non-GCB Type Research Articles

Diffuse Large B Cell Lymphoma (DLBCL) Experience in a Majority-Minority Community: The UT Health San Antonio Story

BACKGROUND: Diffuse Large B Cell Lymphoma (DLBCL) is the most common hematologic malignancy in adults. Unfortunately, more than one-third of patients will fail first line therapy, highlighting the need for a better understanding of biology of DLBCL and whether biological variables with clinical significance have the same impact across different ethnicities. Two clinically applicable prognostic models in DLBCL are the cell-of-origin (COO) classifier and the MYC/BCL2 expression pattern. The former classifies DLBCL in germinal center B-cell (GCB) and non-GCB subtypes. Patients diagnosed with GCB-like DLBCLs have significantly better outcome than those with non-GCB-like lymphomas. More recently, IHC-based quantification of MYC/BCL2 in DLBCL biopsies was shown to also impart prognostic value, with tumors positive for both markers, named dual-expresser (DE) DLBCL, harboring a particularly poor outcome. Moreover, DLBCL cases are further characterized into "double hit" (DH) or "triple hit" (TH) based on the rearrangements of MYC and BCL2, MYC and BCL6 or MYC, BCL2 and BCL6, which by far exhibit the poorest outcome. The paucity of data comparing the DLBCL biological variables and outcomes among different ethnicities establishes the basis of our analysis at the only NCI-designated cancer center of south Texas within a Hispanic (HI) predominant patient population. METHODS: This is an observational study where we identified patients with diagnosis of Lymphoma (Hodgkin and Non-Hodgkin) by International Classification of Diseases (ICD) codes. Only patients with DLBCL were retrospectively analyzed; the patients received care at UT Health San Antonio between 2008-2018. Key variables for each patient included age, gender, race/ethnicity, comorbidities, insurance status, stage, treatment received, characterization by COO, IHC markers, molecular rearrangements, outcome at 3 and 5 years and vitality status in 2018. Continuously distributed outcomes were summarized with the mean and standard deviation and categorical outcomes were summarized with frequencies and percentages. The primary endpoint is to compare whether there are differences between HI and non-Hispanics (NH) with respect to biologic variables and hence their outcomes. The study was approved by the local Institutional Review Board. The findings will be available to patients, funders and medical community through traditional publishing and social media. RESULTS: A total of 196 newly diagnosed patients met inclusion criteria, with 115 HI (59%) and 81 NH (41%). HI have a higher percentage (40%) of non-insured, Medicaid or a financial assistance program than NH (28%) . We were able to identify 172 patients with either GCB or non-GCB type. From our HI, 55 (48%) patients were GCB and 46 (40%) were non-GCB while within NH, 47 (58%) patients were GCB and 24 (30%) non-GCB. We gathered IHC data on 147 patients; DE with BCL2/MYC was found in 9 (8%) of HI patients and 14 (17%) of NH patients. There was around the same percentage of unavailable COO and IHC data for both groups, 12% and 25% respectively. Only 11 patients were identified as DH or TH. Among HI, 4 were DH and 1 was TH; while in NH, 5 were DH and 1 was TH (Fig 1A). When looking at survival, as of today 63% of patients are still alive in both groups. Among alive HI, 34 (62%) patients are of GCB origin and 31 (67%) are of non-GCB origin. On the other hand; within the alive NH patients, 34 (72% of GCB positive) are of GCB origin and 11 (46%) are of non-GCB origin (Fig 1B, 1C). Also, DE status apparently had no major difference in survival across ethnicities since 7 (50%) NH remain alive versus 4 (44%) HI (Fig 1D, 1E). Lastly, within HI there are only 2 DH and no TH patients alive, while within NH there are 3 DH patients alive 1 TH patient who is still not 3 years out. CONCLUSION: HI have similar and well-balanced molecular characteristics and outcomes compared to NH. As opposed to what we thought, the COO classifier does not seem to predict differences in prognosis in HI as they appear to have equal outcome irrespective of GCB vs non-GCB origin. The prognosis in HI with overexpression of MYC/BCL2 is bad and even worse if DH or TH as historically seen in NH. To our knowledge this is the first retrospective review that includes the largest Hispanic cohort in North America. Figure 1 Disclosures Hernandez: ASH MRHAP: Research Funding.

PF530 CORRELATION OF CD10 EXPRESSION DETERMINED BY FLOW CYTOMETRIC IMMUNOPHENOTYPING AND CELL-OF-ORIGIN DETERMINATION BY GENE EXPRESSION PROFILING IN DIFFUSE LARGE B-CELL LYMPHOMA

Background: Diffuse large B-cell lymphomas (DLBCL) represent a clinically heterogeneous group that is classified together based on similarities in morphology and immunophenotype. However, it was discovered that gene expression profiling (GEP) could classify DLBCL into distinct molecular subgroups based on cell-of-origin (COO), including germinal center B-cell type (GCB), activated B-cell type (ABC), and unclassified (UNC) type. COO assignment of DLBCL has important biological and prognostic significance, as well as potential therapeutic implications, with the ongoing development of selective agents for treatment of specific DLBCL subtypes. Indeed, the current WHO Classification for lymphoma requires COO determination for all cases of DLBCL at diagnosis using either GEP or immunohistochemistry methods. Flow cytometric immunophenotyping is also commonly used in the diagnostic workup of DLBCL, and we were interested to determine how well CD10 expression determined by flow cytometry correlated with GCB type DLBCL using a clinical Lymph2Cx GEP COO assay. Here, we summarize the results of 74 clinical DLBCL cases analyzed by both flow cytometric immunophenotyping and Lymph2Cx COO testing. Aims: We wished to determine the predictive value of CD10 expression determined by flow cytometric immunophenotyping for the presence or absence of GCB type DLBCL established by GEP. Methods: 74 clinical DLBCL cases were subjected to Lymph2Cx GEP COO analysis performed on the NanoString nCounter® Digital Analyzer and flow cytometric immunophenotyping with CD10 expression using a BD FACSCanto II flow cytometer. Hans algorithm immunohistochemical stains (CD10, BCL6, and MUM1) were also performed on 65 of the 74 cases. Results: Lymph2Cx COO testing detected 40 GCB, 25 ABC, and 9 UNC cases. 36 cases (49%) were positive for CD10 expression by flow cytometric immunophenotyping, and 38 cases were negative for CD10 expression (51%). 32 of the 36 CD10-positive cases were determined to GCB type by Lymph2Cx testing. 30 of the 38 CD10-negative cases were determined to be either ABC or UNC type by Lymph2Cx analysis. The predictive value of CD10 expression by flow cytometry was 89% for GCB type DLBCL by GEP COO analysis. In CD10-negative cases, the predictive value for the presence of ABC or UNC type DLBCL was 79%. Interestingly, CD10-positive cases by flow cytometry also correlated with GCB type DLBCL determined by Hans algorithm immunostains in 34 of 35 cases (97%), but only 31 of these cases (89%) were true GCB-DLBCL by GEP analysis. However, lack of CD10 expression by flow cytometry correlated with non-GCB type DLBLC by the Hans algorithm in only 21 of 30 cases (70%), with nine cases exhibiting a GCB type immunohistochemical profile. Summary/Conclusion: In summary, we have demonstrated that the presence of CD10 expression in DLBCL determined by flow cytometric immunophenotyping shows a very strong correlation with GCB type by Lymph2Cx GEP COO testing. CD10 expression by flow cytometric immunophenotyping also correlates well with GCB designation by the Hans algorithm immunohistochemical staining. Lack of CD10 expression by flow cytometric immunophenotyping also showed good correlation with ABC or UNC type DLBCL determined by GEP. These findings suggest that when clinical samples are limiting and only limited immunophenotyping data is available, the presence or absence of CD10 expression determined by flow cytometric immunophenotyping can be helpful for predicting COO in DLBCL.

MDS AS A CAUSE FOR PROLONGED HEMATOLOGIC TOXICITY AFTER TREATMENT WITH CD19 TARGETED CAR‐T CELL THERAPY IN PATIENTS WITH RELAPSED REFRACTORY LYMPHOMA

Introduction: The advent of adoptive cellular therapy utilizing autologous T cells expressing chimeric antigen receptors (CAR-T) against CD19 has revolutionized relapsed and refractory B cell lymphoma treatment. CAR-T cell therapy, however, is not without toxicity. Other than cytokine release syndrome and neurotoxicity, hematologic toxicity occurs commonly as well. Grade 3 and above prolonged neutropenia and thrombocytopenia (longer than 4 weeks since T cell infusion) was seen in 15 to 40% and 18 to 40% of patients treated with CD19 CAR T cells in clinical trials respectively. The exact mechanism underlying the pathogenesis of prolonged hematologic toxicity in CAR-T cell therapy is not well understood. Interestingly, we identified four cases of prolonged cytopenias after CAR-T cell therapy due to the presence of myelodysplastic syndrome in patients receiving this treatment for relapsed refractory B cell lymphoma. Methods: We performed a retrospective review of four patients with relapsed refractory aggressive B cell lymphoma treated at our institution with CD19 directed autologous CAR-T cells. Results: Median age was 74 (range: 57-76). Two patients had GCB type, 2 had non-GCB type DLBCL. The median number of lines of therapy prior to CAR-T cell therapy was 5. All four patients had prior autologous stem cell transplantation. Median time to MDS diagnosis from CAR T infusion was 3 months (range: 2 to 3). One patient developed MDS after additional chemotherapy was given for relapsed disease. One patient had evidence of a dysplastic clone prior to CAR-T cell therapy. The remaining two patients did not have evidence of MDS until immediately after CAR-T cell therapy. Two patients developed severe CRS requiring tocilizumab and corticosteroids. One patient showed partial response to CAR-T cell therapy, while three others achieved CR, two of which have since relapsed. Discussion: We identified four patients with prolonged hematologic toxicity after CAR-T cell therapy due to MDS. These patients have multiple risk factors for the development of MDS including advanced age, and heavy pretreatment. In fact, one patient had evidence of MDS prior to CAR-T cell therapy, underscoring the potential benefit of providing CAR-T cell therapy earlier in the course of treatment. In addition, our findings suggest that a bone marrow study should be performed in patients with prolonged cytopenias post CAR T cell therapy. It is not clear if there is any direct contribution of CAR T cells and the resulting immune system disturbance to the development of MDS. Long term follow up of all patients receiving CAR T cell therapy is therefore crucial to assessment true incidence of MDS and establish association. Keywords: B-cell lymphoma; CD19. Disclosures: Herrera, A: Consultant Advisory Role: Gilead. Siddiqi, T: Consultant Advisory Role: Juno Therapeutics. Budde, E: Consultant Advisory Role: Gilead. 480 – WITHDRAWN

Correlation between the immunophenotypes, treatment strategies and prognosis of gastrointestinal diffuse large B-cell lymphoma

Objective To explore the differences in the prognosis of patients with different immunophenotypes gastrointestinal diffuse large B-cell lymphoma (GI-DLBCL) who received different treatment strategies. Methods From March 2006 to January 2016, at Nanfang Hospital, Southern Medical University in Guangzhou, the clinical data of 99 patients with pathologically confirmed GI-DLBCL were retrospectively analyzed. According to treatment strategies, patients were divided into chemotherapy alone group and combination of surgery and chemotherapy group. According to immunophenotypes, patients were divided into germinal center B-cell-like (GCB) type and non-GCB type. The two-year progression-free survival (PFS) rate and overall survival (OS) rate were evaluated. Kaplan-Meier analysis, log-rank test and Cox regression were performed for statistical analysis. Results Among the 99 patients with GI-DLBCL, 51 patients were treated with chemotherapy alone, and 48 patients were treated with combination of surgery and chemotherapy. Forty-one cases were GCB phenotype and 40 cases were non-GCB phenotype. The median follow-up time was 25 months. The two-year PFS and OS rates were 70.9% and 89.5%, respectively. The two-year PFS and OS rates of chemotherapy alone group were 63.6% and 85.0%, respectively; both were lower than those of combination of surgery and chemotherapy group (79.4% and 94.7%), and the differences were statistically significant (χ2=4.232, P=0.040 and χ2=4.260, P=0.039). The two-year PFS and OS rates of GCB group were 68.8% and 93.9%, respectively. And the two-year PFS and OS rates of non-GCB group were 73.2% and 85.6%, respectively. There were no statistically significant differences between these two groups (both P>0.05). Among 41 patients with GCB type, 25 were treated with combination of surgery and chemotherapy and 16 were treated with chemotherapy alone. The two-year PFS rate of patients treated with combination of surgery and chemotherapy (83.1%) was higher than that of patients treated with chemotherapy alone (49.2%), and the difference was statistically significant (χ2=5.627, P=0.018). The results of multivariate analysis indicated that treatment strategy was not an independent prognostic factor for all the enrolled patients and in patients with GCB type (all P>0.05). Conclusions Immunophenotypes may lack evaluation value of prognosis in patients with GI-DLBCL. Although among all the enrolled patients and patients with GCB type, the prognosis of patients treated with combination of surgery and chemotherapy is better than that of patients treated with chemotherapy alone, treatment strategy is not an independent prognostic factor. Multi-factors should be evaluated before selection of treatment strategy. Key words: Gastrointestinal tract; Lymphoma, large B-cell, diffuse; Surgery; Chemotherapy; Prognosis

Genomics of Limited-Stage Diffuse Large B-Cell Lymphoma Developing Late Relapse: Analysis of Gene Alterations in Paired Primary and Late Relapsed Tumors By Target Sequencing

Genomics of Limited-Stage Diffuse Large B-Cell Lymphoma Developing Late Relapse: Analysis of Gene Alterations in Paired Primary and Late Relapsed Tumors By Target Sequencing

Cell-of-Origin Identification and Prognostic Correlation in HIV-Associated Diffuse Large B-Cell Lymphomas: Results of an Italian Multicentric Study

Introduction: Diffuse Large B-cell lymphoma (DLBCL) is an heterogeneous disease in terms of biology, clinical presentation and prognosis. According to the putative Cell-Of-Origin (COO), DLBCL of the general HIV negative population can be classified into three molecular subtypes by gene expression profiling (GEP): the germinal center B-cell type (GCB), the activated B-cell type (ABC) and the unclassifiable type (UC). Immuno-histochemistry (IHC) models routinely surrogate GEP in the COO assignment, identifying GCB and non-GCB type. Hans algorithm is one of the most widely used IHC models and its concordance with GEP varies from 70 to 90% (Meyer PN et al, JCO 2011). NanoString (NS) 20-genes assay has emerged as a feasible technique for DLBCL molecular subgroups identification and shows a high concordance with GEP (Scott DW et al, Blood 2014). DLBCL classification according to COO categories has shown prognostic impact in the HIV negative population, since clinical outcome of non-GCB type is inferior if compared to GCB type in patients (pts) receiving CHOP or Rituximab-CHOP. DLBCL in HIV positive population have shown different biological features. COO subtypes distribution and its prognostic impact has not been extensively studied in HIV setting and still needs to be defined.Aim of the study: To evaluate the proportion of COO subtypes in HIV-associated DLBCL according to IHC and NS assay, the concordance of these two different diagnostic methods, and the prognostic impact of COO assignment; to analyze BCL-2 and MYC protein expression and their correlation with outcome; to determine the prognostic value of baseline clinical characteristics, such as stage, LDH value and CD4+ lymphocyte count.Methods: We retrospectively evaluated 66 cases of HIV positive pts with newly diagnosed DLBCL from 2000 to 2016, in 5 Italian Institutions. Histological samples were centrally reviewed by a panel of expert Pathologists for DLBCL diagnosis confirmation, BCL-2 and MYC protein expression and COO assignment according to Hans algorithm (Hans CP et al, Blood 2004). The cut off considered for BCL-2 and MYC overexpression was 70% and 40% respectively. NS Lymph2Cx assay for COO was performed in all pts. Clinical data were gathered from pts medical records.Results: Pts were mostly male (73%) and median age at DLBCL onset was 45 years (range: 28-83). 80% of pts had advanced stage lymphoma and 34% showed a high-intermediate or high IPI score. HIV first detection was concomitant to DLBCL diagnosis in 30% of pts. At DLBCL diagnosis median CD4+ cell count was 188/microL (range: 8-1172) and 34/66 pts (52%) had detectable HIV viral load. COO assigned by IHC was GCB in 31/66 pts (47%) and non-GCB in 35/66 (53%); COO allocation by NS assay was 57% GCB (n=34/60 pts) and 43% non-GCB (ABC n=11, 18%; UC n=15, 25%). IHC algorithm and NS assay concordantly assigned COO subtypes in 80% of pts with a Cohen's kappa=0,604 (p<0.0005). Twenty pts (30%) had MYC and 23 pts (33%) had BCL-2 overexpression, respectively. An anthracycline-containing first-line chemotherapy was delivered to 97% of pts, with the addition of Rituximab in 86% of cases. Two pts (3%) could receive only palliative treatment. Two-years progression-free survival (PFS) and overall survival (OS) of the entire series were 61% and 56%, respectively, after a median follow-up of 50 months (range 4-184). No difference was found in OS and PFS according to COO subtypes, either determined by IHC or NS. Notably, bcl-2 overexpression had a negative impact on OS (2-years OS 44% vs 70%, p=0.018, HR: 2,34) and PFS (2-years PFS 45% vs 62%, p= 0,12, HR: 1,71), while MYC expression had no significant correlation with survival. A baseline CD4+ count > 200/microL has a protective role both for progression and death (2-years PFS: 62% vs 42%, p=0.02, HR: 0.41; 2-years OS: 72% vs 42%, p=0.002, HR: 0.30). Stage IV disease and elevated baseline LDH were associated with inferior PFS (p=0.0158 and p=0.02) and OS (p=0.0076 and p=0.034).Conclusions: This analysis of 66 HIV-associated DLBCL confirmed that GCB is the more frequent subtype identified by NS, as previously reported (Baptista MJ et al, Hemat Oncol 2017). We found an high concordance between Hans algorithm and NS in the COO subtypes allocation. However, COO classification showed no impact on pts outcome. FISH assays for detection of MYC, BCL-2 and BCL-6 rearrangements are ongoing in the entire study population. DisclosuresRusconi:Celgene: Research Funding.

Diffuse Large B Cell Lymphoma in an Adolescent: A Challenging Case with Intracerebral and Retinal Involvement

Diffuse large B-cell lymphoma (DLBCL) is a rare lymphoma type in pediatric patients with usually good prognosis. We present the difficult diagnostic procedure and the complicated course in an adolescent patient with DLBCL.Case report: The 16-year-old female patient presented with a history of fever episodes during the past 3 weeks, hepatosplenomegaly, leukopenia, anemia, hypertriglyceridemia, massive hyperferritinemia and elevation of lactate dehydrogenase. Leukemia was excluded by bone marrow puncture. Initially, no significant lymph node enlargement was detected by imaging of the mediastinum and the abdomen but a pathological signal of the bone marrow could be observed in magnetic resonance imaging (MRI). Hemophagocytic lymphohistiocytosis (HLH) was diagnosed and the patient was treated with prednisone 0.5 mg/kg per day. The overall condition and the laboratory values improved but deteriorated on cessation of the steroid treatment. The result of the bone marrow biopsy then revealed an infiltration of a malignant B-cell lymphoma. Diagnosis of a DLBCL, Non-GCB-type with a BCL6-break was subsequently made in a biopsy of a lymphnode which had shown increased uptake in PET scan. Treatment according to the current NHL-BFM registry including administration of Rituximab was started. The course of disease was complicated by HLH symptoms with persistent fever and pancytopenia, renal failure, severe Methotrexate excretion failure requiring Carboxypeptidase treatment and Herpes virus encephalitis. After recovery progression of DLBCL was found and the treatment was switched to DA-EPOCH regimen and repeated systemic Rituximab administrations. Five months after diagnosis the girl suffered from visual impairment and convulsions. Ophthalmoscopy revealed retinal lesions and intracerebral lesions were detected by MRI but no abnormalities of cerebrospinal fluid (CSF) were found. Biopsy of a brain lesion confirmed cerebral involvement of DLBCL. Intensified treatment included high- dose steroids, cranio-spinal irradiation, high-dose Cytarabin and intrathecal Rituximab. Response of the cerebral lesions but no improvement of the visual impairment was found. Salvage treatment with Gemcitabin, Cisplatin and Rituximab was administered because of persistent nodal and extranodal lymphoma manifestations. The patient finally died due to refractory disease and severe bone marrow failure.Conclusion: This case of DLBCL in an adolescent girl is extraordinary in many ways. DLBCL, especially the non-GCB type, is a rare disorder in pediatric patients. Predominant bone-marrow involvement has been described in only few older patients with non-GCB-type DLBCL and mostly aggressive clinical course. HLH as a presenting feature is rather observed in T- or NK-cell lymphoma than in DLBCL. Finally, central nervous system (CNS) relapse occurs in only about 6% of adults with DLBCL regardless of CNS prophylaxis. Retinal and intraparenchymal CNS involvement without abnormalities in the CSF has been typically described for primary CNS lymphoma in adults. All these conditions have contributed to the adverse outcome in our patient. DisclosuresNo relevant conflicts of interest to declare.

Impact of MYD88L265P mutation Status on Histological Transformation of Waldenstrom Macroglobulinemia

Impact of MYD88L265P mutation Status on Histological Transformation of Waldenstrom Macroglobulinemia

High Level of Pre-Treatment C-Reactive Protein-to-Albumin Ratio Predicts Inferior Prognosis in Diffuse Large B Cell Lymphoma

IntroductionDiffuse large B cell lymphoma (DLBCL) is the most common highly aggressive non-Hodgkin lymphoma (NHL) worldwide. The International Prognostic Index (IPI) has been established as a useful prognostic marker, and there have been some proposed markers which could reflect tumor microenvironment including neutrophil, lymphocyte, platelet, serum globulin, ferritin and serum free light chain. C-reactive protein (CRP) is one of the commonly used inflammatory markers, and its clinical relevance has been suggested recently in various malignancies. Serum albumin is a representative marker for nutritional status, and previous studies have presented that hypoalbuminemia might be an indicator of cancer-related inflammation as well. In this point of view, C-reactive-to-albumin ratio (CAR) has been suggested as one of easily-accessible parameters which could be a robust prognostic marker in diverse malignancies such as lung cancer, gastric cancer and colorectal cancer. However, its clinical value has not been assessed in hematologic malignancies. In this study, we evaluated the prognostic effect of CAR in DLBCL.MethodsThis retrospective study included 186 patients who were histologically diagnosed with DLBCL and treated with R-CHOP (Rituximab, Cyclophosphamide, Doxorubicin, Vincristine and Prednisone) between 2006 and 2018 at National Cancer Center, Korea. One hundred forty one cases were identified whose baseline laboratory values including CRP and albumin were available, and then the medical records were reviewed. To define the appropriate cutoff value of CAR in patients with DLBCL, cutoff finder method was applied which had been suggested by Budczies et al., and the most discriminative point was designated by the value of 0.158. Clinical characteristics and outcomes including response rate, overall survival (OS) and progression-free survival (PFS) were investigated between high and low CAR group. Additionally, the clinical value of CAR was compared to the components of IPI for DLBCL as well.ResultsOf all patients, 73 (51.8%) were classified as high CAR group. Male was 42 (57.5%) in high CAR group and 37 (54.4%) in low CAR group. In terms of IPI, 21 (28.8%) were classified into high IPI (score of 4 or 5) in high CAR group - 6 (8.8%) in low CAR group in comparison. Hans criteria was applied to discriminate germinal center B-cell (GCB) subtype to non-GCB subtype by immunohistochemistry and after 12 patients removed due to missing GCB status, 56 (84.8%) in high CAR group was sorted to non-GCB type - 46 (73.0%) in low CAR group to be compared. The high CAR group showed significantly worse complete response (CR) rates to induction R-CHOP therapy (64.4% vs. 92.6%; p<0.001). Median cycles of induction chemotherapy was 6 in all patients and there was no significant difference between both groups (p=0.824). With a median follow-up of 32.5 months, the high CAR group revealed significantly worse 5-year OS (65.0 vs. 93.5%; p<0.0001) and 5-year PFS (53.5 vs. 80.7%; p<0.0001) (Figure 1). In univariable Cox analysis, high CAR was a statistically significant prognostic factor for both 5-year OS (HR 8.04, 95% CI 2.384-27.139; p=0.001) and 5-year PFS (HR 4.44, 95% CI 2.109-9.341; p<0.001). Along with CAR, all components of IPI was statistically significant for both OS and PFS, except age at diagnosis (HR 1.95, 95% CI 0.908-4.207, p=0.087) for 5-year OS. In multivariable analyses with adjustment for age (>60), stage (III, IV), lactate dehydrogenase (LDH) (>upper normal limit), Eastern Cooperative Oncology Group (ECOG) performance status (>1) and the number of extranodal involvement (>1), high CAR showed statistically significant results for both 5-year OS (HR 4.71, 95% CI 1.175-18.892; p=0.029) and 5-year PFS (HR 2.66, 95% CI 1.122-6.289; p=0.026) (Table 1).ConclusionsIn conclusion, CAR might play an additional role to IPI in prognostication of patients with DLBCL considering the fact that it is simple, objective and easy to obtain. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Primary Diffuse Large B Cell Lymphoma of Thyroid Gland; Germinal Center and Non-germinal Center B Types: Experience of a Single Center

Diffuse large B cell lymphoma (DLBCL) is the most common histological subtype accounting for approximately 70% of primary thyroid lymphoma (PTL) cases. Its prognosis depends on the histology, spread, and the stage of the disease. DLBCL usually displays a better prognosis, but it shows heterogeneity according to the histological subtype. Germinal center B cell (GCB) subtype has a more favorable prognosis than the non-germinal center (non-GCB) subtype. Most of these patients required combined modality management. This study included eight patients with stage I/IIE thyroid DLBCL. Analysis of clinicopathologic features of all PTL patients diagnosed at Cerrahpasa Faculty of Medicine. Pathology reports and slides of patients diagnosed between years of 2000 and 2013 were evaluated. Four of them were GCB and the other four cases were non-GCB type. All patients underwent thyroidectomy. Three of them were treated with chemotherapy and the three patients were treated with chemotherapy followed by radiotherapy. Five patients are presently alive. Due to the fact that PTLs are rare types of tumors, only small case series were published in that field. Increasing the number of cases in the literature for thyroid DLBCL, its histopathologic features will provide a better understanding.

Immunohistochemical distinction of ABC and GCB in extranodal DLBCL is not reflected in mutation patterns

Gene expression profiling (GEP) separated diffuse large B-cell lymphoma (DLBCL) in two different entities, i.e. activated B cell-like (ABC) and germinal center B cell-like (GCB) lymphomas with ABC lymphomas demonstrating a less favorable outcome. NF-kB pathway activating mutations in MYD88, CD79A/B and CARD11 are predominantly found in ABC type lymphomas. Targeted therapies affecting NF-kB pathways have shown therapeutic potential in this subtype. Immunohistochemistry algorithms have been developed as a tool for distinguishing these entities in routine clinical diagnostics. To test whether this immunohistochemistry classifier would detect the biological differences between the entities 147 DLBCLs were subtyped into ABC and GCB using the Visco-Young algorithm. Mutation analysis demonstrated mutations in MYD88 or CD79 A/B in 21% (10/47) of non-GCB type but only in 3% (1/31) of GCB lymphomas (p = 0.012) in nodal lymphomas. In primary extra nodal lymphomas, however, 17.5% (4/23) of GCB type and 37.5% (15/40) of non-GCB lymphomas carried mutations in MYD88 and CD79 A/B. While the Visco-Young algorithm was sufficient to detect biological differences (i.e. mutation patterns) in nodal DLBCL it did not distinguish GCB and non-GCB type lymphomas of primary extranodal sites. Here, the morphological sites of the lymphomas seem to be more important for their molecular pattern than their immunohistochemical status.

Expression and Clinical Prognostical Significance of RIP2 in Diffuse Large B Cell Lymphoma

To investigate the expression and prognosis significance of receptor interacting protein 2 (RIP2) in diffuse large B-cell lymphoma (DLBCL). The expression of RIP2 in DLBCL GCB and non-GCB type was detected by immunohistochemistry, at same time the expressions of BCL-2 and C-MYC were detected. Then, the role of RIP2 in development of DLBCL was analyzed by related clinical and pathological parameters. The expression of RIP2 was related with middle-high risk group by IPI score, the An Arbor stage III+IV and intranodal lesions, and the differences were statistically significant (P<0.05). Besides, the single factor survival analysis suggested that GCB-type DLBCL showed a higher survival rate than that in non-GCB type(P<0.05). Patients with RIP2+ showed a lower survival rate as compared with patients with PIP2- (P<0.05), among which the patients receiving R-CHOP had a higher survival rate than that of those receiving CHOP (P<0.01). The expression of RIP2 in DLBCL cell lines was higher than that in peripheral mononuclear cells of normal subjects (P<0.01) and expressed differently in DLBCL of GCB and non-GCB type (P<0.01). The expression of RIP2 may relate with the poor prognosis and specific subtype of DLBCL.

Prevalence and Prognostic Significance of CD5 Immunohistochemistry in De-Novo DLBCL in a Western Population Cohort

Abstract Background: Various subsets of Diffuse Large B-cell lymphoma (DLBCL) are distinguished based on molecular and Immunohistochemical features. CD5 expressing DLBCL (CD5+ DLBCL) is increasingly recognized as a subtype of DLBCL with an aggressive disease course in small elderly Asian population cohorts comprising 5-10% of de-novo DLBCL. We attempt to describe its prevalence and prognostic implications in concert with IHC expression profiles of biomarkers Methods: A retrospective review of elderly patients (65yr-92yr) with de-novo Diffuse Large B-Cell lymphoma (DLBCL) uniformly treated with RCHOP was conducted between Jan 98 and Dec 2016. Patient demographics, clinical parameters and IHC markers were collected. Analysis using chi square, or Fisher exact where appropriate, was performed. Kaplan-Meier analysis and univariate and multivariate Cox regression analysis identified associations with overall survival. The protocol, and subsequent modifications, were reviewed and approved by the IRB at the Texas Tech University Health Sciences Center Results: One hundred fourteen de-novo DLBCL with known CD5 IHC status, Ki67, c-MYC, BCL2, BCL6, MUM1, CD10 marker status and Han's classifiers were evaluated. Median age of the CD5 negative cohort was 75yrs ±12.8 vs 80yrs±9.3 in the CD5+ subgroup (p=0.51). Median OS 6.7 years (SD 62.3mo; Range 1-256). CD5+ expression on IHC was seen in 23/110 (21%) patients. IHC marker status showed c-myc (12%), BCL2(42%) and BCL6(39%) respectively. Compared with matched group of patients with CD5 negative DLBCL, CD5+ DLBCL were more likely to be non-GCB type (14/23 vs 9/87; p 2 (14/23 vs 23/81; p=0.009) and Stage >III (p=0.004). Three patients with CNS involvement all had CD5 expression. Kaplan-Meier analysis showed that Han's non-GCB classification (p 70% (HR=0.33, P=0.017), BCL6 (HR=0.47, P=0.019) and CD10 (HR=0.2, P=0.003) were associated with worse outcomes. CD5+ expression was not significantly associated with OS (HR=0.76; -1.4 to 0.8; p=0.64) in multivariate model, however was significant in a univariate analysis (P Conclusion: CD5+ DLBCL subset is more common in this elderly western cohort than historically seen in the Asian population with a signal toward Hans non-GCB, higher R-IPI, higher Stage and perhaps higher risk of CNS involvement. More data is needed to evaluate if CD5+ de-novo DLBCL has any correlation with other antiapoptotic or proliferative markers like BCL2,6 and c-MYC and larger patient subset and prospective studies are needed to corroborate these findings Disclosures No relevant conflicts of interest to declare.

Difference of Clinical Parameters between GCB and Non-GCB Subtype DLBCL

Abstract Background Diffuse large B cell lymphoma (DLBCL) is divided into Germinal center B-cell like (GCB) subtype and non-GCB by the Hans algorithm and non-GCB is known to be associated with unfavorable prognosis. Therefore, we tried to compare the difference of clinical parameters between GCB and non-GCB DLBCL subtypes. Methods We retrospectively analyzed DLBCL patients in the Asan Medical Center's lymphoma database from January 2001 to December 2014. We selected 796 patients who received first line R-CHOP chemotherapy and whose cell-of-origin can be classified by the Hans algorithm. Results Among the 796 patients, 261 (32.8%) were in the GCB subtype and 535 (67.2%) were in the non-GCB group by the Hans algorithm. ECOG PS≥2, increased β2M (β2-microglobulin), increased LDH, Ann Arbor stage 3-4, high NCCN IPI and GELTAMO IPI stage were associated with non-GCB type. There was borderline significant difference in disease-free survival (DFS) for patients in the GCB and non-GCB cohorts (p=.07). Conclusions Non-GCB subtype DLBCL had relatively unfavorable prognostic factors than GCB DLBCL. Such as ECOG PS≥2, increased β2M (β2-microglobulin), increased LDH, Ann Arbor stage 3-4, high NCCN IPI and GELTAMO IPI stage were more distributed in non-GCB DLBCL. Disclosures No relevant conflicts of interest to declare.

Advances in Diagnosis and Management of Diffuse Large B-cell Lymphoma.

The management of diffuse large B-cell lymphoma (DLBCL) has been gradually evolving since the discovery of its 2 major forms, the germinal center B-like (GCB) and activated B-cell (ABC) types. Although the reference standard for the identification of these cell types is considered gene expression profiling (GEP), currently the only method commercially available is immunohistochemistry (IHC). The application of various IHC-based algorithms and their correlation with GEP and clinical outcome are discussed. Because of the adverse prognostic implications of the non-GCB type and its potential effects on treatment selection, the recently revised World Health Organization classification has included these biologic cell types. The management of double hit lymphomas, which almost exclusively fall under the GCB category, is discussed, together with the double expresser phenotype, which is usually grouped under the non-GCB type. The role of lenalidomide and ibrutinib in the management of the non-GCB type is examined. We also discuss the front-line management of primary mediastinal large cell lymphoma using the EPOCH (etoposide, prednisolone, Oncovin [vincristine], cyclophosphamide, hydroxydaunorubicin [doxorubicin]) regimen and examine new salvage data on immune checkpoint inhibitors for this clinical subtype. The prognosis, clinical features, and management of de novo CD5+ DLBCL are discussed, and newer and promising developments in the management of primary central nervous system lymphomas are presented in detail. The most popular salvage regimens and the application of high-dose chemotherapy with stem cell transplantation are assessed in detail. Finally, data on new treatment tactics such as CART (chimeric antigen receptor T-cell) cells and promising new drugs, including blinatumomab and venetoclax, are presented.

Immunohistochemical Expression of CD10, BCL6 and MUM1 in Differentiating Diffuse Large B Cell Lymphoma Subtypes.

To determine the immunohistochemical expression of CD10, BCL6 and MUM1 in differentiating diffuse large B cell lymphoma subtypes. Descriptive, cross-sectional study. Department of Histopathology, Armed Forces Institute of Pathology, Rawalpindi, from November 2014 to May 2015. Newly diagnosed cases of DLBCLon H&E stain as well as IHC markers, according to WHO blue book 2008, were included in the study. Patients' gender, age and site of lymphoma were noted. DLBCLsubtypes (GCB and activated type or non-GCB) were assessed based on IHC expression of CD10, BCL6 and MUM1 and the results were recorded. The data were analyzed by using computer software program SPSS version 20. Descriptive statistics, frequencies and percentages were calculated. Out of 96 patients, 79 (82%) were male and 17 (18%) were female. Mean age was 54.66 ±16.73 years. Thirty-six (37.5%) cases showed positivity for CD10 and BCL6 both (GCB type), whereas MUM1 was positive in 60 (62.5%) cases (non-GCB type or activated type). Asignificant statistical association was seen between expression of IHC markers (CD10, BCL6 and MUM1) and DLBCLsubtypes (GCB and non-GCB type, p<0.001). In Pakistani population, the frequency of non-GCB type expressing MUM1 is 62.5%, which is quite high as compared to western countries. It needs to be further explored, because it represents high-risk subsets in which alternative strategies for diagnosis and management should be planned.

Clinicopathologic analysis of ten patients with gonadal diffuse large B-cell lymphoma

Objective To investigate the clinicopathologic characteristics, immunophenotype, differential diagnosis, and prognostic factors of gonadal diffuse large B-cell lymphoma (DLBCL). Methods The clinicopathologic data of 10 patients with gonadal DLBCL including morphology and immunohistochemistry were analyzed retrospectively, then the literature was reviewed. Results Among 10 patients, 9 cases were testicular DLBCL, and the median age was 67 years old (40-85 years old); 1 case was ovary DLBCL, and the age was 46 years old. Tumor cells were large to medium-sized under the optical microscope, which were characterized as diffuse infiltration around the duct and the remaining convoluted tubule of testis neoplasms. Most DLBCL (70%, 7/10) immunophenotype analysis showed non-germinal center B-cell (non-GCB) type. Patients were followed up from 6 to 103 months, and 2 patients lost follow-up. The survival number of patients in 1-year, 3-year, and 5-year was 4, 2, 2 respectively. Conclusions Primary gonadal DLBCL is an uncommon extranodal lymphoma, which mainly belongs to non-GCB type with poor prognosis. Comprehensive treatments usually take operation and chemotherapy, and the prognosis should be evaluated by multiple factors. Key words: Lymphoma, large B-cell, diffuse; Neoplasms, gonadal tissue; Testis; Ovary; Clinical characteristic; Pathological subtype; Prognosis

EZH2 Upregulation Is Associated with Unfavorable Prognosis in Diffuse Large B-Cell Lymphoma through Potential RUNX3 Downregulation

Introduction: The runt-related transcription factor 3 (RUNX3) is a downstream effector of the transforming growth factor-β (TGF-β) signaling pathway, and has critical roles in apoptosis, angiogenesis, cell migration and invasion. Putative tumor suppressor activity of RUNX3 has been presented extensively in the literature, particularly in solid epithelial tumors and recently in lymphoma with loss of expression favoring tumorigenesis and/or prognosis, but its role in diffuse large B-cell lymphoma (DLBCL) has not been studied. Enhancer of zeste homolog-2 (EZH2), a histone methyltransferase, has been shown to mediate silencing of RUNX3. RUNX3 downregulation due to EZH2 upregulation has been shown in various solid tumors. In the present study, we investigated the EZH2 and RUNX3 RNA expression status in DLBCL and its impact on clinical outcome.Methods: A retrospective chart review was performed and 169 cases of DLBCL treated with chemoimmunotherapy between 2003 and 2013 were included. Immunodeficiency- or EBV-associated and MYC+ LBCL were excluded. Archived formalin-fixed-paraffin-embedded tissue samples were retrieved and RNA was extracted using commercially available kits. We correlated the RNA expression levels for EZH2 and RUNX3 in various sites using quantitative real-time PCR (Taqman assay) and custom designed primers for each gene. Control samples included three benign lymph nodes free of a neoplastic process.Results: We identified 66 cases of DLBCL, including25 nodal DLBCL and 41 extranodal DLBCL, with sufficient RNA extracted. Extranodal locations included testis (n=12), orbit (n=6), primary central nervous system (n=5), bone (n=3), breast (n=2) and viscera (n=13). The median age was 64 years (range 29- 81 years) with a female to male ratio of 0.4 (F=20 and M=46). Median overall survival (OS) was 28 months (1-156 months). Immunophenotypic subtype based on cell-of-origin using Hans algorithm was available in 63 cases; 34 cases were germinal center B-cell (GCB) type while 29 were non-GCB type. Treatment data was available in 63 cases and all patients received R-CHOP as initial therapy except three patients who died shortly after diagnosis. Forty-four cases showed higher expression of EZH2 and RUNX3 when compared to normal lymph nodes (p < 0.05). Nineteen out of 44 cases showed increased EZH2 and decreased RUNX3 expression (Group 1) while EZH2 expression was lower than RUNX3 in the remaining cases (Group 2). The remaining 22 DLBCL cases did not show significant correlation for expression (Group 3). Overall survival was significantly low in Group 1 compared to Group 2 and Group 3 (p =0.030 and p=0.026, respectively). There was no difference for OS between Groups 2 and 3 (p>0.05) (Figure 1).Conclusions: Our results showed that decreased RUNX3 RNA expression is associated with EZH2 overexpression and poses an adverse prognostic factor in DLBCL. Larger studies are needed to establish the prognostic and therapeutic utility of EZH2 and/or RUNX3. [Display omitted] DisclosuresMehta:Pharmacyclics: Research Funding; Merck: Research Funding; Incyte: Research Funding; Medimmune: Research Funding; Roche Genentech: Research Funding; Bristol Myers Squibb: Research Funding; Seattle Genetics: Membership on an entity's Board of Directors or advisory committees. Forero:University of Alabama at Birmingham: Research Funding. Costa:Sanofi: Honoraria, Research Funding.

CD30 expression and prognostic significance in R-EPOCH–treated patients with diffuse large B-cell lymphoma

A few recent studies investigated the prognostic impact of CD30 expression in diffuse large B-cell lymphoma (DLBCL) patients treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone. No study has evaluated the significance of CD30 expression in DLBCL patients treated with rituximab, etoposide, prednisone, vincristine, cyclophosphamide, and doxorubicin (R-EPOCH). In a group of 97 patients with DLBCL and high-risk features who received R-EPOCH induction therapy, we studied CD30 expression by immunohistochemistry using different cutoff values (>0% and ≥20% of lymphoma cells, respectively) and correlated with prognosis. CD30 expression was detected in 24 (25%) cases using a cutoff greater than 0% and in 12 (12%) cases using a cutoff of 20% or greater. The clinicopathological features were similar between CD30+ and CD30-negative groups. A major difference was that MYC rearrangement was infrequent in the CD30+ group: 2 (9%) of 23 in CD30+ versus 25 (35%) of 72 in CD30-negative group (P=.02). CD30 expression was not associated with germinal center B-cell-like (GCB) or non-GCB type. Overall survival (OS) was not significantly different between patients with CD30+ and patients with CD30-negative DLBCL, either for all patients or for the subset of patients without MYC rearrangement, regardless of cutoff (P>.05). CD30 expression was not associated with OS in either GCB or non-GCB subtype (P>.05, > 0% cutoff). In conclusion, CD30 expression was detected in up to 25% of cases of DLBCL and was more frequent in tumors without MYC rearrangement. CD30 expression was not associated with OS in R-EPOCH-treated de novo DLBCL patients.

Clinical Significance of TIPE2 Protein Upregulation in Non-Hodgkin's Lymphoma.

Non-Hodgkin's lymphoma (NHL), which includes diffuse large B-cell lymphoma (DLBCL) and peripheral T-cell lymphoma, is a refractory malignant tumor originated from the lymphatic system. TNFAIP8L2 (TIPE2 or tumor necrosis-alpha-induced protein-8 like 2) is a negative regulator for inflammation and an inhibitor for carcinogenesis. However, whether TIPE2 plays a role in lymphomagenesis is unknown. In this study, we determined TIPE2 expression in NHL by immunohistochemistry and investigated its clinicopathological significance in DLBCL. We found that TIPE2 expression was upregulated in both DLBCL and peripheral T-cell lymphoma. But the expression of TIPE2 in T lymphomas was weaker than that in DLBCL. Interestingly, among DLBCL, TIPE2 expression was significantly stronger in the germinal center B-cell (GCB) type than in the non-GCB type. These results suggest that the expression of TIPE2 protein could be a predictor of better prognosis for DLBCL.