Subgroup Of Diffuse Large B-cell Lymphoma Research Articles

Correlation between PD-1 and sPD-L1 expression levels in peripheral blood of DLBCL patients and their clinicopathological characteristics.

Molecular pathology and clinical characteristics play a crucial role in guiding treatment selection and predicting the prognosis of diffuse large B-cell lymphoma (DLBCL). The programmed cell death protein 1 (PD-1) and its ligand (PD-L1), have emerged as pivotal regulators of immune checkpoints in cancer. The objectives of this study are to investigate the correlation between the expression levels of PD-1 and soluble PD-L1 (sPD-L1) in the peripheral blood of DLBCL patients, analyze their clinicopathological characteristics, and identify the optimal beneficiary group for PD-1/PD-L1 blockade. Peripheral blood samples were collected from 36 DLBCL patients before their initial treatment at Shandong Cancer Hospital between December 2018 and July 2019. The expression levels of PD-1 and sPD-L1 were measured using flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively. The clinicopathological characteristics, including age, sex, Ann Arbor stage, International Prognostic Index (IPI) score, response to treatment, etc., were recorded for each patient. The surface expression of PD-1 on peripheral blood T cells was significantly higher in DLBCL patients compared to healthy controls. There was a significant association between elevated PD-1 expression levels and the advanced Ann Arbor stage (P=0.0153) as well as the B group (P=0.0184). Higher sPD-L1 levels were associated with the GCB subtype according to Hans's classification (P=0.0435). The expression levels of PD-1 and sPD-L1 in the peripheral blood of DLBCL patients are significantly correlated with advanced disease stage, B group, and GCB subtype according to Hans's classification. This suggests that the PD-1/PD-L1 axis play a critical role in specific subgroups of DLBCL. Targeting this axis could serve as a potential therapeutic strategy to enhance the clinical outcomes of DLBCL patients. Further studies are necessary to explore the prognostic implications of PD-1 and sPD-L1 expression levels in DLBCL patients.

Mouse models of diffuse large B cell lymphoma.

Diffuse large B cell lymphoma (DLBCL) is a genetically highly heterogeneous disease. Yet, to date, the vast majority of patients receive standardized frontline chemo-immune-therapy consisting of an anthracycline backbone. Using these regimens, approximately 65% of patients can be cured, whereas the remaining 35% of patients will face relapsed or refractory disease, which, even in the era of CAR-T cells, is difficult to treat. To systematically tackle this high medical need, it is important to design, generate and deploy suitable in vivo model systems that capture disease biology, heterogeneity and drug response. Recently published, large comprehensive genomic characterization studies, which defined molecular sub-groups of DLBCL, provide an ideal framework for the generation of autochthonous mouse models, as well as an ideal benchmark for cell line-derived or patient-derived mouse models of DLBCL. Here we discuss the current state of the art in the field of mouse modelling of human DLBCL, with a particular focus on disease biology and genetically defined molecular vulnerabilities, as well as potential targeting strategies.

A Phase I Study of Copanlisib and Venetoclax in Patients with Relapsed or Refractory Diffuse Large B-Cell Lymphoma

Introduction: Diffuse large B-cell lymphoma (DLBCL) is a genomically heterogenous disease, which can be classified into distinct subgroups or clusters (Chapuy et al., Nat. Medicine, 2018, Wright et al., Cancer Cell, 2020). Leveraging knowledge of genetic alterations to design novel personalized treatment strategies may improve outcomes. Preclinical data suggest that the pan-phosphoinositide 3-kinase (PI3K) inhibitor, copanlisib, plus the BCL-2 inhibitor, venetoclax, are synergistic in genetically defined, high-risk, subtypes of DLBCL (Bojarczuk et al., Blood, 2019). We therefore hypothesized that copanlisib plus venetoclax would be an active treatment for patients with relapsed/refractory DLBCL, particularly in those with perturbed B-cell receptor (BCR)/PI3K signaling, which were predicted to be enriched in the relapsed setting. Here, we report the results of a phase I study of copanlisib plus venetoclax for patients (pts) with R/R DLBCL. Methods: This is an investigator-initiated, phase I, multicenter trial (NCT04572763) with the primary objective of determining the maximum tolerated dose (MTD) and recommended phase 2 dose (RP2D) of copanlisib and venetoclax in pts with R/R DLBCL. A standard 3+3 dose escalation design was employed, in which pts were treated with 60 mg IV copanlisib on days 1, 8, and 15 in addition to escalating doses of daily venetoclax for each 28-day cycle. During cycle 1, a venetoclax dose ramp-up was performed in the outpatient setting to a target dose of 200 mg (dose level (DL) +1), 400 mg (DL +2) and 800 mg (DL +3). The DLT observation period was the first cycle. Patients with progressive disease prior to the end of the DLT window were replaced. Key eligibility criteria included a confirmed diagnosis of DLBCL or high-grade B-cell lymphoma (HGBCL), relapse after or not a candidate for autologous stem cell transplant or chimeric antigen receptor (CAR) T-cell therapy, and adequate hematologic and organ function. Key exclusion criteria were poorly controlled hypertension or diabetes. CTCAE v5 and Lugano criteria were used to evaluate toxicity and efficacy, respectively. Results: As of July 21, 2023, 12 pts were evaluable. Patients were treated in DL +1 (n=5), DL +2 (n=3), and DL +3 (n=4). The median age was 62 years (range 24-87). Eighty three percent were male. Median number of prior treatments was 6 (range 1-11) and 10 pts had received prior CAR T-cell therapy. No DLTs were observed, though 2 pts in DL +3 were not able to escalate to 800 mg due to recurrent neutropenia and noncompliance, respectively. The most common toxicities were neutropenia (50%, all gr3/4), thrombocytopenia (42%, 8% ≥ gr3), nausea (33%, 8% ≥ gr3), fatigue (25%, 8% ≥ gr3), and headache (25%, 8% ≥ gr3) (Figure). Hyperglycemia occurred in 2 pts (both gr1). Serious adverse events occurred in 50% of pts and included fracture (n=1), hypoxia (n=2), and death (n=3), which were not related to study treatment. The deaths were related to progressive disease. Fifty percent of pts required drug holds (n=5 for venetoclax, n=5 for copanlisib). Sixty percent required growth factor support. There were no cases of laboratory or clinical tumor lysis syndrome. Sponsor support was withdrawn prior to completion of enrollment to DL+3, but DL+2 (copanlisib plus 400 mg venetoclax) was safe and well tolerated. The overall response rate (ORR) and complete metabolic response (CMR) rate was 8% (90% CI: 0 - 34%), as 1 pt in DL +1, who had received >5 lines of prior therapy, achieved a CMR, with response lasting 3 months. The median follow-up was 9.1 months. The median overall survival (OS) and progression-free survival (PFS) were 3.5 and 1.8 months, respectively. Conclusions: Copanlisib plus venetoclax is feasible and safe in pts with R/R DLBCL, with no DLTs observed. While it is possible that copanlisib plus venetoclax is active in select genomic subgroups of DLBCL, there was limited activity in this unselected, heavily pre-treated patient population.

Circulating Cell-Free DNA (ccfDNA) Concentrations Measured By Capillary Electrophoresis: High Cost-Effectiveness in Discriminating DLBCL Patients and Associations with High Tumor Burden

Introduction: Diffuse large B-cell lymphoma (DLBCL) is an aggressive malignancy with marked clinical-biological and prognostic heterogeneity. Its different clinical outcomes may be predicted by the identification of molecular biomarkers. In this sense, quantification of circulating cell-free DNA (ccfDNA), and mainly circulating tumor DNA (ctDNA) have emerged as minimally invasive methods able to predict its prognosis. Furthermore, ctDNA kinetics have been identified as a highly sensitive and specific marker for early detection of sensitivity and resistance to therapy, being associated with response rates, survival and prediction of relapses. However, such measurements are based on the use of costly technologies and with restricted access in clinical practice. Therefore, we aimed to quantify ccfDNA in DLBCL patients using capillary electrophoresis (CEF), a low-cost and widely available methodology, in order to establish prognostic associations between its serum concentrations ([ ]) and clinical-laboratory characteristics, as well as outcomes. Methods: This prospective and single-center study involved 54 DLBCL patients, diagnosed and treated at the University of São Paulo, from 2019 to 2020, and 23 healthy-controls. All cases were treated with 6-8 cycles of R-CHOP. Plasma samples were collected from DLBCL patients at three moments: at diagnosis (A), after the 4th CT cycle (B) and at the end of treatment (C). ccfDNA was extracted using the QIAamp MinElute ccfDNA© kit and quantified by CEF ( Agilent 2100 Bioanalyzer©). The ccfDNA quantification was established adding up the concentrations of DNA fragments between 75-200 bp. Comparison between two groups was performed using the Mann-Whitney test, and between three groups using the Kruskall-Wallis test. The cut-off for ccfDNA [ ] (1147 ng/µl) was determined by ROC methodology, with sensitivity = 71% and specificity = 78%. Survival curves were constructed using the Kaplan-Meier method and Log-Rank test was used to assess the relationship between variables and outcomes, and a p-value ≤ 0.05 was considered statistically significant. Results: The median age at diagnosis was 57 years (21-91) and 61.1% (33/54) were female. Advanced-stage (III/IV) was observed in 59.3% (32/54), 29.6% (16/54) had ECOG ≥ 2, 52.6% (23/54) had IPI ≥ 3, and 44.4% (24/54) had bulky ≥ 10 cm. BM infiltration and involvement of ≥ 2 EN sites occurred in 5.6% (3/54) and 16.8% (9/54), respectively. Median LDH was 308 U/L (138-2881). With a median follow-up of 32 months, the medians OS and EFS were 32 months (95% CI: 28-35) and 31 months (95% CI: 28-34), respectively. The ORR was 76% (95% CI: 63.5-86). The overall mortality rate was 20.4% (95% CI: 11.1-32.3). ccfDNA [ ] were significantly higher in DLBCL patients compared to controls (p<0.001), however, it did not differ significantly between samples from DLBCL patients collected at the 3 different time points (p=0.602) Figure 1. We observed positive associations between ccfDNA [ ] in diagnostic samples of DLBCL patients and biomarkers of high tumor burden, including higher LDH levels (p=0.003), bulky (p=0.011), ECOG ≥ 2 (p<0.001), and IPI ≥ 3 (p=0.069) Figure 2. However, ccfDNA [ ] at diagnosis were not statistically associated with ORR (p=0.264) Figure 2, nor with clinical outcomes, including OS (1-year OS: 92% for [ccfDNA] < 1147 ng/µl vs 76% for [ccfDNA] ≥ 1147 ng/µl, p=0.217) and EFS (1-year EFS: 92% for [ccfDNA] < 1147 ng/µl vs 69% for [ccfDNA] ≥ 1147 ng/µl, p=0.270) Figure 3. We strongly believe that the short FUP and sample number restriction may have contributed to the lack of statistically significant association between ccfDNA [ ] and responses/outcomes observed. In this sense, cohort expansion and NGS for ctDNA identification are ongoing to assess its potential impact as a prognostic biomarker in our cohort. Conclusion: Here we demonstrated that ccfDNA [ ] measured by CEF was able to accurately discriminate healthy individuals from DLBCL, highlighting its potential role as a minimally invasive diagnostic biomarker. Furthermore, ccfDNA [ ] at diagnosis in DLBCL patients was positively associated with markers of high tumor burden and adverse prognosis, such as higher LDH levels, bulky disease, poor PS and high-risk IPI. Our results suggest that measurement of ccfDNA by CEF may be a highly cost-effective methodology to identify high-grade B-cell NHLs and recognize DLBCL subgroups with adverse clinical-prognostic characteristics.

Loncastuximab Tesirine Demonstrated Substantial Single-Agent Efficacy and Manageable Safety Profile in Heavily Pretreated Chinese Patients with Relapsed or Refractory (R/R) Diffuse Large B-Cell Lymphoma (DLBCL)

Introduction: Patients with R/R DLBCL who failed multi-agent chemoimmunotherapies usually have a poor prognosis, leaving a significant unmet medical need. Loncastuximab tesirine (Lonca) is an antibody-drug conjugate, composed of a humanized anti-CD19 antibody conjugated to a pyrrolobenzodiazepine dimer toxin that showed clinical efficacy and manageable safety in heavily pretreated DLBCL patients in a global phase 2 study of Lonca monotherapy (NCT03075696). Here we present results from a phase 2 study (ChiCTR2300072058) of Lonca monotherapy in Chinese patients with R/R DLBCL for the first time, including analyses of responses in high-risk DLBCL subgroups. Methods: This open-label, single-arm, phase 2 study was designed to confirm the safety, efficacy and PK profiles of Lonca monotherapy in Chinese patients with R/R DLBCL who had failed ≥ 2 lines of systemic therapies. The primary endpoint was overall response rate (ORR) as assessed by IRC according to the Lugano 2014 criteria. Secondary efficacy endpoints include complete response rate (CRR), DOR, RFS, PFS, and OS. Treatment-emergent adverse events (TEAEs) were reported by CTCAE v4.0. Results: As of data cutoff (11-Jan-2023), 64 Chinese patients with DLBCL were enrolled and received at least one dose of Lonca (median: 4.0 cycles [range: 1-17]) and were evaluable for safety and efficacy analyses. The median number of prior lines of therapies was 3.0 (range: 2-12), with 67.2% of patients having ≥ 3 lines of prior therapies. Four patients (6.3%) had received prior CD19 CAR-T therapy. Forty-three patients (67.2%) were refractory to the first-line therapy (primary refractory), 56 patients (87.5%) were refractory to the most recent line of therapy and 40 patients (62.5%) were refractory to all prior lines of therapies. At data cutoff, the median follow-up was 8.5 months. The ORR by IRC was 51.6% (95% CI: 38.7% to 64.2%), and CRR was 23.4%. The median DOR (mDOR) was 6.37 months as assessed either by IRC or by investigator. For patients with a CR, the mDOR was 10.22 months (6.08 months for patients with PR). The median time to first response was 41.0 days. The median PFS and median OS were 4.96 and 9.33 months, respectively. Responses, including CRs, were observed in several high-risk subgroups (Fig. 1): ORR was 54.7% in patients with advanced stage disease (Stage III/IV) and 50.0% in patients with transformed disease. Patients who were refractory to first-line therapy, to the most recent therapy, or to all prior therapies had ORRs of 44.2%, 51.8% and 42.5%, respectively. Three patients who had received a prior SCT had an ORR of 100%. Lonca was also effective in elderly patients ≥ 65 years (ORR of 45.8%, mDOR of 6.37 months) and in patients who had received prior CD19 CAR-T therapy (ORR of 50%). Overall, 64 (100%) patients had at least one TEAE and 61 (95.3%) patients had at least one TEAE ≥ Grade 3. The most common (≥30%) all-grade TEAEs were GGT increased (71.9%), followed by anaemia (70.3%), platelet count decreased (65.6%), AST increased (64.1%), WBC count decreased (64.1%), neutrophil count decreased (60.9%), ALT increased (51.6%), hypokalaemia (37.5%), and blood ALP increased (32.8%). The most common (≥15%) TEAEs of Grade ≥3 were platelet count decreased (34.4%), neutrophil count decreased (28.1%), WBC count decreased (28.1%), GGT increased (25.0%), anaemia (18.8%), hypokalaemia (18.8%), neutropenia (17.2%), and lymphocyte count decreased (15.6%). The rate of febrile neutropenia was low (3.1%). The majority of Grade ≥ 3 TEAEs were abnormalities of clinical laboratory values as opposed to clinical symptoms and were generally reversible. Treatment-related TEAEs leading to treatment discontinuation occurred in 10 patients (15.6%), most commonly GGT increased (3 patients; 4.7%). No increase in toxicity was seen in patients aged ≥65 years compared with younger patients. Conclusions: Lonca demonstrated substantial and clinically meaningful single-agent efficacy and was well-tolerated in heavily pretreated Chinese patients with R/R DLBCL. Toxicities were generally manageable and reversible in most patients by routine clinical practice and/or with dose modifications, with neither unexpected safety concerns nor different toxicity profile in patients aged ≥65 years. Encouraging and durable responses were also observed in high-risk patient groups, including patients with advanced stage, transformed or refractory DLBCL.

Depth of metabolic response at interim PET and survival outcomes among patients with primary refractory and early relapsing diffuse large B-cell lymphoma (DLBCL).

7543 Background: Total metabolic tumor volume (TMTV) and maximum standardized uptake value (SUVmax) may improve upon standard 18F-fluorodeoxyglucose positron emission tomography/computed tomography (PET-CT)-based assessment of advanced DLBCL during treatment. We report the association of TMTV and SUVmax changes at interim PET-CT (PET2) on survival outcomes in patients with primary refractory DLBCL (prDLBCL) and early relapsing DLBCL (erDLBCL). Methods: Adult patients with prDLBCL (less than a complete response (CR) or progression by the end of treatment (EOT)) or erDLBCL (CR at EOT with relapse within 12 months) between 2005 and 2019 at Mayo Clinic Rochester were included. The % change (Δ) from baseline to PET2 (after 2 cycles of 1L therapy) of TMTV and SUVmax were measured. A PET segmentation threshold of 1.5 liver mean SUV + two standard deviations with a minimum volume constraint of 0.5 mL was utilized (MIM Software, Inc., Cleveland OH, USA) with manual input as needed. Functional splines analysis determined SUVmax and TMTV threshold for analysis. Results: 131 patients had complete PET-CT data (N=79 prDLBCL, N=52 erDLBCL). Baseline median TMTV and SUVmax were 767.4 cm3 (range 0.9-6840.4) and 20.5 (2.0-52.6), respectively. At a median follow up of 77.6 months, 81 patients had died. 2-year OS rate was 53% (95% CI: 42-67) for patients with a PET2 ΔSUVmax decline ≥65% compared to 15% (95% CI: 7-29) for patients with a PET2 ΔSUVmax decline <65%(p<0.001). PrDLBCL had a 2-year OS rate of 48% (95% CI: 31-74) for a ΔSUVmax ≥65% compared to 7% (95% CI: 2-21) for a ΔSUVmax <65%(p<0.001), which captured 58% of prDLBCL patients (N=46). Two-year OS rate was 43% (95% CI: 34-54) for patients with a ΔTMTV ≥75% compared to 5% (95% CI: 1-35) for patients with ΔTMTV <75% (p<0.001). All patients with a ΔTMTV <75% had prDLBCL. PrDLBCL had a 2-year OS rate of 28% (95% CI: 18-44) for a ΔTMTV ≥75%. The outcomes for ΔSUVmax <65% and ΔTMTV <75% at PET2 remained significant in a cox regression model adjusted for IPI and bulky disease (>10cm)(Table). Conclusions: A ΔTMTV decline <75% or a ΔSUVmax decline <65% at PET2 identified an ultra-high-risk subgroup of DLBCL with particularly poor outcomes, that captured a high percentage of prDLBCL patients, and was significant after adjusting for IPI and bulky baseline disease. These patients may benefit from clinical trials evaluating alternative PET-adapted treatment strategies, but further evaluation among all patients with DLBCL is needed. [Table: see text]

17th International Conference on Malignant Lymphoma, Palazzo dei Congressi, Lugano, Switzerland, 13 - 17 June, 2023.

Background: Previous researches have mainly focused on the discussion of whether cancer stem cells exist in diffuse large B-cell lymphoma (DLBCL). However, some subgroups with dismal prognosis that exhibit stem cell-like characteristics have been overlooked. Methods: Using an innovative one-class logistic regression (OCLR) machine learning algorithm, we calculated stemness indices and systematically assessed their ability to reflect the oncogenic dedifferentiation-like characteristics of DLBCL. Next, we identified signatures associated with oncogenic dedifferentiation-like features and prognosis using LASSO and SVM-RFE algorithms and developed a proportional hazards model (Riskscore) for DLBCL through Cox regression. Then we stratified the risk of DLBCL (n = 2133) and evaluated the prognostic value of the risk model across known clinical and molecular subgroups. We further compared the characteristics of high- and low-risk DLBCL with Burkitt lymphoma. Finally, we investigated the mechanisms of poor prognosis in high-risk DLBCL using transcriptomics, genomics (n = 576) and single-cell RNA sequencing (n = 19) data, as well as cell experiments and internal validation cohorts. Results: In this study, we identified and validated a DLBCL subgroup (25.6% of DLBCL) with stem cell-like characteristics and dismal prognosis. This high-risk group was defined as polyamine metabolism-cold immune tumor with upregulated polyamine metabolic key enzyme (ODC1) and desert-like immune infiltration, and had a poor prognosis with lower 3-year OS rate (54.3% vs. 83.6%, p < 0.0001) and PFS rate (42.8% vs. 75.2%, p < 0.0001) compared to the low-risk group. We found that some patients with MYC rearrangement, double-hit, double-expresser, or complete remission may have either favorable or poor prognosis, which can be accurately identified by our risk stratification model. Additionally, the high-risk group exhibited malignant proliferative phenotypes similar to Burkitt lymphoma. Genomic analysis revealed widespread copy number losses in the chemokine and interferon coding regions 8p23.1 and 9p21.3 in the high-risk group. Bulk and single-cell transcriptome analysis indicated that the upregulation of ODC1 might mediate the cold immune microenvironment of DLBCL, and knocking down ODC1 effectively inhibited DLBCL cell proliferation. Promisingly, our model effectively identified patients who were insensitive to immunotherapy (CAR-T and immune checkpoint antibodies). The research was funded by: This work was supported by grants from the National Natural Science Foundation of China (grant no. 81873450, 82170181), Beijing Hospitals Authority Youth Programme (code: QML20200201), and Beijing Natural Science Foundation (No. 7222027) to Liang Wang. Keywords: Bioinformatics, Computational and Systems Biology, Microenvironment, Risk Models No conflicts of interests pertinent to the abstract.

Identification and validation of hub genes in CD5-positive diffuse large B-cell lymphoma.

CD5+ diffuse large B-cell lymphoma (DLBCL), as a significant heterogeneity category of DLBCL, is reflected in both the molecular biological and genetic levels, which in turn induces ever-changing clinical manifestations, and what mediates tumor survival mechanisms are still unclear. This study aimed to predict the potential hub genes in CD5+ DLBCL. A total of 622 patients with DLBCL diagnosed between 2005 and 2019 were included. High expression of CD5 was correlated with IPI, LDH, and Ann Arbor stage, patients with CD5-DLBCL have longer overall survival. We identified 976 DEGs between CD5-negative and positive DLBCL patients in the GEO database and performed GO and KEGG enrichment analysis. After intersecting the genes obtained through the Cytohubba and MCODE, further external verification was performed in the TCGA database. Three hub genes were screened: VSTM2B, GRIA3, and CCND2, of which CCND2 were mainly involved in cell cycle regulation and JAK-STAT signaling pathways. Analysis of clinical samples showed that the expression of CCND2 was found to be correlated with CD5 (p = 0.001), and patients with overexpression of CCND2 in CD5+ DLBCL had poor prognosis (p = 0.0455). Cox risk regression analysis showed that, for DLBCL, CD5, and CCND2 double positive was an independent poor prognostic factor (HR: 2.545; 95% CI: 1.072-6.043; p = 0.034). These findings demonstrate that CD5 and CCND2 double-positive tumors should be stratified into specific subgroups of DLBCL with poor prognosis. CD5 may regulate CCND2 through JAK-STAT signaling pathways, mediating tumor survival. This study provides independent adverse prognostic factors for risk assessment and treatment strategies for newly diagnosed DLBCL.

Molecular classification and therapeutics in diffuse large B-cell lymphoma.

Diffuse large B-cell lymphoma (DLBCL) encompasses a wide variety of disease states that have to date been subgrouped and characterized based on immunohistochemical methods, which provide limited prognostic value to clinicians and no alteration in treatment regimen. The addition of rituximab to CHOP therapy was the last leap forward in terms of treatment, but regimens currently follow a standardized course when disease becomes refractory with no individualization based on genotype. Research groups are tentatively proposing new strategies for categorizing DLBCL based on genetic abnormalities that are frequently found together to better predict disease course following dysregulation of specific pathways and to deliver targeted treatment. Novel algorithms in combination with next-generation sequencing techniques have identified between 4 and 7 subgroups of DLBCL, depending on the research team, with potentially significant and actionable genetic alterations. Various drugs aimed at pathways including BCR signaling, NF-κB dysfunction, and epigenetic regulation have shown promise in their respective groups and may show initial utility as second or third line therapies to patients with recurrent DLBCL. Implementation of subgroups will allow collection of necessary data to determine which groups are significant, which treatments may be indicated, and will provide better insight to clinicians and patients on specific disease course.

Advanced Ann Arbor stage and age over 60 years as prognostic predictors in patients with primary cervical lymphoma: a retrospective cohort study and systematic review

ObjectiveTo evaluate the overall survival (OS), disease-specific survival (DSS), and recurrence-free survival (RFS) for primary cervical lymphoma (PCL), an extremely rare disease without treatment consensus.MethodsWe conducted a retrospective study included 177 patients, including 169 cases identified from literature review. The Kaplan-Meier methods and Cox regression were used to determine the OS, DSS, RFS, and relevant risk factors.ResultsThe 5-year OS and 5-year DSS rates were 85.8 and 87.2%, respectively, while the 5-year RFS rate was 85.5%. Diffuse large B-cell lymphoma (DLBCL) was the predominant subtype that comprised 63.8% (113 cases) in this cohort. Multivariate analysis in the DLBCL subgroup revealed that age ≥ 60 years (Odds ratio [OR]: 26.324, 95% Confidence Interval [CI]: 5.090–136.144, P < 0.001) or stage IIIE-IVE (advanced stage) (OR: 4.219, 95%CI: 1.314–13.551, P = 0.016) were the risk factors for OS, while patients with age ≥ 60 years (OR:23.015, 95%CI: 3.857–137.324, P = 0.001), and stage IIIE-IVE (OR: 4.056, 95% CI: 1.137–14.469, P = 0.031) suffered a poor DSS. Chemotherapy and/or radiotherapy improved the OS (P = 0.008), DSS (P = 0.049), and RFS (P = 0.003). However, cancer-directed surgery did not improve the OS, DSS, and RFS. The risk factor was unavailable in other subtypes of PCL due to limited cases.ConclusionThe survival outcomes in patients with PCL at early stage were satisfactory, while the advanced disease stage and age ≥ 60 years were the two major factors predicting poor prognosis in DLBCL subtype.

Real-World Outcomes in Patients with Relapsed or Refractory Diffuse Large B-Cell Lymphoma Treated with Standard of Care: A Cota Database Analysis

Introduction: Despite the rapidly evolving therapeutic landscape for relapsed or refractory (R/R) diffuse large B-cell lymphoma (DLBCL), treatment with chemotherapy, single-agent rituximab/obinutuzumab, single-agent lenalidomide, or combinations of these agents are common components of R/R DLBCL treatment regimens. This study aimed to characterize clinical outcomes in a real-world cohort of patients with R/R DLBCL treated with the above SOC regimens in the second-line or later setting and to assess the impact of patient demographics and clinical characteristics on clinical outcomes. Methods: This retrospective, observational study used data from the COTA electronic health records database (COTA, New York, NY). The study included adults diagnosed with DLBCL between January 1, 2010, and March 31, 2022, treated with ≥ 1 prior systemic line of therapy (LOT), including ≥ 1 anti-CD20 monoclonal antibody-containing regimen. Patients had an evaluable response assessment for these R/R DLBCL treatment regimens: chemotherapy, single-agent rituximab/obinutuzumab, single-agent lenalidomide, or combinations of these agents. Treatment outcomes were evaluated using real-world overall response rate (rwORR), real-world complete response (rwCR) rate, real-world duration of response (rwDOR), real-world duration of complete response (rwDOCR) rate, real-world progression-free survival (rwPFS), and overall survival. Subgroup analyses were conducted for patients with ≥ 2 prior LOTs. Logistic regression and Cox proportional hazards regression analyses evaluated the association between outcomes and patient demographics and clinical characteristics. Results: Overall, 573 patients with R/R DLBCL who had ≥ 1 prior LOT were included in the study; of these, 179 patients (31.2%) had ≥ 2 prior LOTs. Median duration of follow-up was 7.7 months. More than one-half (57.1%) were men; 49.9% were aged > 65 years (aged > 75 years, 20.8%) at diagnosis; 82.0% had an ECOG performance status of 0 to 1; 2.8% had prior exposure to chimeric antigen receptor T-cell (CAR T) therapy; 7.0% had prior autologous stem cell transplant; 62.0% had primary refractory disease; 68.6% were refractory to their last LOT; and 67.7% were refractory to their most recent anti-CD20 therapy. Patients experienced worsening outcomes as they progressed through LOTs. In patients with ≥ 1 prior LOT, rwORR was 52% (95% CI: 48-56), rwCR rate was 23% (95% CI: 19-27), median rwPFS was 3.0 (95% CI: 2.8-3.3) months, and median overall survival (mOS) was 12.9 (95% CI: 10.1-16.9) months. In the subgroup with ≥ 2 prior LOTs, rwORR was 40% (95% CI: 33-48), rwCR rate was 13% (95% CI: 8-19), median rwPFS was 2.4 (95% CI: 2.0-2.7) months, and mOS was 6.2 (95% CI: 5.2-10.9) months. In patients with ≥ 3 prior LOTs, rwORR was 34% (95% CI: 23-45), rwCR rate was 8% (95% CI: 3-16), rwDOR was 2.3 (95% CI: 1.3-3.3) months, rwDOCR was 8.5 (95% CI: 2.1-not estimable) months, median rwPFS was 1.9 (95% CI: 1.5-2.5) months, and mOS was 5.1 (95% CI: 3.2-7.3) months. Factors consistently associated with worse efficacy outcomes were primary refractory disease, refractory to last LOT, and refractory to last anti-CD20 therapy (Table). These characteristics indicated elements of a patient population with difficult-to-treat R/R DLBCL, with a 48%-66% lower likelihood of achieving a rwORR; a 45%-101% higher risk of progression; and a 39%-111% higher risk of mortality than those without these characteristics. Discussion: Patients with R/R DLBCL who received an increasing number of prior LOTs, had primary refractory disease, were refractory to their last LOT, were refractory to their last anti-CD20 therapy, or had prior exposure to CAR T therapy had poor outcomes, with an rwORR (rwCR rate) of 40% (13%) and 34% (8%) in patients with ≥ 2 prior LOTs and ≥ 3 prior LOTs, respectively. This is consistent with prior retrospective analyses of pretreated DLBCL (Crump et al, Blood 2017; Hamadani et al, Clin Lymphoma Myeloma Leuk 2022). Conclusions: Outcomes in patients with R/R DLBCL treated with chemotherapy, single-agent rituximab/obinutuzumab, single-agent lenalidomide, or combinations of these agents remain poor, especially those with difficult-to-treat R/R DLBCL. These findings underscore the high unmet need for new, safe, and effective therapies across this population, including the difficult-to-treat R/R DLBCL subgroups. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Genome-Wide Methylation and Haplotype-Resolved Aberrant Somatic Hypermutation Patterns in B-Cell Lymphomas

Introduction Diffuse large B-cell lymphoma (DLBCL) is a genetically, phenotypically, and clinically heterogenous disease. Multiple molecular classification systems using either genomic aberrations or gene expression profiles are used to delineate subgroups with shared biology. The cell of origin (COO) gene expression-based classification system stratifies DLBCLs into germinal center B-cell-like (GCB) and activated B-cell-like subgroups of DLBCL. More recently, the double-hit signature (DHITsig) has been used to further stratify GCB DLBCLs into DHITsig+ and DHITsig-, where the former is characterized by a gene expression profile reminiscent of the GC dark zone and inferior outcomes. Currently, patterns of somatic driver mutations can only partially explain the expression of DHITsig in some tumors. Genetic features such as DNA methylation have been shown to be associated with transcriptional repression and altered chromatin structure. We hypothesized that resolving DNA methylation patterns genome-wide using long read whole genome sequencing (LR-WGS) may reveal epigenetic differences associated with differential gene expression profiles within the DHITsig+ and DHITsig- subgroups and in the context of histologic transformation from follicular lymphoma. Methods Whole genome sequencing data of 10 patient samples and 4 cell line GCB DLBCL samples were generated using both short read (SR-WGS; Illumina) and LR-WGS (Oxford Nanopore Technologies, PromethION). The complementary approach combines somatic variant calling from highly accurate SR data with DNA methylation calling from LR data. GCB samples were selected based on the NanoString DLBCL90 assay and further subdivided into DHITsig+ (N=4) and DHITsig- (N=3). RNA sequencing data of 9 patient samples and 3 GCB DLBCL cell lines were generated and analyzed using Salmon and DESeq2. Simple somatic mutations from SR-WGS data were called using an ensemble of somatic variant callers, which confirmed all cases to belong to the EZB genetic subgroup. Methylated bases were called with Nanopolish followed by a differential methylation analysis using the DSS package in R to identify differences in methylation between both subgroups. Differentially methylated regions (DMRs) were annotated for GC B-cell specific superenhancers, CpG islands, and genes. Using WhatsHap and Nanomethphase, heterozygous single nucleotide polymorphisms (SNPs) from SR and LR data were used to phase LRs into haplotypes. Haplotype analyses were performed to determine the phase of aberrant somatic hypermutation (aSHM) and methylation patterns in the context of proximal translocation breakpoints. Results Most DMRs were found to be associated with regulatory regions such as promoters, superenhancers, and CpG islands. We found methylation differences between DHITsig+ and DHITsig- subgroups in 1430 superenhancers known to be active in GC B-cells. Within these superenhancer regions, DHITsig+ tumors were typically hypomethylated relative to DHITsig-. Several DHITsig classifier genes, including WNK2, TNFSF8, and IL21R, had methylation patterns inverted relative to the typical expression pattern (i.e. low methylation with high expression). Similarly, among 633 differentially methylated promoters, several including TCF3 and WNK2 demonstrated the same inverted methylation pattern. CpG islands showed distinct patterns of methylation in both subgroups that were inversely associated with expression, this also included genes differentially expressed between DHITsig+ and DHITsig- tumors. The DHITsig- group had increased levels of methylation of DHITsig genes in both promoters and gene bodies suggesting a functional role for gene body methylation. Finally, we observed that a high density of mutations near the BCL2 transcription start site consistently phased into the same haplotype as the BCL2-IGH translocation in five tumours, demonstrating the allele specificity of aSHM associated with these events (Panel B). Conclusions These results demonstrate differences in methylation between the subgroups. The association of hypomethylated superenhancers, promoters, and CpG islands with genes over-expressed in DHITsig+ suggests these methylation patterns may contribute to the DHITsig phenotype. These results may aid in improving GCB DLBCL patient classification and reveal factors contributing to its heterogeneity. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Combination of Phosphodiesterase 4 (PDE4) Inhibitor Roflumilast and R-CHOP As First-Line Therapy for High-Risk Diffuse Large B Cell Lymphoma Patients

BACKGROUND PDE4 inhibition is a biologically well-informed strategy with activity in B-cell malignancies including diffuse large B-cell lymphoma (DLBCL) (PMID: 15331441, PMID: 26503641). Roflumilast (R) is a PDE4 inhibitor, currently FDA-approved for COPD (PMID: 26338438 ). Building on preclinical and clinical activity of R in B-cell lymphomas, both alone and in combination with other agents (PMID: 27756749); a phase 1 evaluation was conducted, adding R to rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) in patients with untreated high risk DLBCL. METHODS: This is a single center, phase 1b, open label, non-randomized, single arm, investigator initiated trial (IIT) of R + R-CHOP (RR-CHOP) for treatment naïve high-risk DLBCL patients. Key eligibility criteria included: non-GCB DLBCL (Han's algorithm), NCCN-IPI risk score of ≥ 2, Ann Arbor stage II-IV; key exclusion criteria include active CNS involvement and documented history of severe depression (a specific concern related to the use of PDE4 inhibitors). Patients received R-CHOP every 21 days for 6 cycles and R PO daily (500 mcg), throughout the 18-week treatment period. Paired germline-tumoral exome sequence was completed for 70% of patients enrolled in the trial, and DLBCL genetic subtype defined by the probabilistic classification tool LymphGen (PMID: 32289277). Primary end point was assessment and grading of adverse events (AE) according to the CTCAE version 4.03 criteria. Eligible patients were included in the intent-to-treat analysis. Secondary end points were estimation of complete response (CR) as per Lugano criteria, progression free survival (PFS), overall survival (OS) and their relationship to DLBCL genetic subtypes. RESULTS: Ten patients met eligibility criteria for enrollment. Sixty percent were male, 50 % were Hispanics (HI), median age at diagnosis was 59 years old (y.o) (33 y.o-79 y.o), 50% were stage 2, and 50% were stage 3-4; 50% received CNS prophylaxis given high risk and 70% had International Prognostic Index (IPI) scores of 3 to 5. Standard doses of R-CHOP were able to be delivered safely, with addition of 500 mcg of R on daily basis. Regarding hematologic adverse events (AE); 1 patient (10%) experienced neutropenia grade 3, and two patients (20%) had neutropenia grade 2. Anemia occurred in 2 patients (20%), all grade 1-2. Regarding non-hematologic AE; anorexia occurred in 40% of the cohort, all grade1; diarrhea occurred in 50%, all grade 1; headache occurred in 60%, with 40% being grade 2, and 20% being grade 3, with the latter judged to be unrelated to RR-CHOP by the investigators. Forty percent had grade 1 increase of BNP; 40% of patients had nausea, 30% being grade 1-2, and 10% being grade 3; and 40% experienced weight loss, all grade 1-2. Of 10 enrolled patients, 9 were found evaluable for assessment of efficacy, with 66% achieving CR, 22% partial response and 11 % stable disease. At 50 months of follow up, PFS was 60% at 1 year (y) and 50% at 2 y. Median PFS was not reached. OS was 100% at 1 y and 86% at 2 y, with a median OS of 38.9 months. (Figure 1 and Figure 2). The recommended R dose for phase 2 is 500 mcg PO daily. LymphGen assigned the DLBCLs in this cohort to the A53, MCD, EZB and B2N subtypes. Notably, 3 of the 3 DLBCLs in our cohort classified as MCD or A53 DLBCLs, which normally predict poor outcome, reached CR and are long term responders. Examination of 461 primary DLBCLs (NCBI dbGaP Genomic Variation in DLBLC cohort) uncovered significantly higher PDE4B expression in in the MCD subtype, suggesting that addition of R may be particularly beneficial in this DLBCL subset. CONCLUSION: Phosphodiesterase Inhibition in DLBCL is a well biologically informed strategy with the potential to enhance and perhaps optimize the duration of responses to R-CHOP. On this difficult to treat subgroup of non-GC DLBCL, with known subpar outcomes, RR-CHOP demonstrated to be safe, with manageable AEs that did not differ from what we have historically seen with R-CHOP alone. Early efficacy data from this combination shows 10% improvement compared to our own institutional historical controls of treatment naïve non-GC DLBCL patients treated with R-CHOP. Based on this data, we are pursuing a phase 2 multisite IIT of RR-CHOP in patients with newly diagnosed high risk DLBCL. Despite the small size cohort, our data suggest that tailoring this strategy to the poor outcome MCD/A53 genetic subgroups may increase the likelihood of success. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Polyostotic DLBCL Is Characterized By a NF-Κb Pathway Affecting Molecular Profile and Superior Survival

Introduction Polyostotic diffuse large B-cell lymphoma (DLBCL) is a rare extranodal disease with only multifocal bone localizations that has not extensively been studied. Other known bone DLBCL comprises primary bone (PB-)DLBCL with a single bone localization with(out) locoregional lymphadenopathy and disseminated-DLBCL with ≥1 bone lesion(s) and ≥1 (extra) nodal localization(s). Recently (PMID: 34478526, de Groen et al.), we demonstrated a germinal center B-cell (GCB)-subtype with a centrocyte-like phenotype and a unique corresponding molecular profile, reflecting favorable prognosis for PB-DLBCL compared to non-bone DLBCL with a GCB-subtype. Here, the molecular characteristics of polyostotic-DLBCL were assessed and compared to PB-DLBCL, disseminated-DLBCL, and nodal DLBCL-GCB, using an extensive targeted next-generation sequencing (tNGS) panel. Methods This retrospective multicenter study analyzed 14 polyostotic-DLBCL, 44 PB-DLBCL, 26 disseminated-DLBCL, and 36 nodal DLBCL-GCB cases (selection based on PET/CT-scan), including well-annotated histological and clinical data. Cases were diagnosed with immunohistochemistry following the revised WHO classification of 2016. Cell-of-origin (COO) was determined by Hans' algorithm. Presence of MYC/BCL2/BCL6-rearrangements were identified through fluorescent in situ hybridization (FISH) and break-apart probes. High-grade B-cell lymphomas (double/triple hit) were excluded. As described (PMID: 34478526), tNGS was performed, using an extended panel consisting of 128 (LYMFv2, mostly used) or 52 (LYMFv1) B-cell lymphoma relevant genes. Results Our cohort consisted of 120 patients (78 males, 65%) with a median age at diagnosis of 61 (range 13-91). Half of the polyostotic-DLBCL cases belonged to a GCB phenotype (8/14, 57%), while a GCB phenotype was more frequently identified in PB-DLBCL (31/44, 70%) and disseminated-DLBCL (18/26, 69%). Using tNGS with LYMFv2 and LYMFv1, respectively 57 and 36 cases were successfully analyzed, elucidating (in)activating pathogenic mutations in 124 different genes. In polyostotic-DLBCL, frequent mutations in f.e.CARD11 and MYD88 demonstrated a unique mutational profile affecting the NF-κB pathway characteristic of ABC-genotypes (Figure 1, P=0.018), compared to PB-DLBCL, disseminated-DLBCL and nodal DLBCL-GCB. In contrast, an exclusive immunomodulatory profile with frequent mutations in B2M, IRF8 and TNFRSF14 was identified for PB-DLBCL (P=0.029). Focusing on GCB-associated epigenetic genes (CREBBP, EP300, EZH2, KMT2D, etc.), no significant difference was found between the mentioned DLBCL subgroups. Mutational profiles of disseminated-DLBCL revealed a broad spectrum of abnormalities (BCL2, CREBBP, TNFRSF14, and TP53), partially consistent with either an immunomodulatory or an epigenic genotype. Nodal DLBCL-GCB was specifically characterized by frequent aberrations in BCL2 and MYC and lacked an immunomodulatory or NF-κB activating genotype. Given >1 extranodal localizations, polyostotic-DLBCL is classified as high-risk disease with Ann Arbor stage IV. Compared to disseminated-DLBCL with similar stage III/IV disease, polyostotic-DLBCL demonstrated a superior 3-year overall survival (Figure 2, log-rank, p<0.001, HR: 4.76, CI: 1.4-15.9). Conclusion Polyostotic-DLBCL showed a unique mutational profile with frequent NF-κB-activating mutations, in contrast to PB-DLBCL, disseminated-DLBCL and nodal DLBCL-GCB. Despite this ABC-associated profile of polyostotic-DLBCL and its high Ann Arbor stage, a favorable prognosis was identified especially compared to similar high Ann Abor stage disseminated-DLBCL. To study COO and tumor microenvironment in more detail to evaluate polyostotic-DLBCL as a separate biological disease entity, gene-expression profiling with the BLYM777 panel (PMID: 35454765) will be presented at the meeting. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Molecular determinants of clinical outcomes in a real-world diffuse large B-cell lymphoma population

AbstractMolecular heterogeneity of diffuse large B-cell lymphoma (DLBCL) underlies the variable outcomes achieved with immunochemotherapy. However, outcomes of gene expression profiling (GEP)–defined molecular subgroups in a real-world DLBCL population remain unknown. Here we examined the prevalence and outcomes of molecular subgroups in an unselected population of 1149 patients with de novo DLBCL in British Columbia, Canada. Evaluable biopsies were profiled by fluorescence in situ hybridization (FISH), immunohistochemistry, and digital GEP to assign cell-of-origin and the so-called “double-hit signature” (DHITsig)—a signature originally described as being characteristic for high-grade B-cell lymphoma with MYC and BCL2 rearrangements (HGBCL-DH-BCL2). DHITsig was expressed in 21% of 431 germinal center B-cell-like (GCB)–DLBCL and all 55 Burkitt lymphomas examined. Reflecting this latter finding, DHITsig has been renamed the “dark zone signature” (DZsig). DZsigpos-DLBCL, non-DZsigpos GCB-DLBCL and activated B-cell-like (ABC)–DLBCL were associated with a 2 year overall survival of 57%, 89%, and 71%, respectively. 62% of DZsigpos tumors were negative for HGBCL-DH-BCL2 by FISH, but were associated with outcomes similar to HGBCL-DH-BCL2. A small group of HGBCL-DH-BCL2 that lacked DZsig expression had different molecular features compared with DZsig-expressing HGBCL-DH-BCL2 and were associated with favorable outcomes comparable to DLBCL, not otherwise specified. DZsigpos and ABC-DLBCL had a shorter diagnosis-to-treatment interval (DTI) than GCB-DLBCL, with this metric being associated with outcome. In conclusion, DZsig expression extends beyond HGBCL-DH-BCL2 and captures a poor-prognosis DLBCL subgroup with short DTI, including patients unidentifiable by routine FISH testing, that should be considered for treatment intensification or novel therapies in prospective trials.

EZH2 inhibitor DZNep blocks cell proliferation of GCB-DLBCL cells by upregulating p16

Diffuse large B-cell lymphomas (DLBCLs) are phenotypically and genetically heterogeneous. Two main subgroups of DLBCL include germinal center B-cell-like (GCB) and activated B-cell-like (ABC). Molecular profiling can further classify DLBCL into four subtypes: MCD (both CD79B and MYD88 L265P), BN2 (NOTCH2 mutation or BCL6 fusion), N1 (NOTCH1 mutation), or EZB (EZH2 mutation or BCL2 fusion). EZH2 inhibitors were recommended for patients with the EZB subtype of DLBCLs; however, little is known about the therapeutic mechanisms. Our results showed that DZNep arrested G1/S phase of GCB-DLBCL cells and inhibited the cell proliferation in vitro through upregulation of p16 by demethylating its promoter. These results suggest that DZNep may have potential as a novel therapeutic agent for DFLBL therapy. This agent may serve as a novel molecular agent to be applied to GCB DLBCL.

META-ANALYSIS DISPLAYED TRANSCRIPTOMIC SIGNATURE OF DIFFUSE LARGE B-CELL LYMPHOMA WITH SIGNIFICANT DIFFERENCES BETWEEN SUBTYPES

Lymphomas are cancers that affect the hematological system originated in the lymphatic system with an abnormal proliferation of lymphoid cells. Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma in adults. This lymphoma is well characterized by the presence of B-lineage lymphoid cells with immature features with diffuse proliferation in the affected tissue. This disease usually follows an aggressive clinical course, placing patients in immediate need of chemotherapeutic treatment. DLBCL can be further classified into two molecular subtypes, center-germinative B-cell (GCB) and activated B-cell (ABC) type lymphomas. This classification indicates the different stages of maturation of the neoplastic cells, and the different pathological mechanisms associated with them. The present work aimed to compare transcriptome signature of the center-germinative B-cell (GCB) and activated B-cell (ABC). Microarray studies were selected based on predefined criteria and downloaded from the GEO database. Data acquisition, processing, meta-data construction, as well as the differentially expressed genes were performed using R studio packages. Also, significantly enriched gene sets were identified using Gene Set Enrichment Analysis (GSEA). Gene expression and survival analysis from TCGA database were performed using the UALCAN and GEPIA webservers. From a total of 7,854 differentially expressed genes, 465 significantly down-regulated genes and 403 up-regulated genes in GCB subtype when compared to ABC subtype. Among these DEGs, in the GCB versus ABC comparison, the genes with the highest expression level were SPIN3 (FC: 1.12E+20), ZNF791 (FC: 9.43E+19) and TSPAN3 (FC: 9.18E+19), while the significant DEGs with the lowest expression level were KMT2C (FC: -3.09E+20), RIOX2 (FC: -1.49E+20) and ABHD11 (FC: -6.96E+19). Interestingly, only two genes had their expression levels significantly correlated to the DLBCL patient survival, the ABHD11 gene (p-value = 0.018) and MORF4L2 (p-value = 0.029). ABHD11 gene at low-medium level and MORF4L2 gene at high expression level significantly decrease the survival of patients. the meta-data generated from this study suggests that patients with diffuse large B-cell lymphoma pattern molecularly relevant gene expression in aspects related to cell proliferation, cell metabolism, and insensitivity to inhibitory signals to tumor progression as presented by the type of activated B cells. The findings converge the significant modulation of germinal center cells to gene expression regulation processes. Further experiments are still required, the transcriptional and translational activity, as well as cellular activity, appear to be decreased in the GCB type. Our findings, such as the one regarding the ABHD11 gene, which at low levels in GCB type, confers decreased survival rate in lymphoma patients. Therefore, this work brings important information for the understanding of the pathological process of the disease and its subtypes, and such biomarkers may serve as potential candidates for the development of molecular diagnostic methods and new therapeutic strategies for DLBCL subgroups.

The Alternative RelB NF-kB Subunit Exerts a Critical Survival Function upon Metabolic Stress in Diffuse Large B-Cell Lymphoma-Derived Cells.

Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma in adults and reveals distinct genetic and metabolic signatures. NF-κB transcription factor family is involved in diverse biological processes enabling tumor development and resistance to anticancer-therapy through activation of its two main pathways, the canonical and the alternative NF-κB pathways, the main actor of the latter being the RelB NF-kB subunit. RelB DNA binding activity is frequently activated in DLBCL patients and cell lines. RelB activation defines a new DLBCL subgroup with dismal outcome upon immunochemotherapy, and RelB confers DLBCL cell resistance to DNA damage. However, whether RelB can impact on DLBCL cell metabolism and survival upon metabolic stress is unknown. Here, we reveal that RelB controls DLBCL oxidative energetic metabolism. Accordingly, RelB inhibition reduce DLBCL mitochondrial ATP production, and sensitizes DLBCL cells to apoptosis induced by Metformin and L-asparaginase (®Kidrolase), two FDA approved antimetabolic drugs targeting mitochondrial metabolism. RelB also confers DLBCL cell resistance to glutamine deprivation, an essential amino acid that feeds the TCA cycle. Taken together, our findings uncover a new role for RelB in the regulation of DLBCL cell metabolism and DLBCL cell survival upon metabolic stress.

Differentiated approach in the treatment of diffuse large B-cell lymphoma according to c-MYC and BCL2 status

Background. The treatment results of patients with diffuse large B-cell lymphoma (DLBCL) with c-MYC proto-oncogene and BCL2 protein positivity according to the CHOP-21 ± R scheme remain unsatisfactory, which dictates the need to intensify the therapy regimen. Due to the fact that most authors combine DH (double hit) lymphomas and DE (double expressor) lymphomas for analysis, these data do not allow forming an accurate idea of the independent significance of the phenomenon of activated transcription factor c-MYC and BCL2 protein in patients with DLBCL and the results of treatment specifically in this group. Some researchers believe that the presence of c-MYC gene aberration and BCL2 protein expression are associated with negative clinical characteristics. Nevertheless, the clinical features of this subgroup of DLBCL remain poorly understood.The objective of the study was to analyze the incidence of c-MYC gene aberrations, BCL2 expression in DLBCL patients and to evaluate the effectiveness of intensive R + Hyper-CVAD chemotherapy regimen in this group of patients.Materials and methods. We analyzed the data of 80 patients aged 18–65 years with a newly diagnosed DLBCL for the period from 2018 to 2020. The diagnosis was established by histological and immunohistochemical examination. The tumor stages (II–IV) were assessed according to the Ann Arbor classification; the patient’s somatic status – according to the ECOG scale with a total of no more than 3 points. To detect c-MYC gene aberrations in the tumor tissue, fluorescent in situ hybridization was performed; to determine the BCL2 protein expression and Ki-67 proliferative index – an immunohistochemical study was performed. Patients with identified c-MYC gene aberrations received treatment, which included 8 courses of polychemotherapy according to the R + Hyper-CVAD scheme and high-dose chemotherapy according to the BFR scheme with autologous hematopoietic stem cell transplantation. Patients with DLBCL, in whom c-MYC gene aberration was not detected, were assigned to the group of intermediate and low malignancy. They received a polychemotherapy regimen according to the R-CHOP-21 scheme (up to 8 courses).Results and conclusion. For the first time in Kazakhstan, we studied the incidence of c-MYC proto-oncogene aberrations, BCL2 protein expression and proliferative activity (Ki-67) in DLBCL patients, which made it possible to identify groups of highly aggressive tumor and intermediate, low malignancy, apply personalized polychemotherapy programs and evaluate the immediate treatment efficacy. This approach made it possible to increase treatment effectiveness in the group of patients with highly aggressive DLBCL and to achieve comparable results with therapy in patients with a more favorable prognosis.

Role of local treatment in primary breast B-cell non-Hodgkin's lymphoma: a propensity score matching-based analysis from SEER database.

Primary breast lymphoma (PBL) has been defined as disease localized to breast with or without ipsilateral axillary nodal involvement. Primary breast B-cell non-Hodgkin's lymphoma is rare to be diagnosed clinically. The role of surgery and radiotherapy (RT) as local treatment is unclear. The aim of this study was to evaluate the prognostic factors and investigate the effect of local treatment in patients with primary breast B-cell non-Hodgkin's lymphoma. We identified patients with primary breast B-cell non-Hodgkin's lymphoma diagnosed between 1998 and 2015 in the Surveillance, Epidemiology, and End Results (SEER) database. Propensity score matching (PSM) was performed to reduce possible bias between groups. The overall survival (OS) and disease-specific survival (DSS) were calculated using the Kaplan-Meier method. Multivariate Cox regression analysis was used to identify independent prognostic factors. Altogether 956 patients with primary breast B-cell non-Hodgkin's lymphoma were included. Most patients were white women over the age of 60. The most common histological type was diffuse large B cell lymphoma (DLBCL), and most patients present with stage I disease. Furthermore, old age (>60 years), DLBCL histology and stage IIE disease were the statistically significant factors associated with worse OS and DSS. Surgery did not improve survival of patients, and surgery combined with RT did not achieve a better prognosis than RT alone. RT was associated with better survival in patients with stage IE DLBCL, but patients with stage IE MZL and FL and stage IIE primary breast B-cell non-Hodgkin's lymphoma could not benefit from RT. In local treatment, surgery offered no survival benefit for patients with primary breast B-cell non-Hodgkin's lymphoma, while RT is an effective choice because it can improve both OS and DSS in the stage IE DLBCL subgroup.