loss from archival FFPE, we characterized each DLBCL sample for the presence or absence of 50 copy number variations (CNVs) from 32 common regions of overlapping genomic imbalances comprising 36 minimal common regions. The calling criteria were based on GISTIC defined peaks based on copy number data from three publicly available datasets: IS-172, IS-51HR, EMEXP-3463.Gene panels: Next Generation Sequencing (NGS) was performed using a targeted hybrid capture panel and run on a Miseq (Illumina, Inc.). Gene selection for the panel was based on frequently mutated genes reported in DLBCL, Follicular lymphoma (FL), and Mantle Cell Lymphoma (MCL). Also selected for the panel were genes involved in known dysregulated pathways, therapeutic targets, and genes mapped to sites of genomic gains or losses.Cell of Origin: Immunostains were performed for BCL6, CD10, MUM1, FOXP1, GCET and LMO2. The immunostains were blindly scored by three different pathologists. For this study, GCB vs ABC determination was made using the Hans algorithm (CD10, BCL6 and MUM1 expression).Statistics: Kaplan-Meier (KM) survival analysis was performed using R (version 3.2.1).Results: Data on an initial subset of 49 patients with comprehensive molecular, COO and clinical information is presented (and an additional ~100 case analysis is in progress). Average age of patients at the time of diagnosis was 63, with 23 males and 26 females. By univariate analysis, the COO was not significantly associated with survival in the patients treated with rituximab based regimen (KM p=0.678). By contrast, three of the CNVs were associated with survival by Kaplan Meier analysis (Table 1). Also four, gene mutations were significantly associated with poor survival by univariate analysis (Table 1). Table 1Markers significantly associated with survival in rituximab treated patients with DLBCL. Ab28 predicted for better survival, whereas the presence of the other mutations or CNVs predicted for poor survival.Gene / CRp value (KM)Ab29 (D1p13.1)0.00787PIK3C2G0.00126PIM10.00499CD79B0.00567DTX10.0102Ab3(A1q21.1-q25.1(3))0.0193Ab28 (D1p36.32-p36.31)0.0211Conclusions: Our systematic analysis of DLBCLs using multiple immunohistochemical and molecular methods identified mutational and copy number biomarkers predictive of survival in DLBCL patients, treated with rituximab in univariate analysis. Multivariate analysis and data from additional cases will reveal whether these molecular biomarkers can better predict patient survival, compared to current methods (such as COO by IHC). DisclosuresFriedman:Cancer Genetics Inc.,: Employment, Equity Ownership. Guttapalli:Cancer Genetics, Inc.: Employment, Equity Ownership. Thodima:Cancer Genetics, Inc.: Employment, Equity Ownership. Kamalakaran:Cancer Genetics, Inc.: Employment, Equity Ownership. Houldsworth:Cancer Genetics Inc.,: Employment, Equity Ownership.

A PER3 polymorphism is associated with better overall survival in diffuse large B-cell lymphoma in Mexican population.

Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of malignant lymphoma. Presently, one of the most important clinical predictors of survival in DLBCL patients is the International Prognostic Index (IPI). Circadian rhythms are the approximate 24 hour biological rhythms with more than 10 genes making up the molecular clock. Determine if functional single nucleotide polymorphism in circadian genes may contribute to survival status in patients diagnosed with diffuse large B-cell lymphoma. Sixteen high-risk non-synonymous polymorphisms in circadian genes (CLOCK, CRY2, CSNK1E, CSNK2A1, NPAS2, PER1, PER2, PER3, PPP2CA, and TIM) were genotyped by screening PCR. Results were visualized by agarose gel electrophoresis and confirmed by two-direction sequencing. Clinical variables were compared between mutated and non-mutated groups. LogRank survival analysis and Kaplan-Meier method were used to calculate the overall survival. PER3 rs10462020 variant showed significant difference in overall survival between patients containing mutated genotypes and those with non-mutated genotypes (p = 0.047). LDH levels (p = 0.021) and IPI score (p < 0.001) also showed differences in overall survival. No clinical differences were observed in mutated vs. non-mutated patients. This work suggests a role of PER3 rs10462020 in predicting a prognosis in DLBCL overall survival of patients.

Upregulation of long noncoding RNA PEG10 associates with poor prognosis in diffuse large B cell lymphoma with facilitating tumorigenicity.

Diffuse large B cell lymphoma (DLBCL) is one of the most common malignancies worldwide. To date, there has been little progress in improving the overall survival of DLBCL patients. Emerging evidences have implicated that long noncoding RNAs (lncRNAs) have important regulatory roles in fundamental biological processes, and some of them are involved in cancer initiation, development and progression. This study was to investigate the expression of lncRNA PEG10 in a cohort of DLBCL patients to assess its clinical value and biological function in DLBCL. We first found that the expression of PEG10 was upregulated in DLBCL tumorous tissues and that cell lines compared with the normal. Moreover, we illustrated that PEG10 was significantly correlated with B symptoms, IPI score, CHOP-like treatment and rituximab. In addition, ROC(AUC) of PEG10 was up to 0.8228, implicating that PEG10 could be a diagnostic marker for distinguishing DLBCL from normal. Importantly, we verified that PEG10 was a key independent predictive factor for DLBCL prognosis from sizable samples through the longtime follow-ups. Furthermore, we revealed that knockdown of PEG10 expression by siRNA could lead to growth arrest and cell apoptosis in vitro. Our results suggested that PEG10 could represent a novel indicator of poor prognosis and might be served as a potential target for the diagnosis and gene therapy of DLBCL.

The Prognostic Role and Relationship between E2F1 and SV40 in Diffuse Large B-Cell Lymphoma of Egyptian Patients.

Diffuse large B-cell lymphoma (DLBCL) is the most common type of lymphomas worldwide. The pathogenesis of lymphomas is not yet well understood. SV40 induces malignant transformation by the large T-antigen (L-TAG) and promotes transformation by binding and inactivating p53 and pRb. L-TAG can bind pRb promoting the activation of the E2F1 transcription factor, thus inducing the expression of genes required for the entry to the S phase and leading to cell transformation. This immunohistochemical study was conducted to assess the prognostic role and relationship of SV40 L-TAG and E2F1 in diffuse large B-cell lymphoma (DLBCL) of Egyptian patients. This retrospective study was conducted on 105 tissue specimens including 20 follicular hyperplasia and 85 DLBCL cases. SV40 L-TAG was identified in 3/85 (4%) of DLBCL. High Ki-67 labeling index (Ki-67 LI) and apoptotic count were associated with high E2F1 expression (p<0.001 for all). No significant association was reached between E2F1 and SV40. E2F1 expression proved to be the most and first independent prognostic factor on overall survival of DLBCL patients (HR = 5.79, 95% CI = 2.3–14.6, and p<0.001). Upregulation of E2F1 has been implicated in oncogenesis, prognosis, and prediction of therapeutic response but is not seemingly to have a relationship with the accused SV40.

CXCR4 expression enhances diffuse large B cell lymphoma dissemination and decreases patient survival.

The chemokine receptor CXCR4 has been implicated in the migration and trafficking of malignant B cells in several haematological malignancies. Over-expression of CXCR4 has been identified in haematological tumours, but data concerning the role of this receptor in diffuse large B cell lymphoma (DLBCL) are lacking. CXCR4 is a marker of poor prognosis in various neoplasms, correlating with metastatic disease and decreased survival of patients. We studied CXCR4 involvement in cell migration in vitro and dissemination in vivo. We also evaluated the prognostic significance of CXCR4 in 94 biopsies of DLBCL patients. We observed that the level of expression of CXCR4 in DLBCL cell lines correlated positively with in vitro migration. Expression of the receptor was also associated with increased engraftment and dissemination, and decreased survival time in NOD/SCID mice. Furthermore, administration of a specific CXCR4 antagonist, AMD3100, decreased dissemination of DLBCL cells in a xenograft mouse model. In addition, we found that CXCR4 expression is an independent prognostic factor for shorter overall survival and progression-free survival in DLBCL patients. These results show that CXCR4 mediates dissemination of DLBCL cells and define for the first time its value as an independent prognostic marker in DLBCL patients.

Hepatitis C Infection Is Associated with Hepatic Toxicity but Does Not Compromise the Survival of Patients with Diffuse Large B Cell Lymphoma Treated with Rituximab-Based Chemotherapy

Introduction: The influence of hepatitis C virus (HCV) infection on outcome of patients with diffuse large B cell lymphoma (DLBCL) treated with rituximab-based chemotherapy is still controversial.Materials and Methods: We retrospectively analyzed the characteristic and clinical outcomes of patients with DLBCL newly diagnosed between Jan 2005 and Dec 2011. All patients were treated with at least one course of rituximab-containing chemotherapy. The clinical characteristics between groups (29 HCV-positive or 139 HCV-negative) were compared using a Chi-square method for categorical variables, a Student’s t-test for continuous variables, and a non-parametric method (Wilcoxon Sum-Rank test) for AST, AST and total bilirubin. Univariate analysis followed by multivariate analysis was used to identify the variables associated with hepatotoxicity.Results: The medium follow-up duration was 3.0 (0.07-8.02) years. HCV infection resulted in more hepatic toxicity in both univariate (55.2% vs. 23.0%, P = 0.001) and multivariate (P = 0.003) analyses. In addition, HCV-positive DLBCL patients were more likely to have treatment delay (20.1% vs. 0.7%, P = 0.004). For patients who developed hepatic toxicity during immunochemotherapy, HCV-positive patients had significantly higher folds of aspartate aminotransferase elevation (3.39 folds vs. 2.06 folds, P = 0.042) and total bilirubin elevation (1.60 folds vs. 0.96 folds, P = 0.012) compared with those who were HCV negative. However, HCV did not influence the 5-year progression-free survival rate (P = 0.412) or 5-year overall survival rate (P = 0.410).Conclusions: HCV infection is associated with increased incidence and severity of hepatic toxicity and delayed chemotherapy without compromised survival in DLBCL patients treated with rituximab-based chemotherapy. DisclosuresNo relevant conflicts of interest to declare.

High Body Mass Index (BMI) in North American Elderly Diffuse Large B-Cell Lymphoma (DLBCL) Patients Treated with Rituximab (R)-CHOP Compensates for Negative Impact of Male Gender

Background: Recent reports from prospective clinical trials of R-containing chemotherapy in DLBCL patients suggest that gender, weight and/or BMI influence clinical outcomes. Pharmacokinetic studies by the German High Grade Lymphoma Study Group have shown that R clearance is relatively slow in elderly women compared to men, leading to higher levels and prolonged exposure and hence better clinical outcomes in elderly females. Specifically, it has been suggested that elderly men are underdosed, based on faster R clearance (Muller et al., 2012; Pfreundschuh et al., 2014). Regarding BMI as a predictor of clinical outcome, analysis of the US Veterans Administrative database showed an association between increased BMI and improved survival in DLBCL patients (Carson et al., 2012), while the ECOG clinical trial (E4494) for elderly DLBCL patients failed to reveal a significant association of BMI with clinical outcomes, or a gender difference related to BMI in failure-free survival (Hong et al., 2014). To further investigate these associations, we studied the effect of gender, BMI as well as body surface area (BSA, the actual dosing parameter), and potential interactions among these factors on long-term clinical outcomes for elderly DLBCL patients in the National Comprehensive Cancer Network (NCCN) non-Hodgkin lymphoma database.Methods:De novo DLBCL patients with age > 60 yrs. were identified from the NCCN adult DLBCL cohort. Patients were diagnosed between June 2000 and December 2010. All received R as part of first-line therapy. Outcomes evaluated included progression free survival (PFS) and overall survival (OS) at 3 years based on patient gender, age and BMI/ BSA at presentation. Gender was stratified based on BMI (<=18.5, >18.5-25, >25) or BSA (<=2, >2), and Kaplan-Meier estimates were calculated. Associations with disease progression and survival were additionally adjusted for the International Prognostic Index (IPI) in the multivariable Cox regression analyses.Results: Of the 1,386 DLBCL patients who received R, 627 were elderly with age >60 yrs. The majority of elderly men were either overweight or large: only 13% had BMI <=25 and only 26% had BSA <=2. Elderly men (n=325, 52%) experienced worse PFS (3 yr-Hazard Ratio, HR 1.5, 95% CI: 1.1-2.1, p=0.02) and OS (3 yr-HR 1.6, 1.1-2.4, p=0.01) compared to elderly women. Of note, the poor risk associated with male gender was associated with patients over 60, but not the younger cohort (3 yr-PFS, HR 1.3, 0.9-1.9, p=0.12). The benefit associated with female gender in the elderly cohort decreased with increases in BMI and BSA (Figure). There was a benefit associated with female gender when compared to male gender in the BMI (<=25) (log rank, p<0.01) and BSA <=2 (p=0.04) strata, but not among patients with high BMI (>25) or BSA (>2). In multivariable analysis, low or normal BMI as compared to high BMI was independently associated with poor outcomes (3-yr PFS, HR: 1.6, 1.1-2.2, p<0.01) after adjusting for gender. There was a trend suggesting that BSA <=2 correlated with worse 3-yr PFS in the elderly group adjusting for gender (3 yr-HR 1.4, 0.9-2.0, p=0.12). The HR estimates remained largely unchanged after adjusting for IPI. Notably, higher BMI was not associated with more favorable prognostic clinical factors.Conclusions: Our results, derived from analysis of unselected patients with DLBCL treated with R-containing chemotherapy at major NCCN centers, confirmed an age-dependent disadvantage to male gender in treatment outcomes. The magnitude of this negative effect diminished with higher levels of BMI and BSA, with the greatest negative impact occurring in elderly men with BMI <=25 or BSA <=2. Our findings support efforts to optimize R dosing, especially in the elderly male subset with BMI <=25 or BSA <=2. Future prospective trials should factor size and gender into the study design and analysis. Our results support the ongoing German randomized trial to evaluate enhanced rituximab dosing for older male patients. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

The Stimulatory Role of CD300a in Diffuse Large B-Cell Lymphoma

CD300a is a type I transmembrane receptor protein which has shown inhibitory effect on B-cell receptor mediated signals. In an analysis of publicly available data on diffuse large B-cell lymphoma (DLBCL), however, we found that the expression levels of CD300A mRNA were inversely correlated with the overall survival of DLBCL patients. When analyzing the transcript levels of CD300a in human tissues, we found that CD300a mRNA levels were significantly greater in DLBCL tissues than benign lymphoid tissues (P<0.05). To decipher the role of CD300a in DLBCL, we used shRNA system to knock-down the expression levels of CD300a in DLBCL cell lines, and found that decreased levels of CD300a significantly inhibited cell proliferation of OCI-Ly1 cells, but not of VAL, OCI-Ly10 or SUDHL-8 cells. Mechanistically, reduced expression of CD300a resulted in a marked attenuation of Akt phosphorylation in OCI-Ly1 cells. Pharmacologic inhibition of PI3K by LY294002 displayed a similar inhibitory effect on cell proliferation, indicating the possible involvement of PI3K/Akt signaling pathway in CD300a’s effect. Furthermore, using a xenograft animal model, we found that decreasing expression levels of CD300a in OCI-Ly1 cells significantly inhibited tumor formation of these cells in vivo. Collectively, our results suggested a stimulatory role of CD300a in DLBCL which could serve as a potential biomarker and therapeutic target for this malignance. DisclosuresNo relevant conflicts of interest to declare.

Hepatitis C infection is associated with hepatic toxicity but does not compromise the survival of patients with diffuse large B cell lymphoma treated with rituximab-based chemotherapy

The influence of hepatitis C virus (HCV) infection on the outcome of patients with diffuse large B cell lymphoma (DLBCL) treated with rituximab-based chemotherapy is controversial. We retrospectively analyzed the characteristics and clinical outcomes of 168 patients with DLBCL diagnosed between January 2005 and December 2011. Twenty-nine patients who were HCV-positive before lymphoma treatment were compared with 139 patients who did not have HCV infection. The median follow-up duration was 3.0 (0.07–8.02) years. HCV infection resulted in more hepatic toxicity in both univariate (p=0.001) and multivariate (p=0.003) analyses. In addition, HCV-positive DLBCL patients were more likely to have treatment delay (20.1% vs. 0.7%, p<0.001). For patients who developed hepatic toxicity during immunochemotherapy, HCV-positive patients had significantly higher folds of aspartate aminotransferase elevation (p=0.042) and total bilirubin elevation (p=0.012) compared with those who were HCV negative. However, HCV did not influence the 5-year progression-free survival rate (p=0.412) or 5-year overall survival rate (p=0.410). In conclusion, HCV infection is associated with increased hepatic toxicity and delayed chemotherapy without compromised survival in DLBCL patients treated with rituximab-based chemotherapy.

FOXP1 directly represses transcription of proapoptotic genes and cooperates with NF-κB to promote survival of human B cells

AbstractThe forkhead transcription factor FOXP1 is involved in B-cell development and function and is generally regarded as an oncogene in activated B-cell–like subtype of diffuse large B-cell lymphoma (DLBCL) and mucosa-associated lymphoid tissue lymphoma, lymphomas relying on constitutive nuclear factor κB (NF-κB) activity for survival. However, the mechanism underlying its putative oncogenic activity has not been established. By gene expression microarray, upon overexpression or silencing of FOXP1 in primary human B cells and DLBCL cell lines, combined with chromatin immunoprecipitation followed by next-generation sequencing, we established that FOXP1 directly represses a set of 7 proapoptotic genes. Low expression of these genes, encoding the BH3-only proteins BIK and Harakiri, the p53-regulatory proteins TP63, RASSF6, and TP53INP1, and AIM2 and EAF2, is associated with poor survival in DLBCL patients. In line with these findings, we demonstrated that FOXP1 promotes the expansion of primary mature human B cells by inhibiting caspase-dependent apoptosis, without affecting B-cell proliferation. Furthermore, FOXP1 is dependent upon, and cooperates with, NF-κB signaling to promote B-cell expansion and survival. Taken together, our data indicate that, through direct repression of proapoptotic genes, (aberrant) expression of FOXP1 complements (constitutive) NF-κB activity to promote B-cell survival and can thereby contribute to B-cell homeostasis and lymphomagenesis.

Clinicopathological implications of nuclear factor κB signal pathway activation in diffuse large B-cell lymphoma.

Although abnormal activation of the nuclear factor κB (NF-κB) signaling pathway plays an important role in the pathogenesis of diffuse large B-cell lymphoma (DLBCL), only a few studies have dealt with the relation of NF-κB activation to clinical outcomes in this disease. We analyzed the clinical characteristics of 147 consecutive DLBCL patients, examined paraffin-embedded tissues from 120 of them to identify the activation of the NF-κB pathway by using immunohistochemical staining, and performed an overall survival (OS) analysis. Expression of P-p65 and p52 was found in 30.0% (n = 36) and 35.8% (n = 43) of the patients, respectively. Coexpression of these factors was found in 16.7% (n = 14) of the cases. Patients were divided into 4 groups according to P-p65 and/or p52 expression: P-p65(+) only, p52(+) only, both P-p65(+) and p52(+), and both P-p65(-) and p52(-). The 3-year OS rates in the 4 groups were 51.3%, 68.3%, 34.6%, and 85.8%, respectively (P = .006). Univariate analysis showed that early stage (P = .032), low International Prognostic Index score (P = .001), less than 2 extranodal metastases (P = .014), complete remission with chemotherapy (P < .0001), germinal-center B-cell-like subtype (P = .049), Ki-67 < 75% (P = .017), and P-p65(-) (P = .002) or p52(-) (P = .031) were associated with longer 3-year OS. Multivariate analysis indicated that P-p65 expression was an independent prognostic factor for shorter OS (P = .032). In conclusion, NF-κB pathway activation markers P-p65 and p52 predict poor survival in DLBCL patients.

Deregulated expression of miR-224 and its target gene: CD59 predicts outcome of diffuse large B-cell lymphoma patients treated with R-CHOP.

miRNAs are non-coding RNA molecules; their deregulations may contribute to cancer pathogenesis. However, the mechanisms of how miRNA dysfunction contributes to the lymphomagenesis of diffuse large B-cell lymphoma (DLBCL) are not well established. In this study, we analyzed the expression of miR-224 in four DLBCL cell lines and 168 patients' specimens. We found that the expression of miR-224 in DLBCL was down-regulated compared with normal B-cell but was not statistically different between the germinal center B-cell-like-type and the activated B-cell-like-type. Using bioinformatics prediction and luciferase report assays, we demonstrated that miR-224 directly down-regulated CD59 expression by binding to its 3'-untranslated region. We also used immunohistochemical staining of CD59 in human DLBCL specimens and analyzed the relationship between the expression of miR-224, CD59 and the overall/progress-free survival of DLBCL patients who were uniformly treated with rituximab,cyclophosphamide, adriamycin, vincristine, and prednisone (R-CHOP). We found that miR-224 may contribute to DLBCL pathogenesis. Most importantly, the expression of miR-224 and CD59 can predict the response and outcome of DLBCL patients treated with R-CHOP.

Diffuse large B-cell lymphoma with histone H3 trimethylation at lysine 27: another poor prognostic phenotype independent of c-Myc/Bcl2 coexpression.

Deregulation of histone H3 trimethylation at lysine 27 (H3K27me3) via aberration of the histone methyltransferase, enhancer of zeste homologue 2 (EZH2), is suggested to play a critical role in cancers including hematologic malignancies. In the present study, implications of H3K27me3 were investigated in diffuse large B-cell lymphoma (DLBCL) with respect to clinicopathological factors, especially in association with c-Myc/Bcl2 coexpression and germinal center B-like (GCB) or non-GCB subtype. By immunohistochemistry, a high level of H3K27me3 was observed in approximately one-third (35.3%, 79/224) of DLBCL cases, and this subset of cases was related to poor performance status (Eastern Cooperative Oncology Group scores ≥ 2) (P = .013), elevated lactate dehydrogenase level (P = .001), and a higher international prognostic index risk group (scores ≥3) (P = .005). H3K27me3 level was significantly correlated with EZH2 expression (P = .004) and c-Myc protein expression (P = .003) but not correlated with c-Myc/Bcl2 coexpression or with GCB or non-GCB subtype. A high level of H3K27me3 was related to an inferior overall survival (P = .006) and was shown to be an independent prognostic factor for overall survival along with the higher international prognostic index risk group and c-Myc/Bcl2 coexpression. In conclusion, H3K27me3 was related to EZH2 and c-Myc expression, suggesting formation of a MYC-EZH2-H3K27me3 loop in a subgroup of DLBCL cases. H3K27me3 was associated with poor patient outcome and revealed as an independent predictor for overall survival of DLBCL patients. H3K27me3 in DLBCL may be another high-risk phenotype independent of the phenotype of c-Myc/Bcl2 coexpression or other known poor prognostic subgroups.

The rate of reduction in the maximum standardized uptake value from the initial to the post-R-CHOP therapy in positron emission tomography scan predicts disease progression in diffuse large B cell lymphoma patients

[18F]fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) is useful for primary staging and evaluation of treatment outcome in diffuse large B cell lymphoma (DLBCL) patients. The reduction in the maximum standardized uptake value (ΔSUVmax) from the initial to the interim 18F-FDG PET scan has been reported to predict survival in DLBCL patients. We retrospectively evaluated ΔSUVmax obtained by PET or PET-computed tomography before and after initial therapy in 31 newly diagnosed DLBCL patients who were treated with R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone) therapy. Receiver observation characteristic curve analysis was used to evaluate the optimal cutoff value for the ΔSUVmax for disease progression. The 3-year progression-free survival rate of patients with ΔSUVmax≥83 and <83% was found to be 91 and 25%, respectively (P<0.001). The 4-year overall survival rate of patients with ΔSUVmax≥83 and <83% was found to be 100 and 83%, respectively (P=0.046). The ΔSUVmax observed before and after R-CHOP therapy could be useful in the prediction of disease progression and survival in newly diagnosed DLBCL patients.