Survival In Diffuse Large B-cell Lymphoma Research Articles

The prognostic significance of MYC/BCL2 double expression in DLBCL in the genetic classification era.

Double expression (DE) is a World Health Organization-recognized adverse prognostic factor in diffuse large B-cell lymphoma (DLBCL). However, the prognostic value of DE in the genetic subtyping era and potential mechanisms remain to be explored. We enrolled 246 DLBCL patients diagnosed between December 2021 and September 2023 in a Jiangsu Province Hospital cohort and included 930 DLBCL patients from three published studies in an external cohort. Double-expression DLBCL (DEL) in the external cohort was mainly distributed in the OTHER subtype (42.0%), EZB subtype (28.3%), and MCD subtype (15.0%), whereas the MCD subtype exhibited the highest ratio of DEL. DEL was significantly related to unfavorable overall survival (OS) and progression-free survival (PFS) in DLBCL, but only in EZB and OTHER subtypes that DEL retained remarkably adverse impacts on survivals compared to non-DEL. We explored the prognostic value of clinical and genetic parameters in DEL patients and found only ST2 showed better OS than A53 in DEL patients, whereas the other subtypes showed no significant difference. DEL showed similarities with the MCD subtype in mutation profiles. Furthermore, RNA-sequencing analyses exhibited upregulation in tumor proliferation-related pathways in DEL patients, but downregulation in extracellular matrix organization, T-cell activation and proliferation, type II interferon production, and pathways associated with cell death might contribute to the poor outcomes of DEL patients.

DA-R-EPOCH May Mitigate the Adverse Prognostic Implication of the Diagnosis-to-Treatment Interval (DTI) in Large B-Cell Lymphomas

BackgroundShort diagnosis-to-treatment interval (DTI) is associated with high-risk disease and poor survival in diffuse large B-cell lymphoma (DLBCL). There is a paucity of literature on DTI and survival in DLBCL treated with first-line DA-R-EPOCH. We hypothesized that rapid initiation of DA-R-EPOCH in aggressive and high-risk DLBCL mitigates the adverse prognostic implication of short DTI. Patients and MethodsWe retrospectively examined the association of DTI, categorically (short DTI ≤ 14 and long > 14 days) and continuously, with clinical features and survival outcomes in DLBCL treated with first-line DA-R-EPOCH at our institution. ResultsA total 190 patients were analyzed, 21% with high-grade DLBCL subtypes, 56% IPI ≥ 3, and median DTI of 13 days. The short DTI cohort contained more patients with IPI ≥ 3, bulky disease, and elevated LDH. When analyzed categorically and continuously, DTI was not associated with significant differences in PFS or OS. There was significant multivariable interaction between bulky disease, DTI, and PFS (P = .033), with improved PFS in patients with bulky disease in the short DTI cohort. ConclusionWe found that negative prognostic implications of DTI are mitigated in DLBCL patients treated with first-line DA-R-EPOCH, suggesting that urgent initiation of DA-R-EPOCH in high-risk DLBCL, including bulky disease, may improve survival. Our study's shorter DTI compared with DTIs reported in prospective DLBCL trials highlights DTI as a marker of external validity in clinical trial results. Future trials should implement protocols encouraging shorter, realistic DTIs to avoid selection bias against high-risk patients who are unable to delay treatment.

An interpretable survival model for diffuse large B-cell lymphoma patients using a biologically informed visible neural network

Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma (NHL) and is characterized by high heterogeneity. Assessment of its prognosis and genetic subtyping hold significant clinical implications. However, existing DLBCL prognostic models are mainly based on transcriptomic profiles, while genetic variation detection is more commonly used in clinical practice. In addition, current clustering-based subtyping methods mostly focus on genes with high mutation frequencies, providing insufficient explanations for the heterogeneity of DLBCL. Here, we proposed VNNSurv (https://bio-web1.nscc-gz.cn/app/VNNSurv), a survival model for DLBCL patients based on a biologically informed visible neural network (VNN). VNNSurv achieved an average C-index of 0.72 on the cross-validation set (HMRN cohort, n = 928), outperforming the baseline methods. The remarkable interpretability of VNNSurv facilitated the identification of the most impactful genes and the underlying pathways through which they act on patient outcomes. When only the 30 highest-impact genes were used as genetic input, the overall performance of VNNSurv improved, and a C-index of 0.70 was achieved on the external TCGA cohort (n = 48). Leveraging these high-impact genes, including 16 genes with low (<5 %) alteration frequencies, we devised a genetic-based prognostic index (GPI) for risk stratification and a subtype identification method. We stratified the patient group according to the International Prognostic Index (IPI) into three risk grades with significant prognostic differences. Furthermore, the defined subtypes exhibited greater prognostic consistency than clustering-based methods. Broadly, VNNSurv is a valuable DLBCL survival model. Its high interpretability has significant value for precision medicine, and its framework is scalable to other diseases.

Abstract PO-034: Targeting metabolic plasticity and flexibility via SIRT3 inhibition to restrain nucleotides availability in DLBCLs

Abstract Cancer cells need metabolic adaptations to survive in hostile environment such as low nutrient and drug treatment conditions. DLBCLs (Diffuse large B cell lymphoma) originate from a metabolic competitive/challenging environment of germinal center (GC) when B cells undergo positive selections against antigens. However, it is largely unknown whether and how metabolic flexibility (the ability to use different nutrients) and plasticity (the ability to process the same nutrient differently) contribute to DLBCLs. Our previous study identified that SIRT3, the only mitochondrial protein deacetylase, is critical to survival and proliferation of DLBCLs by promoting glutaminolysis. Our mitochondrial localized SIRT3 inhibitor, YC8-02, has pan-toxicity to all DLBCL cells tested regardless of different subtypes with somatic mutations. Here, we observed glucose as an alternative nutrient source to compensate in the TCA cycle in SIRT3 deficient cells. DLBCLs exhibited resistance to SIRT3 inhibition in high glucose condition, while SIRT3 inhibited DLBCLs highly depend on glucose metabolism. The combination of YC8-02 with glucose metabolism inhibitors induces synergistic killing to DLBCLs. As glucose is the main carbon source of cells, it is unknown how DLBCLs manage the flexibility of using glucose for different purposes. Using in vivo metabolic imaging (MALDI), we observed that, in contrast to normal GC B cells, lymphomas specifically use the glucose for de novo nucleotide synthesis. SIRT3 inhibition decreased nucleotide levels in DLBCL cells in vitro and in vivo. The metabolic flux tracing showed that DLBCL cells requires SIRT3 activity to activate a glucose/serine/one-carbon metabolism pathway for de novo nucleotide biosynthesis, and Inhibition of SIRT3 decreased the glucose and serine being used for nucleotides in DLBCLs. Serine as another potent source for nucleotides can rescue DLBCLs from SIRT3 inhibition. Furthermore, we validated that overexpression of enzymes, like MTHFD2, in one carbon metabolism pathway can rescue the proliferation in SIRT3 deficient cells. High nucleotides levels in medium also help DLBCLs survival in YC8-02 treatments. To overcome the metabolic flexibility on environmental nucleotides, we used inhibitors of nucleoside transporters (ENT1/2), dipyridamole and draflazine, which can synergize with YC8-02 to kill DLBCLs in vitro and eradicate systemic lymphomas in vivo. In summary, our results illustrate that DLBCLs with active SIRT3 exhibit metabolic plasticity by repurposing glucose to nucleotide biosynthesis, while lymphoma cells can also reply on metabolic flexibility to manage nutrient usage and gain extracellular nucleotides for survival under SIRT3 inhibition. Our combination treatment SIRT3 and ENT inhibitors can be a promising therapy for DLBCLs by restraining their metabolic plasticity and flexibility. Citation Format: Meng Li, Noel R Park, Chris Chin, Coraline Mlynarczyk, Hening Lin, Shawn Davidson, Ari M Melnick. Targeting metabolic plasticity and flexibility via SIRT3 inhibition to restrain nucleotides availability in DLBCLs [abstract]. In: Proceedings of the Fourth AACR International Meeting on Advances in Malignant Lymphoma: Maximizing the Basic-Translational Interface for Clinical Application; 2024 Jun 19-22; Philadelphia, PA. Philadelphia (PA): AACR; Blood Cancer Discov 2024;5(3_Suppl):Abstract nr PO-034.

Abstract 4839: Ethnicity dictates survival benefit in Hispanics diagnosed with diffuse large B-Cell lymphoma survival: A national cancer database analysis

Abstract Background: Diffuse Large B-Cell Lymphoma (DLBCL) is the most common Non-Hodgkin lymphoma in the United States (US) but there is limited evidence showing ethnic disparities affecting overall survival (OS) (Blood10.1182/blood-2022-159201, Blood10.1182/blood-2021-151816). This is the largest nationwide cancer registry analysis evaluating ethnic differences for HI vs Non-Hispanics (NH) with DLBCL in the US. Methods: Data were analyzed using the National Cancer Database from 2004-2019. Sociodemographic characteristics were compared between ethnic groups. Kaplan-Meier and Cox regression analyses were used to compare OS between HI and NH. Multivariate analysis and propensity score matching were performed with adjustment for age, stage, comorbidity score, and insurance status, type of facility, and great circle distance Results: 239,391 patients (HI n=18,290, NH n=221,101) were diagnosed with DLBCL. Male sex predominated for both and the majority of patients were Whites. HI were diagnosed at a median age of 62 years (y) vs 68 y for NH [p&amp;lt;0.001]. For HI and NH, most of the patients were diagnosed from 2016-2019. For both groups, the majority of patients had a Charlson-Deyo Score of 0, stage IV at diagnosis and unknown HIV status. The primary payer at diagnosis was government sponsored for HI and NH. In HI the median income based on the Median Income Quartile for 2000 corresponded to the highest level of &amp;gt;$46,000 and from 2008-2012 it was similarly distributed. For NH, for both time periods the majority of the patient were in the highest bracket. Most patients in the HI and NH were located in metropolitan area and were more likely to be treated at a comprehensive cancer center with a great circle distance (miles) of 7.3 for HI and 9.7 for NH. The median survival for HI was 11 y vs 6.8 y for NH. The survival probability at 2, 5 and 10 y for HI corresponded to 69%, 61% and 52%, while for NH it was 66%, 55% and 41%, respectively. The OS probability at 10 y was statistically significant favoring HI [p&amp;lt;0.0001]. Independently, on multivariate analysis, not insured status was associated with worse OS (HR 1.2, CI 1.13-1.28, [p&amp;lt;0.01]) and private insurance type was associated with better OS (HR 0.81, CI 0.77-0.84, [p&amp;lt;0.01]) Conclusion: This nationwide cancer registry study identified better OS in HI diagnosed with DLBCL in the US. Both cohorts had similar sociodemographic and clinical results; however, HI were diagnosed at younger age. This finding may not only help to explain a unique trait towards development of cancer at an earlier age, but also an enhanced response to therapy. Regarding median income there wasn’t a tendency noted for HI, which support the unique complex interactions in socioeconomic status. Further studies examining intrinsic biologic differences are needed to better understand improved OS for HI, which could also help the rational development of targeted therapies. Citation Format: Carolina Velez-Mejia, Esteban Toro Velez, Daniel Rosas, Qianqian Liu, Joel E. Michalek, Enrique Diaz Duque. Ethnicity dictates survival benefit in Hispanics diagnosed with diffuse large B-Cell lymphoma survival: A national cancer database analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4839.

Abstract 3648: The spatial organization of cells expressing MYC and BCL2 affects immune microenvironment composition and prognosis in DLBCL

Abstract Point process analyses are widely used in ecology and geography to understand the impact and relevance of spatial distribution of points in a complex system. To date, only few studies have explored point process analyses in the context of tumor heterogeneity. Malignancies often show variable patterns of cellular distribution, and the relationship of these topographic variables with underlying biological processes and clinical outcomes is not well understood. Here, we performed spatial point process analyses in diffuse large B-cell lymphoma (DLBCL), leveraging advances in multiplexed immunohistochemistry and cellular phenotyping, with downstream multi-omic and clinicopathological analyses. We have demonstrated that survival in DLBCL is strongly associated with the fraction of malignant cells showing co-expression of the oncogenes MYC and BCL2 in the absence of BCL6 (M+2+6-). These cells display non-random spatial organization within tumor infiltrates, deviating from a poisson distribution. We therefore modeled the spatial organization of M+2+6- cells within DLBCL, using x-y coordinate information from multiplexed fluorescent immunohistochemistry (mfIHC) in four independent cohorts (N=449 patients). We derived spatial point patterns, upon which Geyers point process models were applied. We observed that patients could be divided into two groups based on these: one group showed “clustered” spatial organization, while the other displayed a “dispersed” M+2+6- cell distribution. Cases with predominantly “dispersed” M+2+6- cells showed shorter overall survival in all analyzed cohorts (P &amp;lt; 0.05 in 4/4 cohorts). We performed multi-omic analyses to identify potential biological explanations for this association between topography and clinical outcome. Using digital spatial profiling of the transcriptome from CD20, CD3 and CD68 compartments in 47 DLBCL samples, we observed that lymphomas with an M+2+6- ‘dispersed’ phenotype had an immunologically cold microenvironment enriched in Tregs and exhausted CD4+ and CD8+ T cells. Through an integration with single-cell transcriptomic analyses (N=22 samples) we observed that malignant B-cells from cases with M+2+6- ‘dispersed’ phenotype expressed genes implicated in cell migration and adhesion, but also in potentially targetable immune checkpoints such as LAG3. In summary, by integrating multiplexed immunophenotyping and point process analysis with molecular profiling in the setting of DLBCL, we show that a dispersed pattern of distribution of the M+2+6- subclonal fraction uniquely confers poor prognosis, and associates with a potentially reversible immune cold microenvironment composition. This represents the first demonstration that the spatial distribution of malignant cell subpopulations in lymphoma can embody biological and clinical significance. Citation Format: Shruti Shridar, Michal Marek Hoppe, Patrick Jaynes, Gayatri Kumar, Siddham Jasoria, Ziwei Meng, Vaibhav Rajan, Kasthuri Kannan, Claudio Tripodo, Anand Devaprasath Jeyasekharan. The spatial organization of cells expressing MYC and BCL2 affects immune microenvironment composition and prognosis in DLBCL [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3648.

Are changes in patient-reported outcomes prognostic for diffuse large B-Cell lymphoma survival? Results from the GOYA trial.

In this hypothesis-generating analysis, we examined whether longitudinal changes in patient-reported outcomes (PROs), such as symptoms, over time would be prognostic for progression-free survival (PFS) and overall survival (OS) in patients with diffuse large B-cell lymphoma (DLBCL) who were newly treated with obinutuzumab (G) in combination with CHOP (G-CHOP) or rituximab (R) with CHOP (R-CHOP), in the GOYA Phase 3 trial (NCT01287741). Our results show that from the study baseline to cycle 3day 1, every 1-point increase (worsening) in fever symptoms was associated with a 41% higher risk of death (hazard ratio [HR], 1.41; P = 0.01). Every 1-point increase (worsening) in lumps or swelling symptoms was associated with a 27% higher risk of disease progression or death (PFS events) (HR, 1.27; P = 0.01) and a 29% higher risk of death (OS events) (HR, 1.29; P = 0.02). No significant associations were observed between survival and changes in other symptoms, such as itching. Our study suggests that changes in some PROs are related to survival in DLBCL patients.

Spatially-resolved transcriptomics reveal macrophage heterogeneity and prognostic significance in diffuse large B-cell lymphoma

Macrophages are abundant immune cells in the microenvironment of diffuse large B-cell lymphoma (DLBCL). Macrophage estimation by immunohistochemistry shows varying prognostic significance across studies in DLBCL, and does not provide a comprehensive analysis of macrophage subtypes. Here, using digital spatial profiling with whole transcriptome analysis of CD68+ cells, we characterize macrophages in distinct spatial niches of reactive lymphoid tissues (RLTs) and DLBCL. We reveal transcriptomic differences between macrophages within RLTs (light zone /dark zone, germinal center/ interfollicular), and between disease states (RLTs/ DLBCL), which we then use to generate six spatially-derived macrophage signatures (MacroSigs). We proceed to interrogate these MacroSigs in macrophage and DLBCL single-cell RNA-sequencing datasets, and in gene-expression data from multiple DLBCL cohorts. We show that specific MacroSigs are associated with cell-of-origin subtypes and overall survival in DLBCL. This study provides a spatially-resolved whole-transcriptome atlas of macrophages in reactive and malignant lymphoid tissues, showing biological and clinical significance.

Non-neoplastic cells as prognostic biomarkers in diffuse large B-cell lymphoma: A system review and meta-analysis.

The microenvironment of diffuse large B-cell lymphoma (DLBCL) is composed of various components, including immune cells and immune checkpoints, some of which have been correlated with the prognosis of DLBCL, but their results remain controversial. Therefore, we conducted a systematic review and meta-analysis to investigate the association between the microenvironment and prognosis in DLBCL. We searched PubMed, Web of Science, and EMBASE for relevant articles between 2001 and 2022. Twenty-five studies involving 4495 patients with DLBCL were included in the analysis. This meta-analysis confirmed that high densities of Foxp3+Tregs and PD-1+T cells are good indicators for overall survival (OS) in DLBCL, while high densities of programmed cell death protein ligand1(PD-L1)-positive expression cells and T-cell immunoglobulin-and mucin domain-3-containing molecule 3 (TIM-3)-positive expression tumor-infiltrating cells (TILs) play a contrary role in OS. Additionally, higher numbers of T-cell intracytoplasmic antigen-1(TIA-1)-positive expression T cells imply better OS and progression-free survival (PFS), while high numbers of lymphocyte activation gene(LAG)-positive expression TILs predict bad OS and PFS. Various non-tumoral cells in the microenvironment play important roles in the prognosis of DLBCL.

Diffuse Large B Cell Lymphoma: Immunohistochemical Classification According to Hans Algorithm and Association With Outcome in A Moroccan Institution.

The most prevalent subtype of non-Hodgkin lymphoma is diffuse large B-cell lymphoma (DLBCL). Germinal center B-cell (GCB) and non-germinal center B-cell (non GCB) are the two main biologically different molecular subtypes identified utilizing an immunohistochemistry-based approach. Our objective in this study is to analyze the impact of immunohistochemical subtypes of DLBCL (GCB or non GCB) on demographic and clinicopathological parameters, response to chemotherapy and survival outcomes. This is a retrospective study including 106 cases of DLBCL collected in the department of pathology, Hassan II university hospital, Fez (Morocco), over a period of 12 years (January 2010-September 2022). The subtypes of DLBCLs were defined according to Hans algorithm, using immunohistochemistry by three biomarkers (CD10, BCL6, MUM1). Independent t tests and analyses of variance were used for the comparison of mean values. We employed the SPSS 26.0 program to achieve this. A statistically significant value was set at P < .05. Seventy-five patients (71%) were non-GCB subtype, while thirty-one patients (29%) had the GCB immunosubtype. We have found a significant (P < .05) correlations between DLBCL immunosubtypes and treatment responses on one hand and survival in the other hand. In the GCB subtype, the response rate and survival were significantly improved. A significant association was found between Ki 67 expression and survival on univariate analysis. On multivariate analysis, we note a correlation between Ki 67 expression, DLBCL immunohistochemical subtypes and survival outcome. Non GCB subtype is associated with poor response to treatment and inferior survival outcome compared to GCB subtype in Moroccan context, especially when combined with high expression of Ki 67 marker.

Abstract C053: Multi-omics guided design of lymphoid organoids to study cooperative cell signaling in lymphomas

Abstract Our collaborative research (Cell1) used 1001 patients of diffuse large B-cell lymphoma (DLBCL) to show that genetic alterations interact with clinical factors to impact overall survival (OS) in DLBCL. B cell receptor (BCR) signaling-driven activated B cell-like (ABC) DLBCL subtypes is the most chemo-resistant subtype to the frontline therapy R-CHOP, with 40% of patients experiencing no response or relapse. We have systematically examined disparities in lymphoma outcomes and identified African-American (AA) patients as a poor-risk population in DLBCL, with a diagnosis age 10 years younger than other racial groups. We found that AA-DLBCL patients experienced inferior 5-year OS compared to non-Hispanic whites (38% vs 46%)2,3. We performed the first-ever characterization of genetic alterations among AA-DLBCL patients and demonstrated distinct mutation patterns across DLBCL arising in AA ancestry groups4. While some of the most frequent genetic mutations in AA-DLBCL are related to chromatin and epigenetics, few studies have investigated racial disparities in patients with aggressive ABC DLBCLs. The mechanisms that determine this racial inequality are unknown but may lie in the particular spectrum of mutations that act in concert with intricate survival signals imparted by the lymph node tumor microenvironment (Ly-TME). The pathogenesis of ABC DLBCLs has been linked to constitutive activation of and cooperation between the BCR and Toll-like receptor (TLR) pathways. These pathways are emerging as novel therapeutic targets, with a particular interest in MALT1 (mucosa-associated lymphoid tissue translocation protein 1). However, the genetic heterogeneity of ABC DLBCLs and the cooperative signaling mediated by the Ly-TME have led to discrepancies in drug responses. As a first step towards understanding Ly-TME in DLBCLs (Nature Materials5), we used DLBCL patient samples from multiple cohorts, covering &amp;gt;1100 samples and imaging analysis to determine immune and ECM cues in the lymphoid TME. These samples were not classified into racial categories. We designed a biomaterials-based modular lymphoma organoid that presents specific integrin ligands at controlled densities to DLBCLs. Human ABC-DLBCLs responded to MALT1 inhibition in an integrin-ligand-dependent manner. TLR activation with TLR agonists rescued cells from MALT1 inhibitors, possibly due to increased MALT1 expression. The inclusion of T-cell signals increased MALT1 expression and rescued cells from MALT1 inhibitor-mediated killing in cell lines and human PDX organoids. Changes in RNA-seq and phosphorylation in the TLR &amp; BCR pathways were used to inform effective combination therapy approaches with MALT1 inhibitors in vitro and in vivo5. Ongoing work is determining the racial disparities in AA lymphomas through complex interactions between Ly-TME and DLBCLs.

Machine Learning Can Outperform Ann Arbor Staging in Predicting Survival in Patient with Diffuse Large B-Cell Lymphoma: Analysis of a Large National Cancer Database

Introduction: Diffuse Large B-cell Lymphoma (DLBCL) is the most common lymphoma in the world with usually an aggressive clinical course. The Ann Arbor staging system and International Prognostic Index (IPI) commonly utilized in clinical practice for risk stratification have known limitations. Machine learning (ML) has emerged as a promising tool for more comprehensive and deeper data analysis. We sought to utilize the ability of ML to predict survival in DLBCL compared to Ann Arbor staging system using a large national database. Methodology: We employed the ML algorithm XGBoost on the National Cancer Institute's Surveillance, Epidemiology and End Result (SEER) database to predict overall survival (OS) and the lymphoma specific survival (LSS). For prediction analysis, we transformed the survival labels into a simple Boolean format: “alive” represented as 0, “dead” as 1, and “dead (attributable to this cancer diagnosis)” also as 1. We utilized one-hot encoding to convert categorical features and variables into binary vectors. The data set was divided into two parts: training (80%) and test (20%). Further, we split the training set into the actual training set and validation set by using stratified 5-fold cross validation. Hyper-parameter optimization was done within the validation set. A broad range of attributes were utilized by the model for its prediction algorithm. To understand how each attribute contributes to predictions, we calculated its importance score in XGBoost. Results: A total of 64,912 patients with DLBCL were found and their data were extracted. The majority were Caucasian (78.9%) with a median age range of 60 to 69. The model was able to predict OS and LSS, with an area under the curve (AUC) of 0.89 and 0.75 (Figure 1), respectively. Factors selected by the model for survival prediction included presence or absence of B-symptoms, treatment status, and disease stage. For OS and LSS, the model found B symptoms to be the highest contributing factor with an importance score of 0.205 and 0.167, respectively. Other important factors incorporated by the model included age and stage IV for OS, and stage IV and clinically asymptomatic status for LSS. The least important factors were location of the primary lymphoma site and year of diagnosis (Table 1). Conclusion: Machine learning tools can help predict survival in patients with DLBCL and able to challenge current staging systems. Our results warrant validation in future prospective studies.

Validating the Halp Score (Hemoglobin,Albumin, Lymphocytes, and Platelets) and the Neutrophil/Lymphocyte Ratio ( NLR) As Prognostic Factors for Overall Survival in Patients with Diffuse Large B-Cell Lymphoma. Retrospective Analysis By the Grupo De Estudio De Latino America De Linfoproliferativos (GELL)

Introduction. Diffuse large B-cell lymphoma ( DLBCL) is the most common type of malignant lymphoid neoplasm. The international Prognostic Index (IPI) and its variants are the main prognostic tools used in DLBCL with value in the rituximab era. Certain molecular biomarkers and genetic signatures in DLBCL have been identified, but the cost and unavailability in Latin America ( LATAM) are an issue. Therefore, using accesible tools in LATAM is a potencially unmet need. Vlatka (2021), reported a new score (HALP: hemoglobin, albumin, lymphocytes, and platelets) for overall survival (OS) in DLBCL. These parameters are easy to access in LATAM centers. Thus, we aim to validate this novel OS score in the GELL database, and expanding database from LATAM countries. Methods. This was a retrospective analyses of patients with de novo DLBCLtreated with curative intent between 2010 and 2018. The classic 5 international prognostic index variables [age, ECOG, Extranodal (EN) involvement, Lactate dehydrogenase (LDH), and advanced stage] and low serum albumin defined as ≤3.5mg/dL(divided in low, 3.4-2.5 mg/dL, and very low, ≤2.4 mg/dL), as previously described by our group ( Villela,2018&amp;2019). We also calculated the NLR considering an adverse prognostic factor &amp;gt;4, as previously published by us ( Beltran&amp;Villela,2020). The HALP score was calculated using the following formula: hemoglobin(g/L) x albumin (g/L) x absolute lymphocyte count (k/μL) divided by platelets (k/μL). The ROC method was used to calculate the HALP cut-off. Demographic characteristics are reported using descriptive statistics. Cox proportional-hazard regression model was used to evaluate parameters associated with OS, and survival curves were estimated with the Kaplan-Meier (KM) method. Outcomes. 1407 patients were included, who were treated with standard RCHOP (n=1112,79%), RminiCHOP (n=93,7%), REPOCH (n=111,8%), and CHOP (n=91,6%). The median follow-up was 36 months (IQR: 7 to 56). NLR &amp;gt;4 was observed in 18.5%. The median HALP score was 24 (IQR, 12 to 40), and the cut-off of ≤13 (AUC 0.58;95%CI 0.56 to 0.61; p&amp;lt;0.0001) was considered an adverse prognostic factor, which was observed in 451 patients (32.7%). Female sex, ECOG &amp;gt;1, EN &amp;gt;1. high LDH, advance stage, low and very low albumin, and NLR&amp;gt;4 were associated with HALP≤13, but age was not ( Table 1). Patients with HALP ≤13 had a lower 3-years OS rate than HALP&amp;gt;13 (48% vs. 66%, respectively; p&amp;lt;0.001). Table 2 shows the univariate &amp; multivariate analysis of the variables with independent influence on OS, including HALP &amp; NLR. In multivariate analysis, HALP and EN involvement were left out of the model ( Harrell´s C-Index, 0.73;95%CI 0.69 to 0.76). Conclusion: NLR&amp;gt;4, but not HALP ≤13, could prognosticate inferior OS in LATAM patients with DLBCL treated with curative intent. The adverse prognostic value of NLR&amp;gt;4 should be validated prospectively in other cohort of DLBCL patients.

Diagnosis to Treatment Interval Is the Most Significant Prognostic Factor Compared to Symptoms to Treatment As Well As to Contact to Treatment Intervals in DLBCL Patients

Background Short interval from diagnosis (i.e., biopsy) to treatment initiation (DTI) is associated with aggressive behavior of lymphomas including diffuse large B-cell lymphoma (DLBCL), and it should be considered as an inclusion criterium in clinical trials to avoid bias given by treatment delay. However, date of biopsy (diagnosis date) could be influenced by other factors. Initial symptoms to treatment interval (STI) as well as interval between the first contact with health care system and treatmet (CTI) could have impact on survival as well as and there is no data analyzing the impact of those intervals. The aim of this analysis was to understand the correlations among these intervals and the impact on survival of patients (pts) in the real-life setting. Methods Altogether 1576 pts with de novo DLBCL or high-grade B-cell lymphoma from Charles University, General Hospital, Prague, Czech Republic, were identified as part of the lymphoma project (NiHiL; NCT03199066). Of them 929 pts were diagnosed between 2010-2021, and 602 (65%) pts who were treated by R-CHOP in this period entered this retrospective analysis. STI ( n = 586), CTI ( n = 535), DTI ( n = 602) were analyzed together with baseline characteristics and survival of the pts (OS, EFS). Results The 602 pts (median age 64 years) were diagnosed with advanced clinical stage (aCS) in 64%, extranodal (EN) involvement ≥ 2 sites in 40%, PS ECOG ≥ 2 in 26%, elevated LDH in 61%, bulky disease ≥ 7.5 cm in 41%, B-symptoms in 42%, and IPI 3-5 in 52%. Median follow-up was 7.5 years (range 0.1-13.2 years). Median STI was 15 weeks (w) (interquartile range, IQR 10-24 w), CTI 9 w (IQR 6-14 w), DTI 32 days - 5 w (IQR 3-7 w, Figure). Significant correlations among the intervals ( P &amp;lt;0.001 for all correlations; coefficient 0.796 for STI x CTI, 0.307 for STI x DTI, and 0.372 for CTI x DTI) were observed. Shorter intervals to therapy, most significantly DTI, correlated with aggressive features of DLBCL, i.e., PS ECOG ≥ 2 (CTI P= 0.002, DTI P &amp;lt;0.001), elevated LDH (CTI P= 0.022, DTI P &amp;lt;0.001), B-symptoms (CTI P= 0.034, DTI P &amp;lt;0.001), aCS (DTI P &amp;lt;0.001), extranodal involvement (DTI P &amp;lt;0.001), IPI 3-5 (DTI P &amp;lt;0.001), and bulky disease (STI P= 0.005, CTI P= 0.011, DTI P &amp;lt;0.001). Out of 307 pts with short DTI (≤ 32 days) we identified 183 (60%) pts with STI &amp;gt; 15 w; these pts didn't have different outcome in comparison to those with shorter both STI and DTI. A pre-phase treatment ( n = 144) was associated with shorter intervals to therapy (STI P= 0.025, CTI P &amp;lt;0.001, DTI P &amp;lt;0.001) and inferior survival (EFS24 58% vs. 81%, HR 2.03, P &amp;lt;0.001; OS24 71% vs. 87%, HR 2.04, P &amp;lt;0.001). Shorter DTI was observed in pts who didn't respond to therapy in comparison to those who responded (med. DTI 27 vs. 33 days; P= 0.038). Shorter intervals to therapy were associated with significantly inferior survival, i.e., for STI 0-7 w vs. &amp;gt; 7 w (EFS24 66% vs. 77%, HR 1.53, P= 0.018; OS24 75% vs. 85%, HR 1.50, P= 0.087; with no significant survival difference between groups with STI 0-15 w vs &amp;gt; 15 w), for CTI 0-9 w vs. &amp;gt; 9 w (EFS24 69% vs. 82%, HR 1.53, P= 0.003; OS24 78% vs. 90%, HR 1.64, P= 0.002), and for DTI 0-4 w vs. &amp;gt; 4 w (EFS24 69% vs. 82%, HR 1.53, P&amp;lt; 0.001; OS24 78% vs. 90%, HR 1.67, P= 0.007). Multivariate analysis revealed all 3 intervals to be independent risk factors of survival when analyzed separately with IPI and gender (i.e., for EFS STI P= 0.039, CTI P= 0.029, DTI P= 0.028). DTI was however the only independent factor ( P= 0.001) among the 3 intervals when they were analyzed together. Conclusion Short intervals to therapy are independently associated with aggressive features of DLBCL and inferior survival. Despite multiple external factors that might influence the date of biopsy, DTI seems to be the most accurate predictor of survival and response to therapy. In pts with short DTI and long STI, no survival difference was observed when compared to other short-DTI pts indicating that these pts should be considered as high-risk similar to other short-DTI pts. This work was supported by the Charles University Hematology-Oncology Cooperatio Program and grant NU21-03-00411.

Clinical Outcomes of Patients with Newly Diagnosed Diffuse Large B-Cell Lymphoma in a County Hospital System

Introduction: Advances in the treatment of diffuse large B-cell lymphoma (DLBCL) have improved survival, with Surveillance, Epidemiology, and End Results (SEER) Program data showing 5-year overall survival (OS) of 64.7% (SEER*Explorer, 2023). However, insurance status in the US is related to DLBCL survival (Han et al., Cancer 2014), and real-world data for patients who are uninsured or who may have socioeconomic barriers to care remain limited. We performed a retrospective cohort study to examine outcomes for patients with newly diagnosed DLBCL treated in a large safety-net hospital system in the third most populous county in the US. Methods: Patients aged 18 years and older diagnosed with DLBCL between January 2011 and June 2022 and treated at Harris Health System, Houston, TX, were included in the analysis. Demographic, disease, treatment, response, and follow-up data were abstracted from the electronic medical record. US Census Block Group FIPS codes derived from geocoded home address at the time of initial diagnosis were used to compute the Area Deprivation Index (ADI). Eligibility for cellular therapies including hematopoietic stem cell transplantation (HSCT) and chimeric antigen receptor T-cell therapy (CAR-T) was determined by the abstractors. Time-to-event analysis was performed using the Kaplan-Meier method, and Cox proportional hazards regression was used to examine factors associated with event-free survival (EFS) and OS. Results: Among 461 patients with newly diagnosed DLBCL (Table 1), the median age was 53 years (range, 18-93), 75% were uninsured, and 80% were in the most disadvantaged ADI national quartiles, defined as at or above the fiftieth percentile. Most patients had stage III or IV disease and an Eastern Cooperative Oncology Group Performance Status (ECOG) of 0 to 1 at the time of diagnosis. One hundred ninety-eight patients (44%) had poor-risk disease by the Revised International Prognostic Index (R-IPI). Seventy-five (16%) had a history of HIV infection. Common first-line treatments included R-CHOP ( n = 210, 46%) and R-EPOCH ( n = 183, 40%); 18 received no therapy (3.9%). The median time from diagnosis to treatment initiation (DTI) was 18 days (range, 0-246); 76% completed therapy. Of 425 evaluable patients, 288 (68%) achieved a complete response (CR) and 44 (10%) had a partial response (PR); of these, 74 (22%) relapsed. Twenty-five (58%) of 43 patients eligible for cellular therapies at relapse received them: 14 (56%) underwent autologous HSCT, 8 (32%) received CAR-T, and 3 (12%) had both. Median follow-up time for the cohort was 48.8 months (range, 0.2-147). By the time of analysis, 167 patients (36%) had developed refractory/relapsed disease, and 148 (32%) had died. At 1, 2, and 5 years, EFS was 72%, 64%, and 58%, and OS was 82%, 76%, and 69% (Figure 1). At 1, 2 and 5 years, EFS was 77%, 69%, and 64% and OS was 85%, 79%, and 75% for patients with germinal center B-cell-like (GCB) DLBCL, whereas EFS was 68%, 56%, and 50% and OS was 80%, 73%, and 62% for those with non-GCB DLBCL. In multivariable Cox regression models, predictors of OS were poor-risk disease by R-IPI (HR 6.08 compared to very-good-risk disease, 95% CI 1.02-36.09), ECOG (HR 2.10 for ECOG ≥2 compared to 0-1, 95% CI 1.12-3.95), and normalized LDH (HR 1.07 per fold increase, 95% CI 1.03-1.11). Race/ethnicity, insurance status, and DTI were not associated with significant differences in OS. However, receiving fewer than planned cycles of first-line therapy was associated with higher mortality (HR 2.00, 95% CI 1.12-3.56), as was not attaining CR by the end of first-line therapy (HR 7.90, 95% CI 3.65-17.12). Conclusion: This is one of the largest real-world studies to date examining outcomes of patients with newly diagnosed DLBCL at a safety-net hospital system. Overall, patients had shorter EFS and OS compared to historical clinical trials but had similar OS to SEER estimates. In this health system with robust financial assistance programs and access to cellular therapies for some patients, uninsured status was not associated with inferior outcomes. Addressing barriers to care may improve outcomes for uninsured patients in other settings.

Baseline 18f-Fluorodeoxyglucose PET/CT Radiomics Predicts Outcome in Newly Diagnosed Intravascular Large B-Cell Lymphoma

[Introduction] Treatment outcomes of intravascular large B-cell lymphoma (IVLBCL), a rare subtype of extra-nodal diffuse large B-cell lymphoma (DLBCL), have dramatically improved in recent years. However, prognostic factors for IVLBCL are largely unknown. Several studies have explored the potential of the baseline 18F-fluorodeoxyglucose (FDG) PET/CT radiomics features to identify patients with poor prognosis. For example, metabolic tumor volume (MTV) reflects the FDG-avid tumor burden, and the maximum distance between the largest lesion and any other lesion quantifies the dissemination of the tumor, and both factors are inversely related to overall survival (OS) and progression-survival (PFS) in DLBCL. In this study, we explored prognostic impact of FDG PET/CT radiomics features in newly diagnosed IVLBCL. [Methods] We retrospectively evaluated patients with histopathologically proven newly diagnosed IVLBCL according to the 2008 World Health Organization criteria between January 2010 through February 2022 from hospitals participating in the North Japan Hematology Study Group (NJHSG). Quantitative analysis of all FDG PET/CT was performed using Metavol (Hokkaido University, Sapporo, Japan), with exclusion of physiological accumulation. MTV was defined as the volume of lymphoma lesions visualized on PET/CT scans with standardized uptake value (SUV) peaks greater than or equal to hepatic SUV peak. To quantify the dissemination of the tumor, we calculated the number of the body areas (head-neck, chest, abdomen, upper-extremities, lower-extremities) having BM lesions with a higher SUV peak than liver SUV peak. Patients with BM lesions spread across 4-5 areas were defined as cases having widely disseminated lesions (WDL). Progression-free survival (PFS) was defined as the time from the start of treatment to the first of progression, relapse, or death from any cause. Patients undergoing upfront autologous stem-cell transplantation (auto-SCT) were censored at the time of transplantation. [Results] We identified fifty-two IVLBCL patients (age 30-89 years, median 65 years; 57.7% male) who performed FDG PET/CT at diagnosis. Twenty-eight patients (54.9%) had poor performance status (ECOG PS ≥2), and 49 patients (94.2%) showed stage III-IV. Forty-nine patients (94.2%) had 2 or more extranodal involvement. Elevated serum LDH level was observed in 49 cases (94.2%) (median, 623.5 IU/L; range, 147-4907 IU/L). The median serum levels of soluble IL-2 receptor (sIL-2R), ferritin, and C-reactive protein (CRP) were 4496.5 IU/mL (range, 321-19900 IU/mL), 655.15 ng/mL (range, 50-6657 ng/mL), and 6.905 mg/dL (range, 0.11-25.86 mg/dL), respectively. The median MTV was 415.527 cm 3 (range, 0-3000 cm 3). ROC analysis revealed that the cutoff value of the MTV &amp;gt; 388 cm 3 could predict progression events within 2 years. Area under curve was 0.71 (95% CI: 0.564-0.857). Twenty-nine patients (55.8%) were classified as having MTV &amp;gt; 388 cm 3 (high MTV). Twenty-three patients (44.2%) had WDL. Fifty-one (98.1%) patients received chemotherapy, all of which was rituximab-containing regimen. Three patients (5.8%) received upfront auto-SCT. Median follow-up duration was 48.8 months (range, 5.8-140.5 months) in survivors. Age, LDH level, number of extranodal lesions, clinical stage, and PS did not affect OS and PFS. Although the high MTV and WDL did not significantly affect OS ( P = 0.095, P = 0.173, respectively; Log-rank), these findings did correlate significantly with poor PFS ( P = 0.011, P = 0.042, respectively; Log-rank). Thus, we defined a high MTV or a WDL as poor PET findings. Thirty-two patients had poor PET findings. Importantly, patients with poor PET findings showed significantly poor OS and PFS compared to patients without (2-year OS: 71.2% vs. 93.3%, P = 0.015, 2-year PFS: 45.5% vs. 93.8%, P = 0.001; Log-rank, Fig. 1, 2). [Conclusion] This is the largest study to assess prognostic impact of FDG PET/CT radiomics features on newly diagnosed IVLBCL. Poor PET findings were significantly correlated with worse OS and PFS.

Decreased lymphoma-related deaths and improved long-term relative survival with radiotherapy for early-stage diffuse large B-cell lymphoma in the rituximab era

BackgroundWe aimed to investigate the incidence of lymphoma-related death (LRD) and the long-term net survival benefit of radiotherapy (RT) for early-stage diffuse large B-cell lymphoma (DLBCL) in the rituximab era. Methods10,841 adults diagnosed with early-stage DLBCL between 2002–2015 were retrospectively analyzed using data from the Surveillance, Epidemiology, and End Results database. Primary therapy was categorized into combined-modality treatment (CMT, n = 3,631) and chemotherapy alone (n = 7,210). Competing risk analysis was used to evaluate the cumulative incidence of mortality. Inverse probability of treatment weighting (IPTW) was used to balance groups. The net survival benefit of RT was estimated through relative survival (RS), standardized mortality ratio (SMR), and transformed Cox regression, while controlling for background mortality. ResultsPatients initially treated with CMT had a lower cumulative incidence of LRD compared to those who received chemotherapy alone (HR 0.63, 95%CI: 0.57–0.69; P < 0.001). The 10-year overall survival (OS), RS, and SMR for CMT were 66.1%, 85.0%, and 1.71 respectively, which were significantly better than those for chemotherapy alone (53.0%; 69.8%; 2.62; all P < 0.001). IPTW and multivariable analysis revealed that the addition of RT led to better OS (HR 0.67, 95%CI: 0.62–0.71; P < 0.001) and RS (HR 0.69, 95%CI: 0.65–0.74; P < 0.001). Moreover, compared with chemotherapy alone, the benefit of OS and RS for CMT increased over time within 10 years of diagnosis. ConclusionRT reduced LRD and improved the long-term net survival in early-stage DLBCL in the rituximab era. Further prospective studies are warranted to assess the specific patient population that would benefit the most from consolidative RT in early-stage DLBCL.

Prediction and prognostic potential of NR3C1 gene expression level in DLBCL patients

ABSTRACTObjective: Diffuse Large B-Cell Lymphoma (DLBCL) is a common and frequently occurring subtype of Non-Hodgkin Lymphoma (NHL). The effective treatment and prognosis of DLBCL are still urgently needed to be explored. This article aims to shed light on the connection between DLBCL survival and NR3C1 expression levels. Methods: First, we divided the 952 DLBCL patients into an NR3C1 high-expression group and an NR3C1 low-expression group and compared the baseline characteristics of the two groups. Second, we used multivariate analysis to predict the dependent variable for age, pathology, ECOG score, lactate dehydrogenase (LDH) ratio, and NR3C1 expression level. Finally, we analyzed the progression-free survival (PFS) and overall survival rate (OS) of DLBCL patients with high or low NR3C1 expression. Results: DLBCL patients with high NR3C1 expression had a better prognosis than those with low NR3C1 expression (OS, P < 0.0001). In DLBCL patients of CHOP therapy, high NR3C1 expression was associated with a good survival prognosis in OS (OS, P = 0.028). Conclusion: In multivariate analysis, NR3C1 high expression was an independent prognostic factor that predicted a longer OS of DLBCL (OS, P = 0.0003). NR3C1 is considered an independent predictor of DLBCL patients and can be used as a biomarker for the prognosis of DLBCL.

Lower PTEN may be associated with CD8+ T cell exhaustion in diffuse large B-cell lymphoma

Initially discovered in chronic viral infection and then extended to tumor, 'T-cell exhaustion' is a broad term describing the response of T cells to chronic antigen stimulation. By definition, whether T-cell exhaustion occurs in diffuse large B-cell lymphoma (DLBCL) remains largely unknown because little has been described. Here, the immune-suppressing checkpoint molecules involved in T-cell exhaustion, including PD-1, PD-L1, TIM-3 and TIGIT, whose expression levels were analyzed in DLBCL, were retrieved from the GEPIA database. Compared with the normal control, CD8A, TNFA, IFNG and GZMA were markedly elevated in DLBCL, indicating that infiltrated CD8+ T cells predominate in DLBCL. Meanwhile, inhibitory immune checkpoints, such as PD-1, PD-L1, TIGIT and TIM-3 were drastically higher in DLBCL. PTEN, WNT2 and DKK3 expression were also appraised. It was revealed that PTEN was lower in DLBCL, without being statistically significant. In contrast with PTEN, DKK3 and WNT2 were shown to be pronouncedly higher in DLBCL relative to the normal control. Prognostically, only TIGIT was found to be associated with overall survival in DLBCL. Collectively, all the data we curetted from the GEPIA and TIMER 2.0 databases explicitly indicate that CD8+ T cell exhaustion took place, which may be linked with lower PTEN in DLBCL. To the best of our knowledge, this is the first bioinformatic report explicitly proposing that CD8+ T cell exhaustion occurs in DLBCL, which may be associated with lower PTEN.

EXPLORING DIFFERENTIAL MUTATIONAL DISTRIBUTION BY TARGETED SEQUENCING IN A R‐CHOP TREATED COHORT OF DIFFUSE LARGE B‐CELL LYMPHOMA PATIENTS

Background: Diffuse large B-cell lymphoma (DLBCL) harbors striking clinical and molecular heterogeneity, which continues to encumber risk stratification with a consequently high rate of refractory/relapsed patients (pts). Though the genomic landscape of DLBCL has been comprehensively uncovered, the search for eligible molecular biomarkers continues. Exploratively, we set out to retrospectively evaluate differential mutational distributions in a DLBCL cohort using targeted sequencing. Patients and method: A cohort of 105 de novo DLBCL pts, uniformly treated with first-line R-CHOP, were studied. Thirty pts relapsed, of which 70% had early relapses (<2 years after completing therapy), while 30% had late relapses (≥2 years). Targeted next-generation sequencing (90 gene panel) was applied on archival paraffin-embedded diagnostic samples and, if available, first relapse samples. Results: In total, 118 samples were sequenced, each displaying a median of 12 (range 1–46) putative somatic protein-coding mutations. Germinal center B-cell (GCB) DLBCL (n = 60) displayed a differential somatic signature with enrichment of EZH2, TNFRSF14, BCL2, ACTB, SOCS1, and FAS mutations (Fisher's Exact, p < 10-4), whereas mutations in CDKN2A, PRDM1, MYD88, and CD79B were significantly enriched in non-GCB pts (p < 10-4, n = 44). Survival analysis confirmed inferior overall survival (OS) in non-GCB DLBCL (5-year OS, log-rank (Mantel-Cox), p = 0.01, hazard ratio (HR) 1.7). Pts with double-hit biology (DHB), defined as either MYC and BCL2 and/or BCL6 fluorescent in situ hybridized positive and/or with MYC and BCL2 immunohistochemical double expression (n = 32), did not display a significant mutational signature or differential OS. However, a subgroup of DHB pts characterized by mutations in MYC, PRDM1, or IRF4 (n = 15) had markedly inferior OS (5-year OS, log-rank, p = 0.0003, HR 3.34) compared to negative non-DHB pts (n = 55) or negative non-DHB/DHB pts (p = 0.0017, HR 2.7, n = 72). Noticeably, ACTB mutations were only present in non-DHB pts. The mutational distribution at diagnosis did not differ between pts with relapse and in sustained remission. However, in the relapse group, GNA13 mutation at diagnosis was more frequent in early relapses than late relapses (43% vs. 0%, Fisher’s Exact without correction, p = 0.03). The presence of GNA13 in the entire cohort showed a trend towards inferior survival. Conclusion: Differential distribution with significant mutational signatures were expectedly found in GCB versus non-GCB, but not in subgroups with DHB or relapse. Interestingly, mutations in MYC, PRDM1 or IRF4 identified a distinct subset of DHB pts with inferior survival. Furthermore, GNA13 mutations were significantly enriched in early relapse pts. These preliminary findings call for validation of GNA13 as a potential marker of early relapse as well as the prognostic value of MYC, PRDM1 and IRF4 mutations in DHB DLBCL. The research was funded by: The Danish Cancer Society and The Vissing Foundation Keywords: aggressive B-cell non-Hodgkin lymphoma, genomics, epigenomics, and other -omics, tumor biology and heterogeneity Conflicts of interests pertinent to the abstract T. S. Larsen Consultant or advisory role Roche, Gilead, Novartis, Celgene/BMS Research funding: Genentech