Diffuse Large B-cell Lymphoma Group Research Articles

Comparative Analysis of the Clinical Manifestations of Sinonasal Natural Killer T-Cell Lymphoma and Diffuse Large B-Cell Lymphoma

Background and Objectives: Natural killer T-cell lymphoma (NKTCL) and diffuse large B-cell lymphoma (DLBCL) are the two most prevalent subtypes of lymphoma in the sinonasal region. Accurately differentiating between sinonasal DLBCL and NKTCL is crucial for determining the appropriate treatment and prognosis. The present study compared the clinical characteristics of these two conditions.Methods: We conducted a retrospective review of 173 patients diagnosed with sinonasal lymphoma at a single institute between 2004 and 2017. This review included only patients with DLBCL and NKTCL who had more than 6 months of follow-up records. We analyzed patient data encompassing clinical characteristics, pathologic findings, radiologic findings, treatment modalities, recurrence, and survival.Results: Among the patients analyzed, 117 patients were diagnosed with NKTCL and 45 with DLBCL. Endoscopic evaluation revealed a significantly higher incidence of crusting (p<0.001) and necrotic lesions (p=0.001) in patients with NKTCL, whereas polypoid masses were more commonly observed in patients with DLBCL (p<0.001). Computed tomography (CT) scans indicated no significant differences in bilaterality or bone destruction between the two groups. The DLBCL group exhibited a higher rate of concurrent lymph node or organ involvement than the NKTCL group (p<0.001). The 5-year overall survival rate was 67.4% for DLBCL and 69.1% for NKTCL, with no significant difference between the two.Conclusion: Clinical differences exist between sinonasal DLBCL and NKTCL in terms of endoscopic and CT findings. These distinct characteristics can aid in distinguishing between the two types of sinonasal lymphoma during clinical diagnosis.

Cuproptosis-related lncRNA signature as a prognostic tool and therapeutic target in diffuse large B cell lymphoma

Cuproptosis is a newly defined form of programmed cell death that relies on mitochondria respiration. Long noncoding RNAs (lncRNAs) play crucial roles in tumorigenesis and metastasis. However, whether cuproptosis-related lncRNAs are involved in the pathogenesis of diffuse large B cell lymphoma (DLBCL) remains unclear. This study aimed to identify the prognostic signatures of cuproptosis-related lncRNAs in DLBCL and investigate their potential molecular functions. RNA-Seq data and clinical information for DLBCL were collected from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). Cuproptosis-related lncRNAs were screened out through Pearson correlation analysis. Utilizing univariate Cox, least absolute shrinkage and selection operator (Lasso) and multivariate Cox regression analysis, we identified seven cuproptosis-related lncRNAs and developed a risk prediction model to evaluate its prognostic value across multiple groups. GO and KEGG functional analyses, single-sample GSEA (ssGSEA), and the ESTIMATE algorithm were used to analyze the mechanisms and immune status between the different risk groups. Additionally, drug sensitivity analysis identified drugs with potential efficacy in DLBCL. Finally, the protein–protein interaction (PPI) network were constructed based on the weighted gene co-expression network analysis (WGCNA). We identified a set of seven cuproptosis-related lncRNAs including LINC00294, RNF139-AS1, LINC00654, WWC2-AS2, LINC00661, LINC01165 and LINC01398, based on which we constructed a risk model for DLBCL. The high-risk group was associated with shorter survival time than the low-risk group, and the signature-based risk score demonstrated superior prognostic ability for DLBCL patients compared to traditional clinical features. By analyzing the immune landscapes between two groups, we found that immunosuppressive cell types were significantly increased in high-risk DLBCL group. Moreover, functional enrichment analysis highlighted the association of differentially expressed genes with metabolic, inflammatory and immune-related pathways in DLBCL patients. We also found that the high-risk group showed more sensitivity to vinorelbine and pyrimethamine. A cuproptosis-related lncRNA signature was established to predict the prognosis and provide insights into potential therapeutic strategies for DLBCL patients.

HLA variations in patients with diffuse large B-cell lymphoma and association with disease risk and prognosis: a case-control study.

Human leukocyte antigen (HLA) polymorphisms have been associated with the development of various autoimmune diseases, as well as malignant neoplasms. Non-Hodgkin lymphomas (NHLs) are a heterogenous group of lymphoid malignancies in which a genetic substrate has been established and is deemed to play a crucial role in disease pathogenesis. This study aimed to identify whether variations in the HLA gene region were associated with diffuse large B-cell lymphoma (DLBCL) risk and prognosis. We defined HLA class I (HLA-A, HLA-B, HLA-C) and class II (HLA-DRB1, HLA-DQB1) alleles in 60 patients with DLBCL and compared the results to those found by 236 healthy adult donors from the bone marrow bank of Northern Greece. HLA typing was performed by two molecular methods, Sequence - Specific Oligonucleotide HLA typing (SSO) and Sequence - Specific Primer HLA typing (SSP), from white blood cells recovered from peripheral blood. The phenotypic frequencies of HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 between patients and controls were compared with the 2-sided Fisher's exact test. Results with p-value <0.05 were considered statistically significant. Odds Ratios with 95% Confidence Intervals were calculated to further strengthen the results. The 2-sided Fisher's exact test was also applied to alleles found only in one of the two groups, while the odds ratios together with the confidence intervals were corrected with Haldane-Anscombe method. Among the studied HLA polymorphisms, the frequency HLA-C*12 allele was significantly lower in patients with DLBCL compared with control subjects (6.7% vs. 34.7%, OR = 0.16, 95% CI: 0.04-0.44). Frequency of HLA-B*39 was significantly lower in patients with DLBCL compared with controls, but due to the low frequency of this polymorphism in the studied population and small sample size, determinations regarding the significance of this findings were limited. Survival analysis revealed that the presence of HLA-C*12 was not associated with improved or worsened overall and progression-free survival. No statistically significant associations were observed in the phenotypic frequencies of HLA-A, HLA-DQB1, HLA-DRB1 and the rest of HLA-B alleles between the control and DLBCL groups. Collectively, our results provide valuable insight regarding the role of HLA variations on DLBCL risk. Further studies are required to consolidate our findings and ascertain the clinical implications of these genetic variations on DLBCL management and prognosis.

The Possible Role of Pathogens and Chronic Immune Stimulation in the Development of Diffuse Large B-Cell Lymphoma.

Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin lymphoma. The disease is very heterogeneous, with distinct genetic alterations in subtypes. The WHO 2022 5th edition classification identifies several minor groups of large B-cell lymphoma where the pathogenetic role of viruses (like EBV and HHV-8) is identified. Still, most cases fall into the group of DLBCL not otherwise specified (NOS). No review focuses only on this specific lymphoma type in the literature. The pathogenesis of this entity is still not fully understood, but several viruses and bacteria may have a role in the development of the disease. The authors review critical pathogenetic events in the development of DLBCL (NOS) and summarize the data available on several pathogenetic viruses and bacteria that have a proven or may have a potential role in the development of this lymphoma type. The possible role of B-cell receptor signaling in the microenvironment is also discussed. The causative role of the Epstein-Barr virus (EBV), human herpesvirus-8 (HHV-8), Hepatitis C virus (HCV), human immunodeficiency virus (HIV), Hepatitis B virus (HBV), and other viruses are explored. Bacterial infections, such as Helicobacter pylori, Campylobacter jejuni, Chlamydia psittaci, Borrelia burgdorferi, and other bacteria, are also reviewed.

Diverse B-cell tumors associated with t(14;19)(q32;q13)/IGH::BCL3 identified by G-banding and fluorescence in situ hybridization.

We characterized 5 B-cell tumors carrying t(14;19)(q32;q13) that creates the IGH::BCL3 fusion gene. The patients' ages ranged between 55 and 88 years. Two patients presented with progression or recurrence of B-cell chronic lymphocytic leukemia (B-CLL)/small lymphocytic lymphoma (SLL), two with diffuse large B-cell lymphoma (DLBCL) of non-germinal center B-like phenotype, and the remaining one with composite angioimmunoblastic T-cell lymphoma and Epstein-Barr virus-positive DLBCL. The presence of t(14;19)(q32;q13) was confirmed by fluorescence in situ hybridization (FISH), showing colocalization of 3' IGH and 3' BCL3 probes on der(14)t(14;19) and 5' BCL3 and 5' IGH probes on der(19)t(14;19). One B-CLL case had t(2;14)(p13;q32)/IGH::BCL11A, and 2 DLBCL cases had t(8;14)(q24;q32) or t(8;11;14)(q24;q11;q32), both of which generated IGH::MYC by FISH, and showed nuclear expression of MYC and BCL3 by immunohistochemistry. The IGH::BCL3 fusion gene was amplified by long-distance polymerase chain reaction in 2 B-CLL/SLL cases and the breakpoints occurred immediately 5' of BCL3 exon 1 and within the switch region associated with IGHA1. The 5 cases shared IGHV preferentially used in B-CLL cells, but the genes were unmutated in 2 B-CLL/SLL cases and significantly mutated in the remaining 3. B-cell tumors with t(14;19)(q32;q13) can be divided into B-CLL/SLL and DLBCL groups, and the anatomy of IGH::BCL3 in the latter may be different from that of the former.

Primary Thyroid Lymphoma: Clinical Factors Predicting the Possibility of Diffuse Large B-Cell Lymphoma.

Aims: Among primary thyroid lymphomas (PTLs), diffuse large B-cell lymphoma (DLBCL) has a poorer prognosis than other indolent lymphomas such as mucosa-associated lymphoid tissue (MALT) or follicular lymphoma (FL). However, the clinical differences between DLBCL and indolent lymphoma remain unclear. Therefore, this retrospective study on PTL was aimed at investigating the clinical differences between DLBCL and indolent lymphomas and identifying the factors differentiating DLBCL from indolent lymphomas. Materials and Methods: Medical records of 28 patients diagnosed with PTL and treated at our institution between 2005 and 2022 were retrospectively analyzed. Data on the following clinical variables were extracted: sex, age, symptoms (pain and dysphagia), ultrasonographic appearance patterns, the presence of airway stenosis on computed tomography and laryngeal endoscopy, blood test results, disease stage, and pathological diagnosis. Results: In all, 13 patients were histologically diagnosed with DLBCL, 12 with MALT lymphoma, and 3 with FL. Significant differences in disease-specific survival rates were evident between the DLBCL and indolent lymphoma groups (68.2 vs 100%, P = .043). High lactate dehydrogenase levels (>230 U/mL) and airway stenosis were observed only in patients with DLBCL. Multivariate analysis identified that the presence of a linear echoic strand pattern and the absence of an echoic nodular pattern on ultrasound were independently associated with DLBCL (P = .0497 and .012, respectively). Conclusion: DLBCL can cause airway stenosis. The linear echogenic strand pattern and the absence of a nodular pattern should be recognized as predictive factors of DLBCL.

Comparison of Efficacy and Side Effects of BTK Inhibitors for Different Duration As a Bridging Therapy before Anti-CD19-CAR T-Cell Therapy in Patients with R/R DLBCL

Background: BTK inhibitors (BTKi)was used for bridge therapy before anti-CD19-CAR T-cell therapy in patients with relapsed/refractory (R/R) diffuse large B-cell lymphoma (DLBCL) could improve the efficacy of anti-CD19-CAR T-cell therapy. However, the duration of BTKi bridging prior to anti-CD19-CAR T-cell therapy has not been clear. Methods: This was a single-center, investigator-initiated clinical study involving patients with R/R DLBCL, which has at least one high risk factor for poor prognosis. All these R/R DLBCL patients received BTKi (Ibrutinib or Zanubrutinib) bridging therapy for one to four months before their anti-CD19-CAR T-cell therapy. After CAR-T cell infusion, BTKi therapy was continued. All patients were divided into two groups based on the duration of BTK inhibitors bridging therapy: ≥2 mons group and&amp;lt;2 mons group. We observed the overall response rate (ORR) and overall survival (OS), CAR-T cell amplification, expression of programmed cell death 1 (PD-1), cytokines and adverse events. Results: In our study, all the thirty-one patients with R/R DLBCL had at least one high risk factor for poor prognosis before their anti-CD19-CAR T-cell therapy. Except for the primary lines, proportion of Richter transformation patients, there were no differences in age, sex, stage, International Prognostic Index scores between the ≥2 mons group and &amp;lt;2 mons group. Two patients failed to receive CAR-T cell infusion due to disease progression while receiving BTKi bridging therapy. Sixteen patients (55.2%) obtained CR, six patients (20.7%) obtained PR, six patients (20.7%) obtained SD and one patient (3.4%) obtained PD only. The ORR of this study was 75.9% (22/29). The median duration of BTKi bridging therapy in patients with ORR was 63 days, which was higher than that of in patients with SD or PD, which was 28 days. The peaks of CAR-T cells were higher in ≥2 mons group than that of in &amp;lt;2 mons group, higher in ≥2 mons+ Richter group than that of in &amp;lt;2 mons+ DLBCL group, but there was no difference of the peaks between the DLBCL group and Richter group. The ≥grade 2 of CRS was found in ten patients (71.61%) in ≥2 mons group, and only three patients (20.00%) in &amp;lt;2 mons group. The ≥grade 2 of CRS was found in five patients (35.71%) in DLBCL group, and eight patients (53.33%,) in Richter group. The ≥grade 2 of CRS was found in six patients (66.67%) in ≥2 mons+ Richter group, and only one patient (11.11%) in &amp;lt;2 mons+ DLBCL group. The results were similar for ICANS and CRS in our study. The ≥ grade 2 of neutropenia, anemia and thrombocytopenia were found in eleven patients (78.57%), seven patients (50.00%) and five patients (35.71%) in ≥2 mons group, and one patient (6.67%), one patient (6.67%) and one patient (6.67%) in &amp;lt;2 mons group. None of the patients died of bacterial infections or invasive fungal diseases. Conclusion: The duration of BTKi bridging therapy before anti-CD19-CAR T-cell therapy affects the efficacy and side effects of R/R DLBCL patients, especially hematological toxicity. R/R DLBCL patients with higher primary lines or Richter transformation could be improved the efficacy of anti-CD19-CAR T-cell therapy by prolonging BTKi bridging therapy (≥2 mons). However, there were no serious adverse results due to hematological toxicity.

HIV-Related Diffuse Large B Cell Lymphoma Influences on T Cell Exhaustion

BACKGROUD HIV infection leads to T cell dysfunction, which triggers increased incidence of diffuse large B cell lymphoma (DLBCL). It has been shown that PD-1 inhibitors can better restore T cell function and effectively treat HIV-related lymphoma. However, the specific mechanism on the treatment of HIV-related DLBCL patients with PD-1 inhibitors is ambiguous, and related studies on the expression of PD-1 on T cells have not been reported. This study aimed to explore the regulatory role of altered immune function status in the microenvironment of HIV-related DLBCL and to analyze hyper-exhausted T cell subsets. METHODS Three patients with DLBCL, two patients with HIV and 11 patients with HIV-related DLBCL who were treated from September 2021 to December 2022 at the Chongqing University Cancer Hospital of were included. Single cell sequencing was used to analyze immune cell and tumor cell related genes in patients'lymph node tissues. InferCNV was used to distinguish normal and malignant B lymphocytes. Gene set enrichment analysis (GSEA) was used to find the expression of exhaustion genes and related signaling pathways in immune cells. And relevant exhausted immune cells were analyzed and validated by flow cytometry. RESULTS Compared to the HIV and DLBCL groups, the HIV-related DLBCL group had a higher level of expression of the gene PDCD1, a higher level of expression of CD23(FCER2) in normal B lymphocytes, and a greater difference in malignant B lymphocytes, with a lower percentage of TCF1-positive exhausted T cells and a higher level of CXCL13-positive exhausted T cells (Fig.1-3). Compared with the HIV group, the percentage of follicular helper T cells (T fh) was lower in the HIV-related DLBCL group. The results of the Pseudotime analysis showed two main developing directions of TCF1-positive exhausted T cells (Fig.4). The state2 was primarily the direction of TCF1-positive exhausted T cell development in the HIV group and the HIV-related DLBCL group, and the state3 was primarily the direction of TCF1-positive exhausted T cell development in the DLBCL group and the HIV-related DLBCL group. CONCLUSION Patients with HIV-related DLBCL are more at risk of T cell exhaustion. In this study, we found that HIV-related DLBCL induced T cell exhaustion has different states and shows two differentiation directions, and state2 may correlate with HIV-induced T cell exhaustion. T cell exhaustion states may be used for disease status and clinical efficacy assessment of HIV-related DLBCL.

Prediction of Survival in Diffuse Large B Cell Lymphoma According to Immunohistochemistry

BACKGROUND: The introduction of chemoimmunotherapy in the treatment of diffuse large B cell lymphoma (DLBCL) dramatically improved the outcomes compared to chemotherapy alone. However, a significant part of the patients become refractory and eventually relapse. AIM: The purpose of this study is to analyze the survival differences between different groups of DLBCL according to Han’s algorithm. MATERIALS AND METHODS: We will study the medical records of 244 patients treated with RCHOP as first-line therapy who are diagnosed with de novo DLBCL in a cohort of 10 years in the University Clinical Center of Kosovo. According to immunohistochemical markers, the patients will be classified into two major groups, germinal center B cell-like (GCB) and non-germinal center B cell-like (non-GCB) subtypes, and five subgroups (Group 1, Group 2, Group 3 or double positive, Group 4, and Group 5 or triple negative [TN]). RESULTS: The patients in the GCB group have better progression-free survival (PFS) and overall survival (OS) than the non-GCB group. Despite this, double positive (DP) (CD10+MUM1+) and TN (CD10−Bcl6−MUM−) subgroups showed different PFS and OS to the same cell-of-origin group. The DP group showed similar OS and PFS with the non-GCB group, whereas the TN group showed similar OS and PFS with the GCB group. CONCLUSION: These factors will provide valuable knowledge for predicting the prognosis and redirecting the choice of treatment for different groups of DLBCL.

Value of GCET1, HGAL (GCET2), and LMO2 in the Determination of Germinal Center Phenotype in Diffuse Large B-cell Lymphoma

Diffuse large B-cell lymphoma (DLBCL) is a biologically heterogeneous disease that is classified into germinal center B-cell (GCB) and non-GCB subtypes, which are prognostically different. The Hans algorithm is the most widely used tool based on CD10, BCL6, and MUM1 expression, but some cases with the non-GCB phenotype are still known to be misclassified. In this study, we investigate the extent to which GCET1, HGAL, and LMO2 protein expressions reflect GCB phenotype together with their roles in determining the GCB phenotype of DLBCL and their contributions to the performance of the Hans algorithm. Sixty-five cases of DLBCL-not otherwise specified, 40 cases of follicular lymphoma (FL), and 19 non-GC-derived lymphoma cases were included in this study. The DLBCL cases were grouped as CD10+ (Group A) or only MUM1+ (Group B), and the remaining cases constituted the intermediate group (Group C). GCET1, HGAL, and LMO2 expressions were evaluated. In the FL group, GCET1, HGAL, and LMO2 were positive in 85%, 77.5%, and 100% of the cases, respectively. Among the non-GC-derived lymphoma cases, all three markers were negative in cases of small lymphocytic lymphoma, plasmablastic lymphoma, peripheral T-cell lymphoma, and anaplastic large cell lymphoma. GCET1 and HGAL were negative in cases of marginal zone lymphoma (MZL) and mantle cell lymphoma (MCL). Two of the 3 MZL and 2 of the 4 MCL cases were positive for LMO2. In the DLBCL group, the number of cases with GCET1, HGAL, and LMO2 positivity was 18 (90%), 17 (85%), and 20 (100%), respectively, in Group A and 0 (0%), 2 (13.3%), and 2 (13.3%), respectively, in Group B. Considering these rates, when the cases in the intermediate group were evaluated, it was concluded that 13 cases typed as non-GCB according to the Hans algorithm may have the GCB phenotype. GCET1, HGAL, and LMO2 are highly sensitive markers for determining the germinal center cell phenotype and can increase the accuracy of the subclassification of DLBCL cases, especially for cases that are negative for CD10.

Improved efficacy with early tocilizumab in the prophylaxis and treatment of cytokine release syndrome of chimeric antigen receptor T cell (CAR-T) therapy for diffuse large B-cell lymphoma than acute lymphoblastic leukemia

BackgroundTocilizumab is a well-practiced strategy to manage cytokine release syndrome (CRS) after chimeric antigen receptor T cell (CAR-T) therapy. However, varied efficiency in CRS mitigation by tocilizumab has been reported to highlight affecting variables such as tumor types and timing of the administration. ObjectiveThe objective of this study is to identify different curative effect between diffuse large B-cell lymphoma (DLBCL) and acute lymphoblastic leukemia (ALL) by early use of tocilizumab for prophylaxis and treatment of CRS. MethodsBy retrospectively summarizing our institution's experience with a CAR-T clinical trial targeting CD19, 11 cases were analyzed: 5 DLBCL, and 6 ALL. The two groups were compared with patient characteristics, CRS information and clinical outcomes. 3 patients were pretreated with tocilizumab for prophylaxis and the rest were treated with tocilizumab at CRS diagnosis. ResultsCRS occurred in 81.8% of patients (9/11), grade 3 or higher occurred in 55.6% of the CRS patients (5/9). The two group were similar in patient characteristics, CAR-T and CRS profile. However, tocilizumab produced much better efficacy against CRS in DLBCL group compared with ALL group (80% versus 16.7%, P ​= ​0.08). ConclusionsDespite the statistical was non-significant, possibly due to small case pool and bias was unavoidable, our analysis suggested that early use of tocilizumab was more effective for DLBCL than ALL in the prophylaxis and treatment of CRS. A treatment algorithm for management of CRS with regard to DLBCL and ALL is proposed, which may extend to other local or systemic malignancies and warrant further investigation.

End-of-Life Care of Acute Myeloid Leukemia Compared with Aggressive lymphoma in Patients Who Are Eligible for Intensive Chemotherapy: An Observational Study in a Japanese Community Hospital.

Patients with hematological malignancies (HMs) are reported to receive more aggressive care at the end of life (EOL) than patients with solid tumors. However, the reasons behind this occurrence are not fully understood. To examine whether the care at EOL for HMs is mainly because of the disease characteristics or hematologists' attitudes and systems of care, we compared the EOL care of patients with acute myeloid leukemia (AML) and diffuse large B cell lymphoma (DLBCL). We retrospectively analyzed the EOL care of patients with AML and DLBCL younger than 80 years who were receiving combination chemotherapy at a city hospital in Japan. Fifty-nine patients with AML and 65 with DLBCL were included. Those with AML received chemotherapy more often within their last 30 days (48% vs. 19%, p < 0.001) and 14 days (37% vs. 1.5%, p < 0.001) of life, and consulted the palliative team less frequently (5.3% vs. 29%, p < 0.001). In the last 3 years, the mortality rate in hematological wards decreased from 74% to 29% in the DLBCL group, but only from 95% to 90% in the AML group. In multivariate analysis, AML (odds ratio [OR] 0.065) and death before 2018 (OR, 0.077) were significant factors associated with reduced referrals to specialized palliative teams. Patients with AML tend to have lesser access to specialized palliative care and fewer options for their place of death than those with DLBCL. Detailed EOL care plans are needed for these patients, considering the characteristics of the disease.

T-cell immunoglobulin mucin molecule-3, transformation growth factor β, and chemokine-12 and the prognostic status of diffuse large B-cell lymphoma.

The effects of T-cell immunoglobulin mucin molecule-3 (Tim-3), transforming growth factor β (TGF-β), and chemokine-12 (CXCL12) expression on the prognosis of patients with diffuse large B-cell lymphoma (DLBCL) have not been elucidated. To examine the correlation between Tim-3, TGF-β and CXCL12 expression and DLBCL prognosis. Lymph node tissues of 97 patients with DLBCL and 93 normal-response hyperplastic lymph node tissues treated from January 2017 to May 2019 were selected as the DLBCL and control groups, respectively. The expression of Tim-3, TGF-β, and CXCL12 was detected immunohistochemically. Patients were followed up for 3 years, and progression-free survival was recorded. Cox multifactorial analysis was performed to analyze the risk factors for poor prognosis. The positive expression rates of Tim-3, TGF-β, and CXCL12 were higher in DLBCL tissues than in non-cancerous (control) tissues (P < 0.05). One-year post-surgery, the positive expression rates of Tim-3, TGF-β, and CXCL12 were higher in patients with effective treatment than in those with ineffective treatment (P < 0.05). The 3-year progression-free survival of 97 patients with DLBCL was 67.01% (65/97). Univariate analysis revealed that clinical stage, bone marrow infiltration, International Prognostic Index (IPI) score, Tim-3 positivity, TGF-β positivity, and CXCL12 positivity were associated with poor prognosis (P < 0.05). Multivariate Cox regression analysis demonstrated that clinical stage III-IV, bone marrow infiltration, mediate-to-high-risk IPI scores, Tim-3 positivity, TGF-β positivity, and CXCL12 positivity were independent risk factors affecting prognosis (P < 0.05). DLBCL tissues exhibit high positive expression of Tim-3, TGF-β, and CXCL12, and a high expression of all three indicates a poor prognosis.

Untargeted Metabolomics of Cerebrospinal Fluid in Patients with Primary Central Nervous System Lymphoma

Primary central nervous system lymphoma (PCNSL) is characterized by a high degree of malignancy and poor prognosis. The current diagnostic techniques include cerebrospinal fluid (CSF) metabolomics examination, imaging examination, needle biopsy, etc., but each examination has certain limitations, and it is difficult for the early identification and diagnosis of the disease. In terms of treatment, relapse and refractory, neurotoxicity and drug resistance are still the main problems currently faced. Metabolomics can measure the composition of metabolites and the dynamic changes of metabolites in cell models, tissues, organs or the whole organism, and has been widely used in disease diagnosis, efficacy evaluation, nutrition and other fields. Ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry (UPLC-HRMS/ technology) has been be used to observe the metabolites changes in CSF. Here, we aimed to explore the application value of metabolomics in the diagnosis and prognosis evaluation of PCNSL. From January 2021 to December 2021, CSF samples from 37 DLBCL patients were collected in Shengli Oilfield Central Hospital and Shandong Cancer Hospital, including 20 male patients and 17 female patients. These patients fall into 4 groups based on clinical presentations: 12 patients in PCNSL group (CNS group), 5 patients in primary testicular diffuse large B-cell lymphoma (DLBCL) group (TE group), 10 patients in Germinal center B (GCB) cell-like DLBCL without central nervous system infiltration (GCB group), and 10 patients in non-GCB DLBCL group without central nervous system infiltration (non-GCB group). The CSF samples were collected before clinical treatment, and the samples were analyzed with UPLC-HRMS/technology. For data analysis, principal component analysis (PCA) was carried out on the obtained components, and the partial least squares discriminant analysis model (PLS-DA) was used for diversification analysis. Analytes with P<0.05 and VIP>1 were considered differential metabolites. The Venn diagram was used to narrow the range of metabolites. By using MetaboAnalyst 5.0 software, pathway enrichment analysis was performed on the identified differential metabolites in CSF, and the corresponding P value was obtained. The metabolic pathways that have a significant effect in CNS group vs TE group, CNS group vs GCB group and GCB group vs non-GCB group were glycerophospholipid metabolism and sphingolipid metabolism. The metabolic pathway that has a significant effect in CNS group vs non-GCB group and TE group vs non-GCB group was glycerophospholipid metabolism. Fatty acid degradation and fatty acid elongation were the significant effects metabolic pathway in TE group vs non-GCB group. We applied Venn diagrams to narrow the range of differential metabolites. There were 47 differential metabolites in the CSF of patients in the PCNSL group vs the non-PCNSL group (Figure 1). Compared with the non-PCNSL group, 33 metabolites such as 3-Hydroxyisovalerylcarnitine, Hesperetin 3'-O-sulfate, and Arteether were up-regulated and 14 metabolites such as Pentaglutamyl folate, Calcipotriol and N-Hexadecanoylpyrrolidine were down-regulated in the CSF of patients in the PCNSL group. In the CNS infiltration high-risk group and the CNS infiltration low-risk group, there were 16 differential metabolites (Figure 2). Compared with the patients in the CNS infiltration low-risk group, 8 metabolites such as 3-Allylphenol sulfate, p-Aminobenzoic acid and 2,3,4-Trichlorophenol were up-regulated and 8 metabolites such as Palmitaldehyde, Carboxamide and Rioprostil were down-regulated in the CNS infiltration high-risk group. In conclusion, glycerophospholipid, sphingolipid, and fatty acid metabolites were the three most important abnormal lipid metabolites existed in PCNSL. This finding may have some value in the development of diagnosis and prognostic tools for evaluating CSF in patients with PCNSL. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

BCL6 Rearranged Indolent B-Cell Lymphomas Show Mutational Profile Similar to Marginal Zone Lymphomas

Introduction Follicular lymphoma (FL) is the most common indolent non-Hodgkin lymphoma. It is a neoplasm of germinal center (GC) B cells with hallmark BCL2 rearrangements (BCL2-R). A distinct subgroup of indolent B cell lymphomas (iBCL), histologically revealing a follicular growth pattern but lacking BCL2-R and with BCL6 rearrangements (BCL6-R) has been described. Historically they have been classified as FL and included under the FL entity in the WHO classification. However, recent studies on diffuse Large B cell lymphomas (DLBCL) (1, 2) have described BCL6-R in ~20% of DLBCL, with a subset of cases showing concurrent gene mutations commonly found in marginal zone lymphomas (MZL) rather than those characteristic of FL or GCB DLBCL. Systematic studies elucidating the clinicopathologic and genetic features of the BCL6-R iBCL are currently lacking thus generating controversy over their appropriate classification and management. Methods The MSKCC cytogenetic lab database was searched from 01/2006 to 07/2021 and a cohort of 131 iBCL with BCL6-R (71 with BCL6-R only and 60 with both BCL6-R & BCL2-R) was obtained (Table 1). The clinical, morphologic, immunophenotypic, and molecular results (50 cases had molecular data) were reviewed. These were compared to 70 BCL2-R conventional FL (cFL; all having molecular data) and 54 BCL2-R and BCL6-R negative iBCL (17 had molecular data). ResultsBCL6-R iBCL showed distinct histological, immunophenotypic, genetic and clinical features. They showed a significantly higher number of Grade 3 FL diagnosis and CD10 negative phenotype. The most commonly mutated genes in BCL6-R only group were KMT2D (48%) and TNFAIP3 (40%); in the BCL6-R & BCL2-R group, KMT2D (72%) and BCL2 (56%); and in the cFL group KMT2D (83%), BCL2 (81%) and CREBBP (66%). Unsupervised clustering (K-Means) performed to cluster the samples based on the mutational status of about 50 genes revealed that majority of the BCL6-R only cases clustered separately from cFLs. They clustered with majority of BCL2-R and BCL6-R negative cases and revealed frequent mutations in TNFAIP3, BCL10, B2M, NOTCH2, SPEN (Fig 1). Majority (2/3) of the BCL6-R & BCL2-R cases clustered with cFL and showed mutations commonly seen in FL (Fig 1). The overall survival (OS) was significantly inferior in the BCL6-R cases (BCL6-R only as well as both BCL6-R & BCL2-R cases) in comparison to cFL even though progression free survival was shortest for cFL. ConclusionBCL6-R only iBCL are biologically distinct, cluster separately from cFL based on mutational profile, and demonstrate an MZL-like mutational profile similar to that is described for the BN2 group of DLBCL (1). These findings suggest that they are a distinct entity, different from cFL and possibly precursor lesions for BN2 DLBCL. In contrast, majority of the BCL6-R & BCL2-R cases cluster with cFL and show mutations commonly seen in FL. Finally, presence of BCL6-R was associated with a worse OS.

The multilobated morphology is still a better prognosis factor of diffuse large B-cell lymphoma in the R-CHOP era.

Diffuse large B-cell lymphoma (DLBCL) is the most common type of B-cell lymphoma. Although the multilobated subtype of DLBCL has been observed since the 1970s, little is known about the clinical significance of this unique variant in the era of rituximab, cyclophosphamide, hydroxydaunorubicin, oncovin, prednisone/prednisolone (R-CHOP) therapy. In this study, the retrospective clinicopathological analysis of 312 patients diagnosed with DLBCL showed that the multilobated DLBCL group comprised 11% of the cases and was predominantly male (p = 0.027), achieved complete remission in the first therapy (p = 0.023), and exhibited germinal center B-cell phenotypes in the Hans algorithm (p = 0.025). The multilobated DLBCL groups had a better prognosis in overall survival (OS) and progression-free survival (PFS) than the non-multilobated DLBCL group (OS, p = 0.006; PFS, p = 0.010). In the multivariate Cox regression analyses for OS, independent prognosis factors were high soluble IL-2 receptor (p = 0.025), high risk of International Prognostic Index, and multilobated morphology (p = 0.031). The most characteristic copy number gains found in more than 50% of the cases were located at 1q, 3p, 10q, 12q, and 14q. Overall, the multilobated morphology in DLBCL exhibits a good outcome in the R-CHOP era.

CD5+ diffuse large B-cell lymphoma has heterogeneous clinical features and poor prognosis: a single-center retrospective study in China.

ObjectiveDe novo CD5-positive (CD5+) diffuse large B-cell lymphoma (DLBCL) has different clinical characteristics compared with CD5-negative (CD5−) DLBCL. However, few studies have been reported in Chinese cohorts. We investigated the clinical features and prognosis of patients with CD5+ DLBCL and summarized the related literature.MethodsData from 245 patients with newly diagnosed DLBCL were retrospectively assessed.ResultsThirty-one and 214 patients were diagnosed with CD5+ DLBCL or CD5− DLBCL, respectively. In the CD5+ DLBCL group, there were significantly higher proportions of patients with older age (≥60 years), International Prognostic Index (IPI) ≥3, Eastern Cooperative Oncology Group (ECOG) scores ≥ 2, bone marrow involvement, positive B-cell lymphoma 2 expression, and positive MYC expression. Survival analysis showed that CD5+ DLBCL had a markedly poorer 2-year progression-free survival than CD5− DLBCL (18.2% vs. 56.2%). Univariate analysis indicated that age ≥60 years, ECOG score ≥ 2, IPI ≥ 3, B symptoms, and no rituximab-based treatment were poor predictive factors for overall survival (OS). Multivariate analysis revealed that B symptoms and no rituximab-based treatment, but not positive CD5 expression, were independent factors for OS.ConclusionsPatients with CD5+ DLBCL had heterogeneous clinical characteristics and poor survival. The development of more targeted and effective therapies is needed.

Subcutaneous rituximab in patients with diffuse large B cell lymphoma and follicular lymphoma: Final results of the non-interventional study MabSCale.

Rituximab has become a standard treatment for non-Hodgkin lymphoma. Clinical studies have demonstrated the efficacy of rituximab in combination with standard chemotherapies in the treatment of follicular lymphoma (FL) and diffuse large B cell lymphoma (DLBCL) patients. This non-interventional study aimed to evaluate the effectiveness and safety of subcutaneous (SC) rituximab in routine clinical practice. Adult patients with previously untreated CD20 positive DLBCL or FL who received rituximab SC and chemotherapy as first-line treatment were observed between 07/2014 and 07/2019 at 99 institutions in Germany. Primary endpoint was the (unconfirmed) complete remission (CR/CRu) rate. Primary outcome was analyzed inferentially; other variables were evaluated descriptively. Overall 583 patients (247 FL; 336 DLBCL) were evaluated. CR/CRu rates were 51.4% (95% CI: 45.2; 57.6) in the FL set and 48.5% (95% CI: 43.2; 53.8) in the DLBCL set. Regarding progression-free survival in the FL group, the probability of being event-free was 94.2% in the first year and 86.2% in the second year. An overall response was achieved in 85.8% (FL) and 85.4% patients (DLBCL). Patient satisfaction at the end of study with the time saving simplification of the SC vs. intravenous route was 98% for FL and 97% for DLBCL. 45.3% of FL and 47.0% of DLBCL patients experienced an adverse event of grade ≥3. Serious adverse events of grade ≥3 occurred in 27.9% FL and 32.4% DLBCL patients, with the highest incidences for leucopenia, anemia, nausea, and fatigue. No new safety signals were detected. The results confirmed the effectiveness and safety of rituximab SC in both the FL and the DLBCL group. Satisfaction of patients and nurses with SC administration was high.

Lkb1 aggravates diffuse large B-cell lymphoma by promoting the function of Treg cells and immune escape

BackgroundRegulatory T cells (Tregs) induce immune responses and may contribute to immune escape in tumors. Accumulation of Tregs in tumors represents a critical barrier to anti-tumor immunity and immunotherapy. However, conflicting results describing the role of Tregs in lymphoma warrant further investigation. The precise features and mechanisms underlying the alteration in Tregs in diffuse large B-cell lymphoma (DLBCL) are not well understood yet. In this study, we analyzed the mechanism underlying the observed alterations in Tregs in DLBCL and examined the effect of Lkb1 expression on the immunosuppressive function of human Tregs.MethodsFlow cytometry and immunofluorescence were used to analyze the proportion of Tregs and effector Tregs in the peripheral blood and lymph nodes of patients with DLBCL and control group. In vitro culture assays were used to analyze the immunosuppressive function of Tregs in the two groups. Transcriptome sequencing was performed to analyze the differentially expressed genes in the two groups, and the transcription level and protein expression of Lkb1 in the two groups were detected using RT-PCR and WES microprotein technology. Lentiviral vectors were constructed to explore the functional changes of Tregs with stable upregulation and downregulation of Lkb1. Finally, a humanized murine lymphoma model was established to study the function of Lkb1 in Tregs in the pathogenesis of DLBCL.ResultsThe number of Tregs was found to be dramatically increased in peripheral blood and tumor tissue in DLBCL patients compared with that in healthy controls, and decreased after treatment. Tregs from DLBCL patients exhibited multiple enhanced functions, including increased inhibition of CD8+cytotoxic T cells (CTL) against tumor cells, enhanced suppression of CD8+CTL secretion of granular enzyme, and suppression of CD8+CTL degranulation. Lkb1 was found to be upregulated in Tregs of DLBCL patients. Furthermore, Lkb1 contributes to Treg immunosuppressive function in DLBCL by regulating the mevalonate pathway. Finally, deletion of Lkb1 in Tregs suppressed tumor growth and promoted anti-tumor immunity in a DLBCL murine model.ConclusionsThese findings confirmed that Lkb1-regulated Tregs are critical for immune escape in DLBCL, which emphasizes that Lkb1 is a potential target for the immunotherapy of DLBCL.

Correlation between Peripheral Blood Intermediate Monocytes Increased and the Disease Progression of Patients with Diffuse Large BCell Lymphoma

The percentage of peripheral blood monocyte subsets of 82 DLBCL patients (including 32 newly diagnosis, 29 remission and 21 relapse) and 30 healthy controls were detected by flow cytometry, and the correlation with the clinical characteristics and its diagnostic value of DLBCL were analyzed. The proportion of intermediate monocytes in patients with newly diagnosed DLBCL group was higher than that in healthy controls (t=5.888, P<0.01). The proportion in relapsed group was higher than those in newly diagnosed DLBCL group（t=2.106，P=0.04） and remission group （t=6.882， P＜0.01）, and the proportion of intermediate monocytes in newly diagnosed DLBCL group was higher than that in Remission group (t=3.969, P<0.01). With the increase of International Prognostic Index (IPI) score, the percentage of intermediate monocytes in patients with DLBCL increased (r=0.37). Furthermore, when the proportion of intermediate monocytes was 10.91% as the cutoff value, the sensitivity and specificity of the whole sample were 90.60% and 9100%, respectively. The disease progression is related to the increased intermediate monocytes, which can be used as a potential diagnostic index for DLBCL.