Outcome In Diffuse Large B-cell Lymphoma Research Articles

Clinicopathologic features and outcomes of diffuse large B-cell lymphoma with extranodal involvement: A retrospective analysis

Background: Extranodal involvement is seen in about 40%–50% of the cases of diffuse large B-cell lymphoma (DLBCL). However, data on the clinicopathological features and outcomes of DLBCL with extranodal involvement, especially in the rituximab era, are scarce from India.Objectives: In this study, we aimed to assess the clinicopathological features and outcomes of DLBCL with extranodal involvement among Indian patients.Materials and Methods: This retrospective study was conducted on patients with DLBCL, registered in the Department of Medical Oncology of the All India Institute of Medical Sciences, Delhi, India, between January 2014 and December 2018. Patients with nodal DLBCL aged more than 18 years, with concomitant extranodal disease in any anatomical site were included. All patients were treated with multiagent chemotherapy with or without rituximab.Results: Median age of the cohort was 50 years (range, 18–86), a male-to-female ratio of 2:1. B-symptoms and bulky disease were seen in 48% and 39% patients, respectively. Bone was the most common site of extranodal involvement observed in 32% patients. Bone marrow involvement was present in 13% patients. Low-risk International Prognostic Index (IPI) was seen in 32% patients and intermediate risk IPI in 38% patients. Cyclophosphamide, doxorubicin, vincristine, and prednisolone (CHOP)-based treatment was used in 80% of the patients and rituximab (R) was used in 73% of the patients. The objective response rate was 76%, with a complete response rate of 65.5%. After a median follow-up of 26 months, the 3-year progression free survival and overall survival (OS) were 65% and 82.7%, respectively. Involvement of the kidney/adrenal gland and central nervous system, high IPI score, and use of non- R-CHOP regimens were associated with a poor OS on the multivariate analysis.Conclusions: The bone is the most common site of extranodal involvement in Indian patients with DLBCL. The outcome of extranodal DLBCL depends on the primary site of disease involvement.

Current Frontline Treatment of Diffuse Large B-Cell Lymphoma.

Diffuse large B-cell lymphoma (DLBCL), the most common subtype of non-Hodgkin lymphoma, is an aggressive and biologically heterogeneous disease. Risk stratification and treatment algorithms vary based on stage of disease and bulk along with other clinical and biological factors, including the International Prognostic Index, cell of origin, and other molecular subsets. Rituximab (Rituxan), cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) is the current standard of care and cures more than 60% of patients. The role of radiotherapy is largely restricted to patients with limited-stage disease. In elderly patients, geriatric assessments of baseline fitness and functional status help optimize therapy based on the balance of efficacy and toxicity. While numerous randomized trials have failed to improve upon R-CHOP, a recent press release from the POLARIX trial (NCT03274492) suggests that adding polatuzumab vedotin (Polivy) to rituximab, cyclophosphamide, doxorubicin, and prednisone (pola-R-CHP) improves progression-free survival and may replace R-CHOP in eligible patients. Ongoing trials are exploring frontline therapy that integrates other novel agents, including various small molecules, bispecific antibodies, and chimeric antigen receptor T-cell therapy, with promising preliminary results. Defining a population of patients with high-risk disease in whom R-CHOP is not effective is critical. Patient selection based on refining molecular subsets, quantitative PET metrics such as metabolic tumor volume, and dynamic risk assessments using interim PET and circulating tumor DNA analysis may allow for a personalized, response-adapted approach that will further improve outcomes in DLBCL.

No difference in treatment outcome between patients with nodal versus extranodal diffuse large B-cell lymphoma

Diffuse large B-cell lymphoma (DLBCL) has been classified using various parameters, including the site of origin. Studies have reported conflicting outcomes when DLBLC patients were stratified according to the site of origin. This study aimed to investigate the response rate and survival outcomes in nodal versus extranodal DLBCL and compare the results to a region-matched study covering the 1988 - 2005 period. A single-center retrospective cohort study was conducted on all patients diagnosed with DLBCL and treated in a tertiary care hospital in Pakistan during 2014 - 2019. We calculated the mean and median for continuous variables and frequency and percentages for all categorical variables. Progression-free survival (PFS) and overall survival (OS) were calculated using Kaplan-Meier survival curves. A Cox proportional hazards model was used to determine the hazard ratio (HR) for OS. Of the 118 patients, 49 patients (41.5%) had nodal disease and 69 patients (58.5%) were diagnosed with extranodal DLBCL. The majority of patients in the nodal and extranodal cohorts presented with Stages III and IV disease (73.4% and 62.3%, respectively). A complete response to (immuno) chemotherapy was achieved in 71.4% of nodal DLBCL patients and 65.2% of extranodal DLBCL patients. The 5-year PFS and median PFS in the entire cohort were 0.8% and 17 m, respectively. The PFS and median PFS in the nodal and extranodal DLBCL cohort were 0% and 1.4%, respectively, and 15 m and 19 m, respectively. The 5-year OS and median OS in the entire cohort were 16.1% and 19 m, respectively. The OS and median OS in the nodal and extranodal DLBCL cohort were 8.2% and 21.7%, respectively, and 19 m and 21 m, respectively. Multivariable linear regression revealed that the ABC phenotype (nodal, HR = 1.37, 95% CI = 1.37 - 3.20; extranodal, HR = 1.65, 95% CI = 1.46 - 3.17; GBC as reference) and double and triple hit DLBCL (nodal, HR = 1.29, 95% CI = 1.19 - 2.81; extranodal, HR = 1.87, 95% CI = 1.28 - 2.43; and non-expressors as reference) are independent negative predictors of OS. DLBCL incidence in the Karachi region has remained comparable but patient composition in the extranodal DLBCL cohort has shifted to predominantly advanced stage. Nodal and extranodal DLBCL were associated with similar PFS and OS profiles and first- and second-line treatment responses. Cell of origin and antigen expression status was independent negative predictors of OS, disfavoring the ABC phenotype and lesions with c-MYC and BCL2 and/or BCL6 overexpression. DLBCL is an aggressive type of non-Hodgkin's lymphoma, however; patients respond well to standard systemic chemotherapy. Extranodal type of DLBCL patients tend to have more residual disease after first-line systemic chemotherapy, but physicians should keep in mind that the subsequent line treatment mitigates its negative impact on survival.

Diffuse large B-Cell lymphoma: from novel molecular classifications to tailored targeted therapies

Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease comprising multiple genetic subtypes that translates and impacts clinical outcomes after standard chemoimmunotherapy. Our initial understanding of the complex biological subtypes of DLBCL began with the identification of cell of origin (COO), and now has evolved to include even more specific subtypes defined by genetic signatures and mutations. These newer classifications lend themselves to the application of precision-based medicine, allowing us to tailor new treatment platforms that target specific oncogenic drivers in order to improve DLBCL outcomes. Essential to this is the development of genetic assays and tools that are reliable and readily available to assist in the application of these molecular classifications to real-world use. In this review, we discuss the history of DLBCL classification systems and their implication on clinical investigation as well as novel therapeutic options in DLBCL.

Mir-671-5p, Mir-193b-5p, Mir-1307-5p Are Useful for Predicting Outcome in Diffuse Large B-Cell Lymphoma

Background: Diffuse large B-cell Lymphoma (DLBCL) is stratified histologically as GCB and ABC phenotypes based on Hans algorithm. Besides this categorization, there is a paucity of molecular markers useful for predicting response to CHOP/R-CHOP chemotherapy and outcome of DLBCL patients. We performed miRNA profiling to identify potential miRNAs for their ability to predict response to chemotherapy and outcome in DLBCL patients.Methods: A total of 41 cases of histologically and immunohistochemically confirmed DLBCL cases who received R-CHOP/CHOP were included in this study after IRB approval. The follow up ranged from 7 to 2273 days with a mean of, 874 days. Cases in sustained complete remission for ≥ 1095 days were identified as “responders” whereas cases with partial response/stable disease or primary progressive disease were identified as“non-responders”. In the first part of the study, miRNA transcriptome profiling was performed from lymph node tissue of 10 cases of DLBCL which included 3 responders and 7 non-responders (3 with refractory disease and 4 with relapse) for identification and selection of differentially expressed miRNAs. Total RNA was isolated from the biopsies, libraries constructed with ION TOTAL RNA-SEQ v2 and sequenced on S5 ION Torrent. The filtered reads were aligned to the reference genome hg19 using Bowtie. Differentially expressed miRNAs were identified by DESeq2. In the second part of the study, validation of the selected miRNAs was performed in a retrospective validation cohort of 41 cases including 17 responders and 24 non-responders by semi-quantitative qRT-PCR. ROC curves were obtained and best cut-off for the delta Ct values was selected for each of the miRNAs to distinguish the two cohorts of patients. In the third part of the study, an additional 38 cases of DLBCL were included prospectively as testing cohort, in whom mi-RNA were isolated from paired pre-treatment biopsy and plasma samples were evaluated for the expression of the selected miRNAs for prediction of response to treatment and outcome.Results: miRNA transcriptome profiling revealed a total of 30 differentially expressed miRNAs among DLBCL responders vs non-responders and the data was submitted to GEO database (GSE179760). The top 4 significantly downregulated miRNAs in non-responders selected for validation included miR-193b-5p, miR-671-5p, miR-1307-5p and miR-326 as shown in the volcano plot and heatmap in Fig.1. In-silico prediction for the interacting genes of the selected miRNAs revealed genes involved in lymphoma progression and chemoresistance . Validation by qRT-PCR revealed significant down-regulation of miR-1307-5p, miR-671-5p and miR-193b-5p in the non-responders compared to responders (Fig.2). There was no significant association of miRNA expression with disease stage, nodal or extranodal origin and the presence of B symptoms. However, the downregulation of miR-671-5p was found to be associated with the presence of B-symptoms. Downregulation of miR-193b-5p, miR-671-5p, miR-1307-5p expression showed a significant association with progression free survival as shown in Fig.3 (Spearman's correlation plot and Kaplan-Meier survival curves).In the prospective cohort of 38 DLBCL patients, all 3 miRNAs were expressed in cell free plasma and tissue but only miRNA-193b-5p showed correlation between them. Using the cut-off Ct values obtained from ROC curves of tissue miRNA levels in the validation cohort , 16/38 patients were predicted to be non-responders. The preliminary data showed that 8 of these 16 cases were correctly predicted (6 died of disease, 2 showed partial response). The remaining cases are still under follow-up. The treatment response could not be predicted on the corresponding plasma levels.Conclusion: Downregulation of miRNA-671-5p, miR-193b-5p and miR-1307-5p in pre-treatment biopsy samples is associated with poor outcome and shorter progression free survival and poor treatment response leading to refractory/relapse DLBCL, which needs confirmation in larger studies. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Obesity Is Associated with Increased Relapse Risk in Diffuse Large B-Cell Lymphoma

Introduction: Diffuse Large B-Cell Lymphoma (DLBCL) is an aggressive Non-Hodgkin's lymphoma (NHL) characterized by clinical, biological and genetic heterogeneity. Despite the tremendous advancement in anticancer therapy against DLBCL, up to 40% of patients are refractory and/or relapse after first-line chemo-immunotherapy. Mechanisms responsible for chemo-immunotherapy failure are largely uncharacterized; however, it is believed the microenvironment contributes to tumor adaptation and survival. Several studies have shown that high body mass index (BMI) is associated with increased risk of developing DLBCL and obesity is an independent prognostic indicator of worse outcome when compared to non-obese patients. Weight gain alters the metabolic landscape of tumor microenvironment to inhibit T cell function and promote cellular growth. Obesity induced proinflammatory mediators can activate numerous signaling pathways including nuclear factor (NF)-KB that result in anti-apoptotic and increased proliferative efficiency in B-cells Here, we report the results of our retrospective study from two hospital systems in Houston, Texas evaluating the impact of BMI on progression-free-survival (PFS) in DLBCL patients receiving standard induction chemotherapy.Methods: After Institutional Review Boards (IRB) approval, patients (pts) were identified by electronic medical record database query. Inclusion criteria included: (a) age>18 years old, (b) DLBCL diagnosis confirmed by pathology review at Ben Taub Medical Center and Baylor St. Luke's Medical Center. BMI was calculated using patient's height and weight at diagnosis. For all comparison, we selected DLBCL pts with World Health Organization (WHO) predefined overweight, class I, II, III obesity to better capture effect of extreme BMI on DLBCL PFS. Cell-of-origin (COO) was determined by Hans classification. R-CHOP/R-EPOCH were classified as intense therapy whereas R-CVP/R-CEOP as non-intense therapy. Descriptive statistics was used to evaluate differential expression of clinical and laboratory variables. PFS among pts with BMI <35 and BMI >35 was estimated with Kaplan Meier Method. Cox regression model evaluated effect of independent variables on PFS for pts with DLBCL exhibiting obesity. SAS was used for all analysis.Results: 123 pts were identified. Demographic, clinical and laboratory data was balanced among DLBCL pts with and without BMI >35 (Table 1). Median BMI was 25.9 for pts with BMI<35 v 37.3 in pts with BMI >35. In the pts with BMI <35, 36.9% were classified as germinal center b-cell (GCB) subtype and 63.1% were classified as non-GCB subtype (p= NS). Within the BMI >35, 46.2% were GCB subtype and 53.9% were non-GCB subtype. 45.9% of pts had low International Prognostic Index (IPI) score in BMI<35 compared to 56.52% in BMI>35, whereas 54% had high IPI in BMI<35 compared to 43.48% in BMI>35. PFS was 2,864 days vs 763 days in patients with BMI <35 and >35, respectively (p=0.02). (Fig 1). To address effect of “type of therapy administered” (low v high intensity), COO, and age as confounders for our observation that PFS is inferior in DLBCL pts with BMI >35, Cox regression model was performed confirming the independent predictive role for BMI >35 only on DLBCL PFS.Conclusions: Our study demonstrates that obesity (BMI >35) results in increased relapse risk in DLBCL patients. In our population, risk of relapse was enhanced by higher BMI independent of age, therapy administered and COO. Obesity, by itself, especially BMI>35 (Class II and Class III) may represent a unique factor for adverse outcome in DLBCL. The mechanism for which extreme obesity leads to inferior PFS in DLBCL is unclear. It is possible that obesity induces “high risk” mutations acquisition and/or adversely modifies tumor microenvironment resulting in chemotherapy cytotoxic attenuation. Larger studies are needed to externally validate our observation. [Display omitted] DisclosuresMims: Biogen: Current holder of individual stocks in a privately-held company; Incyte: Research Funding; IDEC: Current holder of individual stocks in a privately-held company; Celgene: Research Funding; Pfizer: Research Funding; AVEO: Research Funding.

The Impact of Marginalization on Diffuse Large B-Cell Lymphoma Overall Survival: A Retrospective Cohort Study

Introduction: While health disparities in diffuse large B-cell lymphoma (DLBCL) have been previously noted, literature systematically describing the impact of social determinants of health (SDOH) on DLBCL overall survival (OS) is sparse. Furthermore, existing data largely examine SDOH of health in isolation, not accounting for key covariates or disease-related variables. Marginalization, which accounts for various SDOH, is a process of systemic discrimination and exclusion. Given the paucity of literature to date, we examined how marginalization influences DLBCL OS in the Canadian setting. The objectives of this study were: 1) To describe the impact of marginalization on DLBCL OS and 2) To identify which dimensions of marginalization, if any, impacted OS.Methods: We conducted a population-based retrospective cohort study of adult patients with newly diagnosed DLBCL in Ontario between January 1, 2005 to December 31, 2017 receiving a rituximab-containing chemotherapy regimen for curative intent, followed until March 1, 2020. Our primary exposure of interest was the Ontario Marginalization Index (ON-Marg), an administrative database tool which combines demographic indicators into four distinct marginalization dimensions: residential instability (% renters and % living alone), material deprivation (% low income and % lone parent families), dependency (% seniors and % employment), and ethnic concentration (% recent immigrants and % visible minority). Our primary outcome was 2-year OS, defined as time from the date of first rituximab-based treatment to date of death or completion of follow-up. Survival curves were generated using Kaplan-Meier methods, and Cox regression analyses were used to identify covariates that were independently associated with OS. Our final model adjusted for age (as increments of 10 years), sex, and comorbidity burden as measured by aggregate diagnostic groups (ADGs).Results: A total of 10,344 patients were diagnosed with DLBCL and treated with a rituximab-containing regimen in Ontario between January 1, 2005 and December 31, 2017. The median age was 67 (IQR, 55-75) and 46% were female. Of patients who had staging data (49%), 54% were advanced stage at diagnosis, and the median number of cycles of chemoimmunotherapy received was 6 (4-6). Median number of ADGs was 10 (8-12), indicating a moderate-to-high burden of comorbidities within this cohort, and 13% of patients resided in a rural area. Cohort characteristics and mortality rates per ON-Marg quintile (Q5 = most marginalized) are provided in Table 1. Overall, group characteristics were evenly distributed, except that the most marginalized group had a higher proportion of patients residing in urban settings and nearer to treatment hospitals. Two-year overall survival was 73.2%. After controlling for relevant confounders, material deprivation (hazard ratio [HR] 1.06, 95% confidence interval [CI] 1.02 - 1.10, p=0.003) and ethnic concentration (HR 1.05, 95% CI 1.01 - 1.09, p=0.013) contributed to increased risk of all-cause mortality. Residential instability and dependency had no significant effect. Increasing age (HR 1.29, 95% CI 1.25 - 1.33, p<0.0001) and increasing number of ADGs (HR 1.028, 95% CI 1.025 - 1.031, p<0.0001) were also associated with increased risk of all-cause mortality, while female sex was found to be protective (HR 0.89, 95% CI 0.83 - 0.97, p=0.005).Discussion: To our knowledge, no study has examined the impact of marginalization and collective SDOH on DLBCL outcomes. We found that increased material deprivation and ethnic concentration conferred increased mortality. While Canada's largely single-payer system and previously documented healthy immigrant effect may have been expected to attenuate effects of income and immigration, our study suggests otherwise. Possible explanations may include barriers to healthcare access for patient subsets, who may have less support for treatment. Structural factors such as systemic racism, health literacy, and caregiver burden must also be taken into account. These factors require further evaluation to inform targeted interventions and establish support for increased access to timely care amongst this patient population. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Combining PET Radiomic Features with MYC Gene Rearrangement Results in High Prediction of Outcome in Diffuse Large B-Cell Lymphoma

IntroductionGenetic abnormalities, such as MYC oncogene rearrangements, contribute to the outcome heterogeneity in diffuse large B-cell lymphoma (DLBCL) patients. These rearrangements occur in 10-15% of DLBCL patients and have been associated with a poor prognosis. Recently, radiomics features extracted from PET/CT scans have shown to be predictive of outcome. The aim of this study was to investigate if the ability to predict outcome in DLBCL can be improved by combining different clinical, radiomics and genetic features.Methods323 DLBCL patients from the HOVON-84, HOVON-130, and PETAL trials with a baseline PET/CT scan and a minimum follow-up of two years were included. MYC status was assessed using Fluorescence in situ hybridization (FISH). 245 patients were MYC negative, whereas 25 patients had a MYC rearrangement and 57 patients had MYC and BCL2 and/or BCL6 rearrangements. Lesions were delineated using a semi-automated preselection of 18F-FDG avid structures defined by a SUV4.0 threshold using the ACCURATE tool. Next, 5 conventional PET features (maximum standardized uptake value (SUV max), SUV peak, SUV mean, metabolic tumor volume (MTV) and total lesion glycolysis and 18 dissemination features were extracted. Dissemination features were pertaining to distance between lesions, differences in uptake between lesions and differences in volume between lesions. Logistic regression with backward feature selection was used to predict 2-year time to progression, defined as time from baseline PET/CT to progression. We tested the predictive value of 4 models. 1) a clinical model using individual components of the international prognostic index (IPI): Ann Arbor stage (categorical), WHO performance status (categorical), lactate dehydrogenase (LDH) levels (dichotomous) and age (continuous), 2) a model that included clinical and genetic predictors: MYC status (categorical) and IPI components, 3) a model that included radiomics features: 5 conventional PET and 18 dissemination features and 4) a model that combined clinical and genetic predictors with radiomics features. Model performance was assessed using repeated cross-validation (5-fold, 1000 repeats) yielding the cross-validated area under the curve of the receiver-operator-characteristics curve (CV-AUC). To match prevalence of MYC-positive patients with real-world prevalence (Rosenwald et al, JCO 2019) all 245 MYC-negative patients were used for each repeat, and 10 MYC-FISH_positive DLBCL patients and 20 patients with MYC and BCL2 and/or BCL6 rearrangements were selected using random stratified sampling. Regression coefficients were averaged over all folds to calculate the probability of progression for all patients. High- and low-risk groups were defined based on prevalence of events and the diagnostic performance was assessed using positive- and negative predictive values.ResultsThe highest model performance for the clinical model was observed when combining Ann Arbor stage, LDH and extranodal involvement and yielded in a CV-AUC of 0.69 (95% confidence interval (CI): 0.52-0.83). MYC status combined with WHO performance status, LDH and extranodal involvement yielded an improved CV-AUC of 0.71 (95% CI: 0.52-0.86). The highest model performance for the radiomics model was observed for MTV combined with the maximum distance between the largest lesion and any other lesions (Dmax bulk), the maximum difference in SUV peak between two lesions (DSUVpeak patient) and the maximum difference in volume between two lesions (DVol patient) yielding a CV-AUC of 0.77 (95% CI: 0.62-0.90). The optimal combined model included MYC status, WHO performance status, LDH, MTV, Dmax patient, DSUVpeak patient and DVol patient after backward feature selection and yielded a CV-AUC of 0.77 (95% CI: 0.60 - 0.90). The positive predictive value was highest for the combined model (53.0%) and increased by 20% compared to the optimal clinical model (33.1%). Negative predictive values were comparable between models and ranged between 85.8-88.1%.ConclusionsPrediction models using 18F-FDG PET/CT radiomics features at baseline aids in identifying DLBCL patients at high risk for relapse. The positive predictive value increased when radiomics features were added to the clinical and genetic parameters. Therefore, radiomics features can increase the efficiency of clinical trials by only selecting poor prognosis patients. [Display omitted] DisclosuresChamuleau: Gilead: Research Funding; Genmab: Research Funding; Celgene: Research Funding. Lugtenburg: Incyte: Honoraria; Regeneron: Honoraria; Genmab: Honoraria; Servier: Honoraria, Research Funding; Takeda: Honoraria, Research Funding; Celgene: Honoraria, Other: Non-financial support; Travel support; Roche: Honoraria, Research Funding. Dührsen: CPT Cellex Patient Treatment: Honoraria, Membership on an entity's Board of Directors or advisory committees; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees. Hüttmann: Celgene: Honoraria; Gilead: Honoraria; Lead Discovery Center GmbH: Consultancy; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees. Richter: HTG Molecular Diagnostics, Inc.: Current Employment, Research Funding. Klapper: Regeneron: Consultancy, Research Funding; Amgen: Research Funding; Roche: Consultancy, Research Funding; Takeda: Consultancy, Research Funding. Zijlstra: Takeda: Research Funding.

The Dark Side of Pyroptosis of Diffuse Large B-Cell Lymphoma in B-Cell Non-Hodgkin Lymphoma: Mediating the Specific Inflammatory Microenvironment.

Background: Diffuse large B-cell lymphoma (DLBCL) is a common aggressive B-cell non-Hodgkin lymphoma (B-NHL). While combined chemotherapy has improved the outcomes of DLBCL, it remains a highly detrimental disease. Pyroptosis, an inflammatory programmed cell death, is considered to have both tumor-promoting and tumor-suppressing effects. The role of pyroptosis in DLBCL has been gradually appreciated, but its value needs further investigation. Methods: We analyzed mutations and copy number variation (CNV) alterations of pyroptosis-related genes (PRGs) from The Cancer Genome Atlas (TCGA) cohort and evaluated the differences in expression in normal B cells and DLBCL patients in two Gene Expression Omnibus (GEO) datasets (GSE12195 and GSE56315). Based on the expression of 52 PRGs, we divided 421 DLBCL patients from the GSE31312 dataset into distinct clusters using consensus clustering. The Kaplan-Meier method was used to prognosis among the three clusters, and GSVA was used to explore differences in the biological functions. ESTIMATE and single-sample gene-set enrichment analysis (ssGSEA) were used to analyze the tumor immune microenvironment (TME) in different clusters. A risk score signature was developed using a univariate survival analysis and multivariate regression analysis, and the reliability and validity of the signature were verified. By combining the signature with clinical factors, a nomogram was established to predict the prognosis of DLBCL patients. The alluvial diagram and correlation matrix were used to explore the relationship between pyroptosis risk score, clinical features and TME. Results: A large proportion of PRGs are dysregulated in DLBCL and associated with the prognosis. We found three distinct pyroptosis-related clusters (cluster A, B, and C) that differed significantly with regard to the prognosis, biological process, clinical characteristics, chemotherapeutic drug sensitivity, and TME. Furthermore, we developed a risk score signature that effectively differentiates high and low-risk patients. The nomogram combining this signature with several clinical indicators showed an excellent ability to predict the prognosis of DCBCL patients. Conclusions: This work demonstrates that pyroptosis plays an important role in the diversity and complexity of the TME in DLBCL. The risk signature of pyroptosis is a promising predictive tool. A correct and comprehensive assessment of the mode of action of pyroptosis in individuals will help guide more effective treatment.

Impact of germline polymorphisms in genes regulating glucose uptake on positron emission tomography findings and outcome in diffuse large B-cell lymphoma: results from the PETAL trial

Background[18F]Fluoro-deoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) is the standard imaging procedure in diffuse large B-cell lymphoma (DLBCL). Disease presentation, FDG-PET/CT performance, and outcome may be influenced by germline single nucleotide polymorphisms (SNP) in genes regulating glucose uptake.MethodsClinical variables, FDG-PET findings, and outcome were analysed in relation to SNPs in 342 DLBCL patients participating in the ‘Positron Emission Tomography-Guided Therapy of Aggressive Non-Hodgkin Lymphomas’ (PETAL) trial. Genes analysed included SLC2A1 (SNPs rs1385129, referred to as HaeIII; rs710218, HpyCH4V; rs841853, XbaI), VEGFA (rs3025039), HIF1A (rs11549465, P582S; rs11549467, A588T), and APEX1 (rs1130409, D148E). Statistical significance was assumed at p ≤ 0.05.ResultsThe SLC2A1 HaeIII and HpyCH4V SNPs were tightly linked and statistically significantly associated with baseline maximum standardized uptake value (SUVmax) and Ann Arbor stage, with slightly lower SUVmax (HaeIII, median 18.9, interquartile range [IQR] 11.5–26.6, versus 21.6, IQR 14.4–29.7; p = 0.019) and more frequent stage IV disease (HaeIII, 44.5% versus 30.8%; p = 0.011) in minor allele carriers. As previously reported for lung cancer, the association was dependent upon the coexistent APEX1 D148E genotype. The HIF1A A588T SNP was associated with total metabolic tumour volume (TMTV) and time-to-progression, with significantly lower TMTV (median 16 cm3, IQR 7–210, versus 146 cm3, IQR 34–510; p = 0.034) and longer time-to-progression in minor allele carriers (log-rank p = 0.094). Time-to-progression was also associated with the SLC2A1 XbaI and APEX1 D148E SNPs, with shorter time-to-progression in homozygous and heterozygous SLC2A1 XbaI (HR 1.456; CI 0.930–2.280; p = 0.099) and homozygous APEX1 D148E minor allele carriers (HR 1.6; CI 1.005–2.545; p = 0.046). In multivariable analyses including SNPs, International Prognostic Index factors, sex, and B symptoms, HIF1A A588T, SLC2A1 XbaI, and APEX1 D148E retained statistical significance for time-to-progression, and SLC2A1 XbaI was also significantly associated with overall survival.ConclusionsCommon SNPs in genes regulating glucose uptake may impact SUVmax, tumour distribution, tumour volume, and outcome in DLBCL. The effects on SUVmax are of low magnitude and appear clinically negligible. The results are consistent with findings in other types of cancer. They need to be confirmed in an independent DLBCL population of sufficient size.Trial registrationTrial registration: ClinicalTrials.gov NCT00554164; EudraCT 2006-001641-33. Registration date November 5, 2007, https://www.clinicaltrials.gov/ct2/show/NCT00554164

ABCL-300: The Relation Between Hepatitis C Infection and Cell of Origin (COO) of Diffuse Large B-Cell Lymphoma (DLBCL) and the Prognostic Value of Both Factors (Egyptian Experience)

Context COO has a significant prognostic impact on DLBCL outcome. HCV plays a great role in B-cell proliferation and transformation to NHL. Objective This study was conducted to emphasize the impact of COO classification on survival in DLBCL and its correlation to HCV infection. Design This prospective study was conducted on 50 cases of DLBCL. Setting This study was conducted on patients with DLBCL in OCMU in Egypt. Patients The mean age of our patients was 55 years, and 58% of them were female. Interventions DLBCL histopathological diagnosis was performed by evaluating hematoxylin and eosin-stained sections from formalin-fixed, paraffin-embedded tissue blocks from the tumor, combined with CD20 stain. COO was detected by using the Hans algorithm. RT-PCR for HCV infection was done for all patients. All patients received induction with the CHOP protocol. Main Outcomes Whether HCV-positive patients would have shorter survival. Results About 51.5% of non-GCB cases were stage IV in correlation to only 35.5% of GCB cases, with significant p-value (p-value = 0.002). Also, about 21% of non-GCB cases showed relapse in comparison to only 11.8% of GCB cases, with no significance. Therefore, non-GCB predicts a dismal outcome in NHL. In regard to the correlation between survival and COO, GCB cases showed significantly prolonged PFS and OS (78 and 88 months, respectively) in comparison to only 40.5 and 31 months for non-GCB cases (p-values = 0.054 and 0.024). There was a significant correlation between HCV positivity and attaining CR (p-value = 0.025). Also, HCV-positive patients showed shortened PFS and OS in comparison to HCV-negative cases, although not significant. In regard to the correlation between HCV status and COO, about 25% of GCB cases were HCV positive in comparison to 58.8% of non-GCB, with a p-value of 0.026. In multivariate analysis, GCB, low- and low-intermediate IPI, and CR achievement were considered independent predictors of survival (p-values = 0.004, 0.015, and 0.001, respectively). Conclusions The Hans algorithm is a good predictor of survival in B-NHL. HCV positivity has a poor impact on outcomes in NHL. There is possibly a correlation between HCV infection and COO for lymphoma, which needs further analysis.

Clinicopathological and prognostic features of hepatitis B virus-associated diffuse large B-cell lymphoma: a single-center retrospective study in China

BackgroundWhile the epidemiologic association between hepatitis B virus (HBV) infection and diffuse large B-cell lymphoma (DLBCL) is established, little is known about the pathological characteristics and outcome of DLBCL arising in patients with HBV infection.MethodsWe retrospectively studied a cohort of 420 patients with DLBCL for the incidence of HBV infection, and the clinicopathologic features and prognostic factors in HBsAg-positive DLBCL patients in China, a hepatitis B endemic area.ResultsIn our study, 127 (30.2%) patients were HBsAg-positive. HBsAg-positive DLBCL displayed a younger median onset age (50 vs. 54 years, P = 0.002), more frequent involvement of the spleen (19.7% vs. 6.1%, P < 0.001), less frequent involvement of the small and large intestine (2.3% vs. 11.2%, P = 0.003), more advanced disease (stage III/IV: 56.7% vs. 45.1%, P = 0.028), and lower expression rate of MYC (49.1% vs. 66.7%, P = 0.026). The median follow-up time was 61.9 months. Univariate analysis showed that there was no significant difference in overall survival (OS) between HBsAg-negative and -positive DLBCL (P = 0.577). In the HBsAg-positive DLBCL subgroup, age older than 60 years, advanced disease, elevated lactate dehydrogenase (LDH), spleen involvement, B symptoms (fever, night sweats, weight loss), and double expressers of MYC and BCL2 had a significantly worse outcome, and patients treated with R-CHOP (rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone) had a better prognosis. Multivariate analysis further confirmed that spleen involvement and rituximab use were independent prognostic factors in HBsAg-positive DLBCL patients.ConclusionsOur study indicates that HBsAg-positive DLBCL has unique clinicopathological features and independent prognostic factors. Moreover, under antiviral prophylaxis, the survival of DLBCL patients with HBV infections was comparable to that of HBV-negative patients, and the use of rituximab significantly improved OS in HBsAg-positive DLBCL patients.

Methylation silencing CDH23 is a poor prognostic marker in diffuse large B-cell lymphoma.

Cadherin-23(CDH23) mediates homotypic and heterotypic cell-cell adhesions in cancer cells. However, the epigenetic regulation, the biological functions, the mechanisms and the prognostic value of CDH23 in diffuse large B-cell lymphoma (DLBCL) are still unclear. The Gene Expression Profiling Interactive Analysis (GEPIA) and the Gene Expression Omnibus (GEO) database were employed to analyze the CDH23 expression level in DLBCL. The correlation of CDH23 expression and methylation was analyzed by LinkedOmics database. The prognostic value was analyzed via GEPIA. Correlated genes, target kinase, target miRNA, target transcription factor and biological functions were identified by LinkedOmics and GeneMANIA database. The relationship between CDH23 and the immune cell infiltration was explored by the Tumor Immune Estimation Resource (TIMER). The expression of CDH23 was reduced by DNA methylation significantly in DLBCL tissue. Reduction of CDH23 represented poor outcome of DLBCL patients. Functional enrichment analysis showed that CDH23 mainly enriched in cancer cell growth, cell metastasis, cell adhesion, cell cycle, drug catabolic process, leukocyte mediated immunity and DNA repair by some cancer related kinases, miRNAs and transcription factors. These results indicated that methylated reduction of CDH23 represented poor outcome of DLBCL. CDH23 is associated with essential biological functions and key molecules in DLBCL. CDH23 may play crucial roles in DLBCL tumorigenesis. Our results lay a foundation for further investigation of the role of CDH23 in DLBCL tumorigenesis.

Six Cycles of R-CHOP-21 Is Non-Inferior to 8 Cycles for Treatment of Diffuse Large B-Cell Lymphoma - a Nordic Lymphoma Group Population Based Study

Background:Standard treatment for patients with diffuse large B-cell lymphoma (DLBCL) consists of R-CHOP-21, established when two randomised trials did not find dose-dense administration of R-CHOP to be superior (Cunningham et.al 2013, Delarue et.al 2013). In both these trials, as well as in the majority of other clinical trials that provide the current evidence base for use of R-CHOP-21, 8 cycles of R-CHOP-21 were administered. An exception is the MInT-study, where 6 cycles of R-CHOP-21 was associated with superior outcome to 6 cycles of CHOP-21 for low-risk patients aged <60 (Pfreundschuh et.al 2006). Despite a lack of solid evidence to support the use of 6 instead of 8 cycles of R-CHOP-21 for all patients, administration of 6 cycles is commonly utilised in practice. Also, this strategy is supported by the European Society for Medical Oncology (ESMO) and National Comprehensive Cancer Network (NCCN) guidelines (Tilly et.al 2015, Zelenetz et.al 2016). The NCCN guidelines motivate the use of 6 cycles for all patients partly by the increased toxicity in absence of clinical benefit seen on administration of 8 cycles compared to 6 cycles of dose-dense R-CHOP-14 among patients aged 61-80 in the RICOVER-60 trial (Pfreundschuh et.al 2008).To our knowledge, no randomised trial has demonstrated superiority of 8 compared to 6 cycles of R-CHOP-21. As part of a current project on utilising real world data from the Danish and Swedish lymphoma registries to predict outcome in DLBCL, we examined whether patients treated with 6 or 8 cycles of R-CHOP-21 had similar survival and could be analysed as a single group.Methods: The study population was identified through the Swedish and Danish Lymphoma Registries 2007-2015. Patients with leptomeningeal-, eye- or CNS involvement were excluded. Data was conditioned to reach 200 days overall survival to minimise the risk of immortal time bias. Multivariate cox regression analysis included number of chemotherapy cycles administered, age, Ann Arbor stage, performance status, elevated lactate dehydrogenase (LDH), gender and involvement of more than one extranodal site. Sensitivity analyses with patients stratified according to low or high risk, age >/≤70 and excluding patients treated with consolidative radiotherapy were performed.Results:During the time-frame of this study, a total of 1200 adult patients received 6 or 8 cycles of R-CHOP-21 in Sweden and Denmark. Of these, 1037 (86%) were administered 6 cycles, and 163 (14%) received 8 cycles. Mean age was 70 years (range 18-91) in the cohort who received 6 cycles, and 68.5 years (range 25-90) for patients treated with 8 cycles. Radiotherapy was administered to 121 (10%) patients. A larger proportion of patients who received 8 cycles of R-CHOP-21 presented with Ann Arbor stage III-IV, performance status >1, elevated levels of LDH and thus a higher IPI-score (Table 1). Conditional 5-year overall survival rates were 73% in both treatment groups (Figure 1). No difference in outcome was detected for patients administered 6 versus 8 cycles of R-CHOP-21 in a multivariate analysis (hazard ratio: 0.86, 95% confidence interval: 0.62-1.20, p=0.36). Additionally, consistent results of no difference were observed in all patient subgroups stratified according to age (>/≤70), IPI (low/high) or no radiotherapy.Conclusion: Based on these real world data, we note that the majority of patients with DLBCL treated with R-CHOP-21 during the surveyed time period received 6 cycles, and conclude that the outcomes of patients treated with 6 cycles of R-CHOP-21 were non-inferior to the outcomes of patients treated with 8 cycles of R-CHOP-21.Figure 1. Overall survival for patients treated with 6 (red line) and 8 (green line) cycles. [Display omitted] DisclosuresJørgensen:Roche: Research Funding. Smedby:Janssen: Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees. El-Galaly:Takeda: Other: Travel funding; Roche: Other: Travel funding.

Gender-Based Differences in Efficacy of Immuno-Chemotherapy for Diffuse Large B-Cell Lymphoma Revisited: A Danish Population-Based Study

Background: The addition of CD20 antibodies (rituximab) to CHOP(-like) chemotherapy has been the single most important progress in the treatment of diffuse large B-cell lymphoma (DLBCL) for decades. Clinical studies suggested more pronounced outcome improvements following immuno-chemotherapy in female DLBCL as compared to male patients, which was supported by pharmacokinetic studies showing faster clearance of rituximab in elderly male patients.Aims: To explore gender-specific differences in DLBCL outcome in a population-based setting and to estimate the gender-specific improvements in relative survival seen with the introduction of immuno-chemotherapy for DLBCL.Patients and Methods: The study was based on the nationwide Danish lymphoma registry (LYFO), which contains detailed information on lymphoma patients treated at Danish hematology departments. Patients diagnosed in the period 2000-2014 and treated upfront with CHOP or CHOP-like therapy were analyzed according to the use of rituximab or not. During the surveyed time period, rituximab was given intravenously at a dose of 375mg/m2 synchronously with CHOP(-like) chemotherapy. A relative survival approach was applied using flexible parametric survival models (1) in which the expected mortality was given by age-, sex-, and year-matched lifetables from Statistics Denmark. The overall effect of rituximab was estimated by a model with two levels (rituximab, R+ vs. no rituximab, R-) and five degrees of freedom for the baseline excess hazard; based on this model 0-5 years relative survival and excess hazard ratio (EHR) were estimated. Gender-specific outcome improvements were estimated by expanding the model with gender as an effect modification of rituximab. The life-year gain attributable to rituximab in the first five years after diagnosis was estimated by the differences in loss in expectation of life (2) for male and female DLBCL patients treated with or without rituximab. All analyses were performed using Stata (StataCorp. 2015. Stata Statistical Software: Release 14 . College Station, TX: StataCorp LP).Results: In total, 4,368 DLBCL patients were included (3,321 in the R+ group and 1,047 in the R- group). Patient characteristics, treatment information, complete remission rates, and 5-year overall survival (OS) estimates are shown in Table 1. Men treated with rituximab were older than men without rituximab treatment suggesting a shift over time toward more intensive treatment for elderly. For both women and men the following was seen: higher Ann Arbor stage, higher R-IPI risk group, higher proportion of patients in CR, and longer 5-year OS when comparing patients with vs. without rituximab treatment. Figure 1A shows relative survival curves. Five-year relative survival of DLBCL patients was 67% (95% CI: 63% - 70%) for patients treated without rituximab and 77% (95% CI: 75% - 79%) for patients with rituximab, and the excess hazard ratio (EHR) was 1.47 (1.27 - 1.70) for patients treated without rituximab. Rituximab was also associated with substantial improvements in relative survival for male and female DLCBL patients analyzed separately (figure 1B-C). The gains of lifetime attributed to rituximab in the first 5 years after diagnosis were 0.26 years (95% CI: 0.10 - 0.43) and 0.40 years (95% CI: 0.21 - 0.61) for men and women, respectively. The effect modification of gender on rituximab was not statistically significant (p = 0.236).Conclusions: In the present study, there was no statistically significant effect modification of gender on rituximab with respect to relative survival. There was a trend toward marginally larger gain in lifetime for women after inclusion of rituximab when compared to men, although this difference was also not statistical significant and may be explained by other factors.

Mouse Lymphoma Model-Based Senescence Exploitation to Predict Treatment Outcome of DLBCL Patients

Introduction: Molecularly informed treatment decisions are the basis of personalized cancer precision medicine. While gene expression profiling has been used to map the molecular landscape of tumors and to study the impact of molecularly defined subtypes on treatment outcome, biological effector principles, specifically cellular senescence, remain largely understudied. Syngeneic mouse models of cancer that recapitulate core molecular features of human malignancies could serve as useful models to explore mechanisms of drug sensitivity, and, likewise, to dissect molecular mechanisms of treatment failure. In particular, we focus here on the role of therapy-induced senescence, a histone H3 lysine 9 trimethylation (H3K9me3)-governed terminal cell-cycle arrest program.Methods: The Eµ- myc transgenic lymphoma mouse was used as a cross-species model of chemoresistance for patients diagnosed with diffuse large B-cell lymphoma (DLBCL). Gene expression profiles (GEP) of primary Eµ- myc lymphomas were obtained at diagnosis and after exposure to cyclophosphamide chemotherapy in vivo, with long-term outcome monitored in a clinical trial-like fashion. Lymphoma senescence capacity, a pivotal drug effector principle, was interrogated in mice by unbiased approaches as well as genetic perturbations.Results: DLBCL-established gene expression-based classifiers related to cell-of-origin (COO - i.e. GCB/ABC) subtypes and distinct DLBCL biologies (e.g. comprehensive consensus clusters [CCC]) were instrumental to identify comparable lymphoma subsets in the murine system. In turn, mice carrying senescence-defective lymphomas due to engineered loss of the H3K9me3 methyltransferase Suv39h1 or overexpression of the H3K9-active demethylase LSD1 succumbed faster to chemotherapy, as DLBCL patients with high LSD1 or low H3K9me3 expression in their lymphomas presented with significantly inferior outcome. Likewise, cross-species application of senescence-associated characteristics identified in unengineered primary mouse lymphomas unveiled molecular features important for treatment outcome in human DLBCL. Ongoing mouse model-based work aims at specific targeting of aberrant H3K9 demethylase activity to assess therapeutic long-term effects and to determine the conditions for future early-phase clinical testing in DLBCL patients.Conclusions: Primary Eµ- myc transgenic lymphomas serve as a faithful model for human DLBCL, since they share key pathologic and genetic features, including CCC/COO-based clustering. In turn, mouse lymphoma-based molecular exploration, genetic dissection and functional interrogation of treatment effector mechanisms, which is difficult to conduct in patient material, can be re-applied to molecularly and clinically co-annotated datasets from DLBCL patients to inform novel, lesion- and state-driven personalized treatment approaches and the design of future clinical trials. DisclosuresNo relevant conflicts of interest to declare.

Low Circulating B Lymphocytes in Newly Diagnosed Follicular and Diffuse Large B Cell Lymphoma: Differences Compared to Normal Subjects and Possible Prognostic Role

Introduction: Reduction of the lymphocyte to monocyte ratio (LMR) has an unfavorable impact on overall survival in Diffuse Large B Cell Lymphoma (DLBCL), and on treatment response in Follicular Lymphoma (FL). Abnormal LMR seems primarily due to a reduction in absolute value of lymphocytes. Based on these observations, we have prospectively investigated peripheral blood (PB) and Bone Marrow (BM) lymphocyte subpopulations in a series of previously untreated DLBCL and FL. Aim of the study was: i. to assess whether LMR alteration is linked to a distinct lymphocyte subset; ii. to correlate PB with BM cell subpopulations; iii. to compared DLBCL and FL with regards to PB and BM lymphocyte subpopulations. Methods: Overall, PB cells from 58 DLBCL and 27 FL at diagnosis were investigated; BM cell populations were analyzed in 77 DLBCL and 54 FL. None of the studied patients had BM involvement nor circulating lymphoma cells detectable by cytometry; in addition, none of the patients had been previously treated with immunosuppressive drugs. PB from 40 healthy subjects and BM samples from 69 subjects without primary BM dysfunctions were used as control. Multicolor flow cytometry (MFC) was used to evaluate percentages and absolute values of the following cell subsets: CD3+ T cell, CD3-CD56+ NK cells, CD19+CD20+ B cells. Among B cells, t he analysis included transitional (CD38+high/CD24+high/IgD+/CD27-), naive (CD38+/CD24+/IgD+/CD27-), memory IgD+ (CD38-/+CD24+/IgD+/CD27+), and memory IgD- (CD38-/CD24+high/IgD-/CD27+) subsets; B cell precursors were investigated as CD19+CD20-CD10+ and mature B cells as CD19+CD20+CD10-. Results: A significant reduction in median absolute lymphocyte count was seen in PB from lymphoma patients (median: 1,526/µL) compared to controls (median: 2,195/ µL). Moreover, absolute lymphocytes were more markedly reduced in DLBCL (median: 1,386 /µL) compared to FL (median: 1883/µl) (p<0.001). This resulted in a significantly lower median PB-LMR in DLBCL (median LMR: 3) compared to FL patients (median LMR: 5) (p=0.001). Among PB cell subpopulations, a marked reduction in circulating B cells was observed in lymphoma patients (median 96/µL) compared to controls (median 224/µL). Again, loss of circulating B-cells was more markedly pronounced in DLBCL (median: 81.5 cell/µL) compared to FL (median: 150 cell/µL) (p=0.013). Differences in circulating B-cells among DLBCL, FL and controls are shown in Figure 1A. At BM analysis, a reduction of B cell precursor (CD19+/CD20-/CD10+) could be documented only in DLBCL. A reduction in circulating T-lymphocytes in lymphoma patients compared to controls was detected as well, although at a lower extent compared to B-cells. No significant abnormalities in the amount of circulating NK-cells was seen in both DLBCL and FL patients. When these data were matched with treatment outcome, the reduction of circulating B cells proved to be associated with an unfavorable outcome in younger patients with DLBCL. Indeed, among DLBCL patients aged < 60 yrs, a poorer progression free survival was seen in those having less than 96 B-cells/µL compared to those with PB B-cells equal or above 96 B-cells/µl (Figure 1B). Conclusions: The study indicates that circulating B lymphocytes are significantly reduced in both DLBCL and FL compared to age-matched healthy controls; however, their loss is much more pronounced in DLBCL compared to FL. Indeed, abnormalities in B-cell progenitors are present in BM from DLCBL but not from FL. In addition, B-cell reduction may have an unfavorable impact in the long-term outcome of DLBCL, at least in younger patients. Lastly, both B and T-cell subsets are decreased in DLBCL and FL whereas circulating NK subset does not seem to be affected. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Lymphoma Microenvironment Signature Predicts Clinical Outcomes in Diffuse Large B-Cell Lymphoma Patients

Background: Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease, whose response to standard chemotherapy is variable. Stratification of DLBCL into GCB (germinal center B-cell)- or ABC (activated B-cell)-like type, so-called cell-of-origin (COO) classification, has been performed based on gene expression signatures and/or immunohistochemistry. Since COO classification may not necessarily retain prognostic power, a more accurate risk stratification model is warranted.Purpose: The goal of this study is to identify a gene expression signature(s) that accurately predicts clinical outcomes of DLBCL patients.Hypothesis: During the process of normal B-cell development in germinal center (GC), a variety of microenvironment cells, including T cells, fibroblasts, dendritic cells and macrophages, coordinately regulate clonal selection and proliferation of B-cells. Therefore, we hypothesized that the differences in the contexts of lymphoma microenvironment could affect clinical outcomes of DLBCL.Methods: Standard genome-wide gene expression analysis, including microarray and RNA-seq, can only measure abundant transcripts derived from lymphoma cells. In contrast, cells constituting lymphoma microenvironment are minor fractions in lymphoma tissues, and transcripts derived from these cells are expressed at low levels. To quantitatively measure those lowly-expressed transcripts, we employed the nCounter system, which enables an accurate measurement of rare and highly-fragmented transcripts from formalin-fixed paraffin-embedded (FFPE) tissues.Results: We first examined expression levels of 1900 genes related to immune-, cancer- and kinase-pathways using tissues from 30 DLBCL cases. We then identified a set of genes that were differentially expressed between cases exhibiting good prognosis (alive > 2 years without relapse) and those with poor prognosis (disease progression < 1 year). Strikingly, differentially-expressed genes were not B cell-related genes, but those related to microenvironment cells, including T cells, fibroblasts and macrophages. Furthermore, high levels of microenvironment-related genes dictated favorable prognosis. To identify subtypes of microenvironment cells, we next performed single-cell q-PCR assays (C1 system) and a multi-color immunofluorescence analysis of DLBCL tissues (Mantra system). We found that presence of GC microenvironment components such as follicular helper T cells, CD11c+ macrophages and FGFR1+ stromal cells was an accurate predictor for favorable prognosis. We defined the GC microenvironment signature based on the expression levels of ICOS, CD11c and FGFR1 mRNA using nCounter and established a novel scoring system. We then validated the system using an independent DLBCL cohort consisting of 170 cases. Our scoring system enabled precise prognostic stratification among patients treated with R-CHOP independently of IPI status and traditional COO-based prognostic models (Figure 1). Finally, to explore potential correlation between the microenvironment signature and mutational status of DLBCL cells, we performed target capture sequencing of 100 DLBCL cases using a clinically-validated sequencing platform (OncoPanel). Our interim analysis did not reveal a correlation between specific lymphoma mutations and the microenvironment signature, suggesting that GC microenvironment signature, which we have identified, predicts clinical outcomes independently from mutational status of DLBCL cells.Conclusions: We identified a GC microenvironment signature that accurately dictates clinical outcomes and established a novel nCounter-based scoring system, which may inform prognostic stratification and the selection of chemotherapeutic regimen of DLBCL. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

A Multicenter Retrospective Review of Outcomes of Diffuse Large B Cell Lymphoma (DLBCL) in the Elderly Treated with RCHOP in Australia

▪BackgroundThe outcome of DLBCL in the elderly is well described in clinical trials with reported 2-year OS of 60-80%. However less well reported are aspects of care of elderly DLBCL patients treated outside the trial context, including rates of completion of induction chemotherapy and detailed reasons for dose delays and reductions and their effect on outcomes including remission rates and progression free survival (PFS). Here we describe an Australian experience of management of elderly DLBCL patients treated at 3 large institutions.MethodsThis is a multi-center, retrospective analysis of consecutive patients aged >=60, with DLBCL, planned to receive rituximab-chemotherapy . The standard regimens included 3 cycles (early stage disease) or 6 cycles (advanced) RCHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, prednisolone) followed by two rituximab doses in fit patients; R-miniCHOP for unfit and R-CE(etoposide)OP for patients with cardiacdysfunction. Disease response was PET-based. Patient data including included baseline characteristics, histology, clinical parameters, comorbidities, treatment outcomes and toxicity were extracted from hospital medical records. The primary endpoint was PFS; secondary endpoints included overall survival (OS), overall response rate and toxicity and rates of dose delays, dose reductions, hospital admission and completion of planned treatment. Descriptive statistics were used in the analysis.Results227 patients were evaluated from 2012-2017. Table 1 shows baseline characteristics and treatment. Median age was 73 years (range 60-97). Major comorbidities were present in 126 (55%) patients, with ischaemic heart disease (24%), diabetes (21%) and renal impairment (15%) the most common. Impaired cardiac function (EF<55%) was present in 25 (11%) of cases. G-CSF was used in 57 (25%) patients for primary prophylaxis and 20 (9%) patients post febrile neutropenia. Dose reductions occurred in 114 patients (50%): 59 (26%) upfront, 85 (38%) from cycle 2 onwards and 30 (13%) requiring both. Fifty-four patients (24%) had more than one dose reduction during treatment and 40 (18%) had dose delays. Only 19 (8%) patients did not receive all planned cycles of R-chemotherapy. Hospital admission was required in 167 (74%) cases. The median number of admissions was 4 (range 0-16) with a median inpatient duration of 7 days (range 1-83). Sixteen patients were admitted to ICU and 36 had limitations place to their resuscitation orders due to admission related morbidity . Of the evaluable patients (one center) for toxicity (n=98) the most common (12, 5%) grade 3-4 non-haematological toxicity was hepatic (ALT>2xULN), see table 2. Complete remission (CR) on PET following completion of treatment was achieved in 161 cases (71%) and was not impacted by dose reduction/delay (CR of 71% in these cases). Radiotherapy was administered in 56 (25%) patients, predominantly for initial bulky disease (48%) or for residual disease (40%). Median follow-up was 33 months. Two year PFS 69% was (95% CI 63-76%), OS 78% (95% CI 73-84%), see figures 1 and 2 for survival curves. The impact of dose reduction, dose delay and major co-morbidities on survival are under assessment and will be presented at the meeting.DiscussionIn our cohort, DLBCL in the elderly presented frequently as advanced stage, with high rates of B symptoms and extranodal disease. Clinically-relevant comorbidities were present in the majority of patients. Despite >90% of patients completing the planned number of treatment cycles, toxicity was significant with a high rate of hospital admission and dose reductions. In this real world setting, CR rates and survival were reassuringly comparable with published trial data. Elderly DLBCL patients with significant comorbidities have similar survival to the published data but close monitoring and early intervention for toxicity is needed. [Display omitted] DisclosuresHawkes:Roche: Other: Travel expenses; Bristol-Myers Squibb: Research Funding; Janssen: Membership on an entity's Board of Directors or advisory committees, Research Funding; Roche: Other: Travel expenses, Research Funding; Merck Sharpe Dohme: Research Funding; Takeda: Other: Travel expenses; Celgene: Research Funding; Merck Serono: Research Funding. Grigg:Takeda: Membership on an entity's Board of Directors or advisory committees; Bristol-Myers Squibb: Membership on an entity's Board of Directors or advisory committees; Amgen: Membership on an entity's Board of Directors or advisory committees, Other: Travel expenses; Merck Sharpe Dohme: Membership on an entity's Board of Directors or advisory committees; Roche: Membership on an entity's Board of Directors or advisory committees, Other: Travel expenses. Cheah:BMS: Membership on an entity's Board of Directors or advisory committees, Other: Conference travel expenses; Janssen: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Takeda: Speakers Bureau; Gilead: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Roche: Research Funding, Speakers Bureau; Celgene: Research Funding; Abbvie: Research Funding; Amgen: Other: Conference travel expenses.

Outcome of Diffuse Large B-Cell Lymphoma with First-line Chemotherapy

Background: Diffuse Large B-Cell Lymphoma (DLBCL), most common Non-Hodgkin Lymphoma (NHL) variety, is an aggressive, fast-growing form comprising up to 40% of all cases globally. Objective: To observe the treatment outcome of different subtypes of Diffuse Large B-Cell Lymphoma (DLBCL) after first-line chemotherapy and also the association with IHC, presenting age, sex, and IPI score with outcome. Methodology: This is a retrospective data analysis included all DLBCL patients registered in the department of Haematology of National Institute of Cancer Research and Hospital (NICRH) between July 2016 to June 2019. Results: Total 188 cases were included in this study and mean age was 48 years with a Standard deviation of 15 years with Male (69.1%) predominance. We divide the cases into three different entities of DLBCL [Germinal Centre B-cell like (GCB), Non-GCB and others (NOS) among them Non-GCB variety was the prevalent (47.3%) one. After first line chemotherapy 52.1% complete remission with 7% death was observed in overall outcome. There was no significant difference in outcome among different types of DLBCL after chemotherapy based on Han’s algorithm. Rituximab with CHOP has significantly better outcome than CHOP alone arm (p: 0.021). Conclusion: This limited database study of NICRH will help to ascertain the outcome of DLBCL after first-line chemotherapy in Bangladesh.