Testicular Lymphoma Research Articles

Survey of Long-Term Experiences of Sperm Cryopreservation in Oncological and Non-Oncological Patients: Usage and Reproductive Outcomes of a Large Monocentric Cohort.

Progress in oncological treatment has led to an improved long-term survival of young male cancer patients over the last decades. However, standard cancer treatments frequently implicate fertility-damaging potential. Cryopreservation of sperm is the current standard option to preserve patient’s fertility after treatment, yet long-term data on usage and reproductive experiences is still limited. Natural fertility after treatment and especially in relation to the type of treatment has been poorly analyzed so far. Therefore, we performed a retrospective survey including male patients with an indication for gonadotoxic treatment who cryopreserved reproductive material at our institution between 1994 and 2017. Study questionnaires regarding treatment, material usage, and reproductive outcomes were sent to eligible patients. Additionally, semen analyses of study participants from the time of cryopreservation were evaluated. A total of 99 patients were included in the study. Respondents’ median age was 38.0 years. Most frequent diagnoses were testicular cancer (29.3%) and lymphoma (26.3%). A further 8.1% suffered from autoimmune diseases. Testicular cancer patients had a significantly lower pre-treatment median sperm concentration (18.0 million/ml) compared to non-testicular cancer patients (54.2 million/ml). Until November 2020, the determined sperm usage and cumulative live-birth rate per couple were 17.2% and 58.8%, respectively. Most sperm users received treatments with high (40.0%) or intermediate (33.3%) gonadotoxic potential. 20.7% of all patients reported to had fathered at least one naturally conceived child after treatment, this being the case especially if they had been treated with less or potentially gonadotoxic therapies. In conclusion, our findings emphasize the importance of sperm cryopreservation in the context of male fertility preservation. Furthermore, they indicate that the gonadotoxic potential of patients’ treatments could represent a predictive factor for sperm usage.

MYD88 L265P Augments Proximal B-Cell Receptor Signaling in Large B-Cell Lymphomas Via an Interaction with DOCK8

Diffuse large B-cell lymphoma (DLBCL) is a genetically heterogeneous disease comprised of five subtypes including a subset of poor-prognosis activated B cell (ABC)-enriched tumors with frequent MYD88L265P mutations, often in association with CD79B alterations (Cluster 5 DLBCLs) (Nat. Med. 2018; 24:679-690). Primary central nervous system lymphomas (PCNSLs) and primary testicular lymphomas (PTLs) have similar genetic signatures including recurrent MYD88L265P mutations and concurrent CD79B alterations (Blood 2016; 127: 869-81). These findings prompted us to evaluate a potential role for MYD88L265P in proximal B-cell receptor (BCR) signaling, in addition to its defined function as an intermediary in the Toll-Like Receptor (TLR) pathway and downstream NF-kB activation.In previous studies by Jabara et al., wild-type (WT) MYD88 was found to be constitutively associated with the DOCK8 adapter and the PYK2 tyrosine kinase in normal B-cells (Nat. Immunol. 2012; 13:612-20). In this setting, physiologic ligation of TLR9 with CpG oligodeoxynucleotides (CpG) induced PYK2-mediated phosphorylation of DOCK8, recruitment of Src kinases, including LYN, and downstream activation of the proximal BCR pathway member, spleen tyrosine kinase (SYK) (Nat. Immunol. 2012; 13:612-20). We postulated that mutated MYD88L265P might similarly augment proximal BCR signaling in DLBCLs in the absence of physiologic (CpG-induced) TLR9 signaling. Using three DLBCL cell lines (OCI-Ly1, SU-DHL4 and OCI-Ly7) with intact BCR signaling and WT endogenous MYD88 and CD79B, we first established that physiologic CpG activation of TLR signaling induced the phosphorylation of PYK2 and the proximal BCR signaling components, SYK and Bruton's tyrosine kinase (BTK). Thereafter, we genetically engineered these three DLBCL cell lines to express MYD88 L265P or MYD88 WT, alone or in association with CD79B Y196F. In all three cell lines, the co-expression of MYD88 L265P and CD79B Y196F significantly increased magnitude and duration of SYK and BTK phosphorylation following BCR crosslinking. These findings highlight the likely role of MYD88L265P in CD79BY196F-associated proximal BCR signaling in DLBCL.To elucidate the potential role of the DOCK8 adapter in MYD88 L265P-augmented BCR signaling, we first assessed the colocalization of MYD88 WT or MYD88 L265P with DOCK8 in the same three genetically engineered DLBCL cell lines using proximity ligation assays (PLA), which detect protein-protein interactions at less than 40 nm in situ. In each of these cell lines, we detected significantly increased co-localized MYD88 L265P/DOCK8 signals in comparison to MYD88 WT/DOCK8 signals (p<.0001, all). Additionally, there were significantly increased co-localized DOCK8/LYN signals in DLBCL cell lines that expressed MYD88 L265P rather than MYD88 WT (p<.0001, all). These data provide the first direct evidence of an enhanced association between MYD88 L265P, DOCK8 and LYN in BCR-dependent DLBCLs and a basis for enhanced BCR signaling in primary tumors with concurrent MYD88L265P and CD79B genetic alterations.We next analyzed the consequences of MYD88 L265P-associated, DOCK8-dependent increased proximal BCR signaling by depleting DOCK8 in BCR-dependent DLBCL cells with endogenous MYD88L265P/CD79BY196F alterations (HBL1 and TMD8) or endogenous unmutated MYD88 WT/CD79B WT (OCI-Ly1 and SU-DHL4). ShRNA-mediated DOCK8 knockdown (KD) significantly decreased BCR-mediated phosphorylation of SYK and BTK in MYD88L265P/CD79BY196F DLBCL cell lines but not in lines with MYD88 WT/CD79B WT, highlighting the specific role of DOCK8 in MYD88 L265P-associated proximal BCR signaling. Of great interest, DOCK8 KD selectively decreased the proliferation of MYD88L265P/CD79BY196F, but not MYD88WT/CD79BWT, DLBCLs (p<.004, HBL1 and p<.009, TMD8; p = non sig., OCI-Ly1 and SU-DHL4). Additionally, DOCK8 KD significantly increased the efficacy of chemical PI3Kα/δ (copanlisib) and BTK (ibrutinib) inhibition in MYD88L265P/CD79BY196F DLBCLs (HBL1 and TMD8). Taken together, these data identify DOCK8 as an intermediary in MYD88L265P-driven proximal BCR signaling and a possible treatment target in LBCLs with co-occurring MYD88L265P/CD79BY196F mutations. DisclosuresShipp: AstraZeneca: Consultancy, Research Funding; Immunitas Therapeutics: Consultancy; Bristol Myers Squibb: Research Funding; Merck: Research Funding; Bayer: Other: Institution: Research Grant/Funding; Abbvie: Other: Institution: Research Grant/Funding.

Next-Generation Sequencing Reveals Molecular Heterogeneity in Primary Testicular DLBCL

Background:Primary testicular lymphomas (PTLs) are extra-nodal large B-cell lymphomas (LBCLs) that respond poorly to current empirical treatment options. To date, no PTL whole-exome sequencing analysis based on the Chinese population has been reported. The main objective of this study was to explore the genomic landscape of PTDLBCL, correlating the results with clinical, histological, and immunogenetic features.Methods:We retrospectively analyzed the disease characteristics in a series of 20 patients (13 paired data and 7 unpaired data). In all cases, the diagnosis of PTL was made using appropriate diagnostic criteria for the 2008 WHO classification of lymphoid tumors with combinations of histologic, immunohistochemical, flow cytometric, and genetic evaluation. Medical records were reviewed for demographic and clinical data. We performed whole-exome next-generation sequencing (NGS) and mapped the mutational landscape. Based on the differential gene results, we performed KEGG and GO enrichment analysis, and five potential new pathogenic genes for DLBCL were obtained based on the Polyphen score.Results:A total of 20 DLBCL samples were collected. All samples were subjected to whole-exome sequencing (WES). A standard bioinformatics analysis process was used to identify somatic mutations in each sample, and an average of 1200 somatic mutations was identified, involving an average of 420 mutated genes. (Figure 1.) More than 85% of the somatic mutations were found in only 1 sample, indicating that somatic mutations are highly heterogeneous. In the matched 13 cases, we validated common gene mutations that have been reported, including PIM1 53.84% (7/13), CD79B 38.46% (5/13), CDKN2A 15.38% (2/13), and MYD88 15.38% (2/13). Venn diagram shows that 25% of the mutated genes were specific to the relapsed sample. GO analysis (Figure 2) shows the functional enrichment and differences between relapsed and non-relapsed samples. Non-relapsed samples are enriched in the herpes simplex virus 1 infection pathway. (Figure 3). Focusing on relapsed/non-relapsed samples, we identified 12 mutated genes specific to relapsed samples. Mutation Polyphen scores were calculated to assess mutation deleteriousness. Five potential new pathogenic genes for DLBCL were obtained.Conclusions:Based on whole-sequencing data, we validated the previously reported common genes for PTDLBCL. In addition, we performed GO and KEGG analysis according to the relapsed and non-relapsed groups. In addition, based on the difference in enrichment results, 12 significantly enriched mutated genes in relapsed DLBCL samples are obtained, and mutation Polyphen scores are calculated to assess mutation deleteriousness. And 5 potential new pathogenic genes of DLBCL are identified. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

A case report of testicular diffuse large B-cell malignant lymphoma with cutaneous metastasis: A rare entity.

Introduction and importancePrimary testicular lymphoma (PTL) is a variety of extra-nodal lymphoma taking origin from testis. It accounts 5% of all testicular tumors. Metastasis may occur in contralateral testis, bone, central nervous system and rarely in skin. Herein, we present the case of testicular diffuse large B-cell malignant lymphoma with cutaneous metastasis.Case presentationA 60-year-old male presented with swollen painless solid right testis, with homolateral inguinal nodes. Testicular tumors markers were within normal range. Right radical orchidectomy was performed. Histopathological examination concluded to the diagnosis of Diffuse Large B Cell Lymphoma. Four weeks later, the patient presented alteration of general condition and multiples cutaneous centimetric lesions located in the right inguinal region. Biopsy of this lesion confirmed the diagnosis of metastases from the testicular lymphoma. The patient deceased three days later, before starting further treatment.Clinical discussionPrimary testicular lymphoma is a rare variety of testicular tumors. The prognosis is poor. Metastasis may occur in different sites such as contralateral testis, central nervous system, and skin. The prognosis is usually poor in the rare case of cutaneous metastasis.ConclusionPrimary testicular tumor is an aggressive rare variety of testicular tumors with poor prognosis. Cutaneous metastasis is rarely reported. Cutaneous lesions should be explored and suspected to be malignant. Early treatment with rapid multidisciplinary management is the key for adequate approach.

Testicular FDG Uptake on PET/CT in Patients with Lymphoma: Correlation with Age

The purpose of this observational study was to investigate whether the standard uptake value (SUV) measurement has practical utility in distinguishing secondary testicular involvement from physiologic uptake in patients with lymphoma. A Radiology Information System (RIS) search was conducted for all PET/CT studies performed from 2010-2016 on adult male patients with a diagnosis of lymphoma. Patients with clinical or pathologic diagnosis of testicular lymphoma were excluded to undergo a separate analysis. PET/CT images of 606 patients with 1087 scans, in which 2045 testes were included in the field of view, were reviewed and measurements were performed for standardized uptake values of both testicles (SUVmax) as well as of the liver (SUVmax and SUVmean). The mean SUVmax of the testicles was 3.75 ± 0.90 (range 1.16-8.38). The mean ratio of testis SUVmax / liver SUVmean (T/L) was 1.78 ± 0.43. Trends in SUVmax and age were significant for a negative correlation by a small magnitude of 0.066 per 10 years (P < 0.001). T/L had similar changes with significant low magnitude decrease with increasing age (0.059 per 10-year increase, P < 0.001). In our separate analysis of 3 patients with clinical or pathology proven testicular lymphoma, the average pathologic SUVmax was 13.47 (range 11.39-15.97). This study has the largest known sample size for quantifying physiologic uptake in the testes. SUV measurements to quantify F-18 Fluorodeoxyglucose (FDG) uptake on PET/CT likely have practical utility in discriminating between physiologic and pathologic uptake of FDG in cases of secondary testicular lymphoma.

Hyperosmolar Hyperglycaemic State and Diabetic Ketoacidosis in Nivolumab-Induced Insulin-Dependent Diabetes Mellitus.

Checkpoint inhibitor-associated diabetes can present abruptly with life-threatening complications.Most patients require multiple daily injections of insulin upon discharge.Cessation of checkpoint inhibitor therapy does not revert diabetes.

Testicular Diffuse Large B-Cell Lymphoma-Clinical, Molecular, and Immunological Features.

Simple SummaryTesticular diffuse large B-cell lymphoma (T-DLBCL) is a rare and aggressive lymphoma entity that mainly affects elderly men. It has a high relapse rate with especially the relapses of the central nervous system associating with dismal outcome. T-DLBCL has a unique biology with distinct genetic characteristics and clinical presentation, and the increasing knowledge on the tumor microenvironment of T-DLBCL highlights the significance of the host immunity and immune escape in this rare lymphoma, presenting in an immune-privileged site of the testis. This review provides an update on the latest progress made in T-DLBCL research and summarizes the clinical perspectives in T-DLBCL.Primary testicular lymphoma is a rare lymphoma entity, yet it is the most common testicular malignancy among elderly men. The majority of the cases represent non-germinal center B-cell-like (non-GCB) diffuse large B-cell lymphoma (DLBCL) with aggressive clinical behavior and a relatively high relapse rate. Due to the rareness of the disease, no randomized clinical trials have been conducted and the currently recognized standard of care is based on retrospective analyses and few phase II trials. During recent years, the tumor microenvironment (TME) and tumor-related immunity have been the focus of many tumor biology studies, and the emergence of targeted therapies and checkpoint inhibitors has significantly modulated the field of cancer therapies. Testicular DLBCL (T-DLBCL) is presented in an immune-privileged site of the testis, and the roles of NF-κB pathway signaling, 9p24.1 aberrations, and tumor-infiltrating immune cells, especially immune checkpoint expressing lymphocytes and macrophages, seem to be unique compared to other lymphoma entities. Preliminary data on the use of immune checkpoint inhibitors in the treatment of T-DLBCL are promising and more studies are ongoing.

Limbic Encephalitis with HU-Antibodies in T-cell Anaplastic Lymphoma. A Case Report

Limbic encephalitis is a rare paraneoplastic neurological syndrome usually associated with small cell lung cancers, testicular and breast cancers or B-cell lymphomas. We herein report the first patients with paraneoplastic limbic encephalitis associated with HU antibodies and anaplastic T-cell lymphoma.

The importance of genomic predictors for clinical outcome of hematological malignancies

The importance of genomic predictors for clinical outcome of hematological malignancies

Direct Spread of Primary Testicular Lymphoma along the Gonadal Vessels Detected on F-18 Fluorodeoxyglucose Positron-Emission Tomography/Computed Tomography Imaging.

A 63-year-old male presented with left scrotal swelling and the ultrasound showed a large heterogeneous mass consistent with a testicular malignancy. The patient underwent left-sided orchiectomy which showed diffuse large B-cell lymphoma. The patient was then referred for whole-body F-18 fluorodeoxyglucose positron-emission tomography/computed tomography (FDG PET/CT) imaging which showed multiple hypermetabolic foci extending along the left inguinal canal to the retroperitoneum and the left perinephric space, suggesting direct contiguous spread of the tumor along the gonadal vessels, a form of metastasis unique to primary testicular lymphoma, and demonstrated for the first time on FDG PET/CT imaging.

Low Expression of T/NK-Cell Signature Predicts Poor Survival in Patients with Primary Testicular and Diffuse Large B-Cell Lymphoma

Primary testicular lymphoma (PTL) is a rare and aggressive disease accounting for 1% of malignant lymphomas. Majority of the PTLs are diffuse large B-cell lymphomas (DLBCL). Recently, the impact of tumor microenvironment (TME) and limited immune surveillance on the lymphoma pathogenesis and survival has been recognized. Here, we aimed to characterize cellular and molecular immunological profiles in PTL, and associate the findings with outcome.We used NanoString digital gene expression profiling with the PanCancer Immune panel to assess the expression of 770 immune response genes in tumor tissue of 60 patients with PTL. RNA-sequencing data from a cohort of 96 patients with primary DLBCL was used to validate the findings.Unsupervised hierarchical clustering revealed clusters of genes with differential expression between the PTL patients. The clusters were enriched for genes related to T-cell and natural killer (NK) cell markers and signaling (e.g. CD3, CD4, CD8, ITGB2, PRF1, GZMA/B/H/M/K), as well as cytokine signaling (e.g. IL4, IL9, IL17A/B, TNFSF11, CXCR1, CCL28). Interestingly, the group of patients with low expression of T/NK-cell signature genes had a significantly worse progression-free survival (5-year PFS 33% vs. 65%, p=0.012) and overall survival (5-year OS 33% vs. 67%, p=0.004), as compared to the other patients. In Cox multivariate analysis with international prognostic index (IPI), T/NK-cell signature retained independence for prediction of better PFS and OS (RR=0.356, p=0.014 and RR=0.306, p=0.006, respectively). The signature was identified also in a cohort of 96 patients with primary DLBCL, but not in Hodgkin lymphoma. Multiplex immunohistochemistry confirmed lower percentage of CD3+, CD3+CD4+, and CD3+CD8+ lymphocytes in the tumor tissue of the group of PTL patients with poor prognosis (11.8% vs. 24.1%, 5.0% vs. 15.6%, and 4.6% vs. 12.3%, respectively), and association of their higher amount with favorable PFS and OS. Moreover, the RNA level expression of MHC class I and II genes was significantly lower in the poor prognosis group.In conclusion, our results demonstrate that loss of T/NK cell signature and MHC class I and II gene expression are associated with high risk of recurrence and death in patients with PTL and DLBCL. They further highlight the importance of the immune escape as a mechanism of treatment resistance. DisclosuresPollari:Takeda: Other: Conference attendance fees and travel expences; BMS: Other: Conference attendance fees and travel expences; Novartis: Other: Conference attendance fees and travel expences; Pierre Fabre: Other: Conference attendance fees and travel expences; Roche: Consultancy. Mannisto:Takeda: Honoraria, Other: Travel expence; Roche: Honoraria, Other: Travel expence; Amgen: Other: Travel expence; Novartis: Other: Travel expence; Celgene: Other: Travel expence; Gilead: Other: Travel expence; Pfizer: Honoraria; SOBI: Honoraria. Mustjoki:BMS: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; Pfizer: Honoraria, Research Funding; Ariad: Research Funding; Celgene: Honoraria. Leppa:Roche: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding; Bayer: Research Funding; Merck: Consultancy, Honoraria; Janssen Cilag: Consultancy, Research Funding.

Stage I Lymphoma: Difference in Survival Outcome By Site of Involvement

Background: Less than half of patients with lymphoma present with stage I or II disease and this presentation may be nodal or extranodal. Stage I lymphoma is commonly treated with chemotherapy with or without radiation therapy (RT), however little is known if primary site of involvement and treatment modalities affect survival outcome.Patients and Methods: TheSurveillance, Epidemiology and End Results (SEER) database was used to evaluate survival outcomes among adult (≥18 yrs) patients diagnosed with stage I lymphoma between 1998 and 2014. We excluded histologic subtypes that develop in extranodal sites such as cutaneous T-cell lymphoma, enteropathy-associated T-cell lymphoma, hepatospenic T-cell lymphoma and extranodal NK-cell lymphoma. We also excluded central nervous system and testicular lymphoma. Overall survival (OS) was calculated from diagnosis to death from any cause using the Kaplan-Meier method. Differences in the survival function between groups were analyzed using the log-rank test. Cox proportional hazards models were used to evaluate associations between patient characteristics and survival.Results: A total of 72095 patients were diagnosed with stage I disease during the study period. The median age of the patients was 66 years (range, 18-104 years). After excluding diseases as described above, diffuse large B-cell lymphoma (DLBCL) was the most common histology (N=24,574), followed by follicular lymphoma (FL, N=13,061), marginal zone lymphoma (MZL, N=9,893), Hodgkin lymphoma (HL, N=5,325) and peripheral T-cell lymphoma (PTCL, with PTCL not otherwise specified, anaplastic large cell lymphoma and angioimmunoblastic T-cell lymphoma, N=2,071). HL was excluded from the analysis with few extranodal diseases. With a median follow up of 69 months, the median OS was 104, 179, 165 and 106 months in patients with DLBCL, FL, MZL and PTCL, respectively. Thirty-seven percent of patients received resection of primary tumor. Thirty-nine percent of patients received neither RT (40% of them received surgical resection) nor chemotherapy, 15% of patients received RT alone, 29% of patients received chemotherapy alone and 17% of patients received combined modality therapy (CMT).Hazard ratios (HR) for OS in stage I extranodal disease compared to stage I nodal disease in each histologic subtype are summarized in Table. Skin origin was associated with longer OS in all histologic subtypes. Breast origin was associated with longer survival in DLBCL and FL. Thyroid origin was associated with longer survival in FL and MZL. Stomach origin was associated with shorter survival in DLBCL, FL and PTCL but not in MZL. Small intestine origin was associated with longer survival in FL but associated with shorter survival in PTCL. Bone origin was associated with shorter survival in FL and PTCL. The difference in HR for site of involvement among histologic subtype is significant; overall, compared to nodal stage I disease, extranodal stage I disease was associated with longer survival in MZL but associated with shorter survival in PTCL.CMT was significantly associated with longer survival in DLBCL while RT alone showed similar survival outcome in FL, MZL and PTCL (Figure). Local control by surgical resection was also associated with significantly longer survival in all histologic subtypes; HR for surgical resection was 0.81 (p<0.001) in DLBCL, 0.83 (p<0.001) in FL, 0.87 (p=0.002) in MZL and 0.85 (p=0.033) in PTCL.Summary: There was a significant difference in survival outcome in stage I lymphoma by site of involvement. RT showed significant impact in survival outcome for patients with stage I FL, MZL and PTCL. [Display omitted] DisclosuresFanale:AMGEN: Membership on an entity's Board of Directors or advisory committees; GENENTECH: Research Funding; MOLECULAR TEMPLATES: Research Funding; TAKEDA: Honoraria, Research Funding; SEATTLE GENETICS: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; BMS: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; MERCK: Membership on an entity's Board of Directors or advisory committees, Research Funding; ONYX: Research Funding; ADC THERAPEUTICS: Research Funding; CELGENE: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding. Westin:Apotex: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees; Kite Pharma: Membership on an entity's Board of Directors or advisory committees; Novartis Pharmaceuticals Corporation: Membership on an entity's Board of Directors or advisory committees. Nastoupil:Abbvie: Honoraria, Research Funding; Janssen: Honoraria, Research Funding; Celgene: Honoraria, Research Funding; Genentech: Honoraria, Research Funding; Karus Therapeutics: Research Funding; TG Therapeutics: Honoraria, Research Funding; Gilead: Honoraria. Neelapu:Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Merck: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Poseida Therapeutics, Inc: Research Funding; Cellectis Inc.: Research Funding; Karus: Research Funding; Bristol-Myers Squibb: Research Funding; Kite Pharma: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding.

Clinical Characteristics and Outcome of Diffuse Large B-Cell Lymphoma with 9p24.1 Copy Number Alterations

Introduction: Copy number alterations (amplifications or gains, CNAs) and/or translocations involving 9p24.1 have been reported in classical Hodgkin lymphoma (cHL), primary mediastinal large B-cell lymphoma (PMBL), Epstein-Barr virus (EBV)-negative primary central nervous system lymphoma (PCNSL), and primary testicular lymphoma (PTL). These alterations often lead to over-expression of PD-L1, PD-L2, and JAK2, providing the rationale for immunotherapy targeting PD-1/PD-L1. Immune checkpoint inhibitors nivolumab and pembrolizumab have proven to be efficacious in treating relapsed and/or refectory cHL. While there is evidence that 9p24.1 CNA is also present in de novo diffuse large B-cell lymphoma (DLBCL), there is little known about its prevalence, association with other molecular events, or its impact on clinical outcome.Methods: Using the Molecular Epidemiology Resource of the University of Iowa/Mayo Clinic lymphoma SPORE, 200 patients with newly diagnosed DLBCL from May 2002 to September 2012 and treated with standard immunochemotherapy were included in this study. All patients were prospectively followed for outcomes. Copy number analysis was performed using OncoScan Whole Genome FFPE Array for 169 cases and whole exome sequencing (WES) for 31 cases. Raw sample files from both analyses were loaded into the Nexus Copy Number Software (V8, BioDiscovery, El Segundo, CA) and analyzed for 9p24.1 CNA. Event-free survival (EFS) was defined as time from diagnosis to progression or relapse, unplanned re-treatment after initial immunochemotherapy, or death from any cause. Overall survival (OS) was defined as time from diagnosis to death from any cause. Categorical data were analyzed by Chi-square test, and time-to-event data were analyzed using Kaplan-Meier and Cox proportional hazards models. Statistical analysis was done in SPSS (V22, IBM, Armonk, NY).Results: Overall, 9p24.1 CNA was detected in 16 (8%) of the 200 patients. Twenty-four cases had available RNASeq data; those with 9p24.1 CNA had significantly increased expression of PD-L1 when compared to cases with normal 9p24.1 (6.4 vs 2.6 log2 RPKM, P < 0.001). Clinical characteristics of patients with and without 9p24.1 CNA are summarized in Table 1. Compared to those without 9p24.1 CNA, patients with 9p24.1 CNA were younger, with a median age of 54.5 (range 28-78) vs 64 (range 26-89) years (P= 0.024, Mann-Whitney U test). Patients with 9p24.1 CNA appeared to have more extranodal involvement, trended towards the ABC subtype, and had less bone marrow involvement, although not statistically significant (Table 1). After a median follow up of 93.1 (range 19.0-169.6) months, there were a total of 95 events and 74 deaths in the entire cohort. Median EFS was not reached for patients with 9p24.1 CNA, and was 8.6 years (95% CI 5.4-11.8) for those without (Figure 1). The 2-year EFS rates were 68.8% vs 67.8% for patients with and without 9p24.1 CNA, respectively, and the 5-year EFS rates were 68.8% vs 57.9%. The 5-year OS rates were 68.8% vs 70.5% for patients with and without 9p24.1 CNA, respectively. After adjusting for IPI score and cell of origin, 9p24.1 CNA was associated with superior EFS (HR = 0.232, 95% CI = 0.054-0.996, P= 0.049) and a trend of superior OS (HR = 0.320, 95% CI = 0.073-1.392, P= 0.129).Conclusions: 9p24.1 CNA was present in 8% of de novo DLBCL patients and was associated with increased expression of PD-L1. Patients with 9p24.1 CNA were younger compared to those without. Presence of 9p24.1 CNA was associated with statistically significant superior EFS. While immune checkpoint inhibitors targeting PD-1/PD-L1 are being evaluated in DLBCL in clinical trials, whether 9p24.1 CNA is a biomarker predicting favorable response and treatment outcome should be further examined. [Display omitted] DisclosuresWitzig:Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; Kura: Research Funding; Acerta: Research Funding; Novartis: Research Funding. Ansell:Affimed: Research Funding; Seattle Genetics: Research Funding; Celldex: Research Funding; Merck: Research Funding; Bristol-Myers Squibb: Research Funding. Nowakowski:Genetech: Consultancy, Research Funding; Nanostring: Research Funding; Bayer: Consultancy, Research Funding; pharmacyclics: Consultancy; Abbvie: Consultancy; celgene: Consultancy, Research Funding; Morphosys: Consultancy, Research Funding. Cerhan:Janssen: Other: Scientific Advisory Board (REMICADELYM4001); Janssen: Other: Multiple Myeloma Registry Steering .

High Amount of PD-L1+ Tumor Associated Macrophages Is Associated with Better Survival in Patients with Primary Testicular Lymphoma

Introduction:Primary testicular lymphoma (PTL) is a rare and aggressive extranodal lymphoid malignancy with poor outcome to therapy and largely uncharacterized biology. Majority of PTLs are diffuse large B-cell lymphomas (DLBCL). Tumor associated macrophages (TAMs) are known to have prognostic impact in DLBCLs, but their role in PTLs has not been addressed. The aim of this study was to examine the association of TAMs with disease characteristics and survival in patients with PTL.Patients and methods:We identified 74 patients with PTL, all representing DLBCL histologically. Tissue microarray (TMA) was constructed from PTL samples and TMA slides were stained using multiplex immunohistochemistry (mIHC), allowing a detection of four markers (CD68, cMAF, PD-L1 and PD-L2) and nuclei simultaneously. Expression profiles were analyzed quantitatively with the image analysis software Cell Profiler. In addition, CD68, PD-L1, and PD-L2 mRNA levels were measured from 60 PTL samples.Results:For the 74 patients in the mIHC cohort, median age at diagnosis was 70 years (36-92 years) and median follow-up 67 months (range 6.7 to 120 months). Majority of the patients represented non-germinal center B-cell (non-GCB) phenotype (76%), low stage (I-II, 64%), and low/intermediate International Prognostic Index (IPI 0-2, 68%). Thirty-five patients (47%) were treated with rituximab containing chemotherapy and 36 patients (49%) received central nervous system (CNS) prophylaxis, most of them (n=34) with CNS penetrating intravenously administered chemotherapy. Altogether 34 deaths, 24 relapses and 24 lymphoma associated deaths occurred during the follow-up. 5-year overall survival (OS), disease specific survival (DSS) and progression free survival (PFS) rates were 56%, 68%, and 53%, respectively.The median proportions of PD-L1+ cells, CD68+, PDL1+CD68+, CD68+cMAF+ (M2 type), and PD-L1+CD68+cMAF+ macrophages of all cells in the tumor tissue were 15.3%, 23.7%, 9.1%, 1.3%, 0.5%, respectively. In univariate analyses with continuous variables, the proportion of PD-L1+ cells (p=0.039) and PDL1+CD68+ macrophages (p=0.043) correlated with a favorable OS. A trend towards a better OS was also observed for PD-L1+CD68+cMAF+ (p=0.090) macrophages, whereas neither PD-L1-CD68+cMAF+ macrophages nor PD-L1+CD68- cells had an impact on survival. Furthermore, no correlation with survival was found for any of the PD-L2+ cells. According to Kaplan-Meier estimates, the patients with the high proportion of PD-L1+CD68+ macrophages had a 5-year OS of 65% in comparison to 41% for those with the lower counts (p=0.026). The difference was more clear in the subgroup of patients with a stage III-IV disease (n=24, 5-year OS, 67% vs 25%, p=ns). When clinical characteristics of the patients were compared according to PD-L1+CD68+ cell counts, no significant differences in age or molecular subtype were observed between the subgroups. However, in the high PDL1+CD68+ group, more patients had stage I-II disease (p<0.001) and low/intermediate IPI score (IPI 0-2, p=0.023). PD-L1 and CD68 gene expressions correlated with corresponding cell counts (for both, rs≥0.673, p<0.001) and favorable OS (PD-L1, p=0.007; CD68, p=0.016).Conclusion:Our data demonstrate a significant variation in TAM contents and their phenotypes in PTL, and an association of PD-L1+ TAMs with more localized disease and favorable survival in patients with PTL. DisclosuresPollari:Novartis: Other: Conference attendance fees and travel expences; Pierre Fabre: Other: Conference attendance fees and travel expences; BMS: Other: Conference attendance fees and travel expences; Takeda: Other: Conference attendance fees and travel expences; Roche: Consultancy. Mannisto:Takeda: Honoraria, Other: Travel expence; Amgen: Other: Travel expence; Novartis: Other: Travel expence; Celgene: Other: Travel expence; Gilead: Other: Travel expence; Pfizer: Honoraria; Roche: Honoraria, Other: Travel expence; SOBI: Honoraria. Mustjoki:BMS: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; Pfizer: Honoraria, Research Funding; Celgene: Honoraria; Ariad: Research Funding. Leppa:Takeda: Consultancy, Honoraria, Research Funding; Janssen Cilag: Consultancy, Research Funding; Roche: Consultancy, Honoraria, Research Funding; Merck: Consultancy, Honoraria; Bayer: Research Funding.

Th1/2 Immune Response Signature Predicts Outcome after Dose-Dense Immunochemotherapy in Patients with High Risk Diffuse Large B-Cell Lymphoma - Results from Nordic Lymphoma Group Trials

Introduction: Despite better therapeutic options and improved survival of diffuse large B-cell lymphoma (DLBCL), 30-40% of the patients still relapse and have dismal prognosis. Recently, the impact of genomic aberrations, allowing lymphoma cells to escape immune recognition on DLBCL pathogenesis has been recognized. However, whether immune related signatures could be used as determinants for treatment outcome has not been rigorously evaluated. Here, our aim was to elucidate the immunologic characteristics of the tumor microenvironment, and associate the findings with outcome in patients with high-risk DLBCL.Patients and methods: We specifically examined gene expression in the tumor microenvironment, using RNA isolated from formalin fixed paraffin-embedded (FFPE) tumor samples from 81 patients with DLBCL. Gene expression analysis was conducted on the NanoString nCounter gene expression platform using a 770-gene PanCancer Immune panel. The predictive impact of the gene expression signatures was assessed using a cohort of 65 high risk DLBCL patients enrolled in the Nordic CHIC trial (Leppä et al., 2016). The findings were validated using exon array data from an independent cohort of 38 high risk DLBCL patients treated in the Nordic CRY-04 trial (Holte et al., 2013; Leivonen et al., 2017a). In both trials, the patients received dose-dense immunochemotherapy (R-CHOEP-14) and systemic central nervous system prophylaxis (HD-Mtx and HD-Ara-C).Results: Unsupervised hierarchical clustering revealed three major clusters of genes with differential expression between the patients. The clusters were enriched for the genes related to focal adhesion and extracellular matrix (ECM) receptor interactions (eg. COL3A1, ITGA, ITGB, VEGFC, VEGFA, PDGFRB, PDGFC, FN1 ), T/NK-cell signaling (eg. ICAM1, CD3, CD4, CD8 GZMB, ITGB2 ), and type 1/2 immune responses (eg. IL4, IFNL1, IFNL2, IL13, IL24 ). Interestingly, type 1/2 immune response signature correlated with survival. In the CHIC cohort, the group of patients (n=10) with high expression of type 1/2 immune response signature genes had a 4-year PFS of 60% in comparison to 91% for those (n=55) with low expression (p = 0.021). The corresponding 4 y OS rates were 64% vs 90% (p=ns). The risk of progression was 4.0 fold higher for the patients with high expression of type 1/2 immune response signature genes (95% CI 1.120-14.280, p=0.033). No differences were observed in sex, age, stage, IPI scores, or molecular subgroups between the low and high type 1/2 immune response subgroups. The prognostic impact of the signature was confirmed in the CRY-04 exon array cohort (4-y PFS; 46% vs 80%, p = 0.021, 4-y OS 62% vs 88%; p = 0.014). In the CRY-04 cohort, the risk of progression was 3.6 fold (95% CI 1.13-11.20, p=0.030) and death 4.8 fold (95% CI 1.20-19.26, p=0.027) higher for the group of patients with high expression of the type 1/2 immune response signature genes. In contrast to our data on primary testicular lymphomas (PTLs) (Leivonen et al., 2017b), T/NK cell signature did not correlate with outcome in either of the trial cohorts. Correspondingly, type 1/2 immune signature was not associated with outcome in the cohorts of PTL or DLBCL patients treated with conventional R-CHOP regimen.Conclusion: Our data suggest that immune related signatures exhibit treatment-specific roles in diffuse large B-cell lymphoma. For the high risk DLBCL patients treated with dose-dense immunochemotherapy, high expression of type 1/2 immune response signature genes predicts a poor outcome. A detailed characterization of immune cell composition in the tumor microenvironment and its impact on survival is ongoing. DisclosuresJørgensen:Roche: Research Funding. Holte:Novartis: Consultancy, Honoraria; Celgene: Consultancy; Janssen Cilag: Consultancy; Nordic Nanovector: Consultancy; Mundipharma: Consultancy. Leppa:Merck: Consultancy, Honoraria; Roche: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding; Janssen Cilag: Consultancy, Research Funding; Bayer: Research Funding.

Ineffectiveness of high-dose methotrexate for prevention of CNS relapse in diffuse large B-cell lymphoma.

Central nervous system (CNS) relapse affects 5% of diffuse large B-cell lymphoma (DLBCL) patients and portends a poor prognosis. Prophylactic intravenous high-dose methotrexate (HD-MTX) is frequently employed to reduce this risk, but there is limited evidence supporting this practice. We conducted a multicenter retrospective study to determine the CNS relapse risk with HD-MTX in DLBCL patients aged 18-70 years treated in Alberta, Canada between 2012 and 2019. Provincial guidelines recommended HD-MTX for patients at high-risk of CNS relapse based upon CNS-IPI score, double-hit lymphoma, or testicular involvement. Among 906 patients with median follow-up 35.3 months (range 0.29-105.7), CNS relapse occurred in 1.9% with CNS-IPI 0-1, 4.9% with CNS-IPI 2-3, and 12.2% with CNS-IPI 4-6 (p < .001). HD-MTX was administered to 115/326 (35.3%) high-risk patients, of whom 96 (83.5%) had CNS-IPI score 4-6, 45 (39.1%) had double-hit lymphoma, and four (3.5%) had testicular lymphoma. The median number of HD-MTX doses was two (range 1-3). Central nervous system relapse risk was similar with versus without HD-MTX (11.2% vs. 12.2%, p = .82) and comparable to previous reports of high-risk patients who did not receive CNS prophylaxis (10-12%). In multivariate and propensity score analyses, HD-MTX demonstrated no association with CNS relapse, progression-free survival, or overall survival. This study did not demonstrate a benefit of prophylactic HD-MTX in this high-risk patient population. Further study is required to determine the optimal strategy to prevent CNS relapse in DLBCL.

Different chemokine profile between systemic and testicular diffuse large B-cell lymphoma

Although treatment for diffuse large B-cell lymphoma (DLBCL) has taken some notable steps in the 2000s, there are still subgroups of patients suffering from high mortality and relapse rates. To further improve treatment outcomes, it is essential to discover new mechanisms of chemotherapy resistance and create new treatment approaches to overcome them. In the present study, we analyzed the expression of chemokines and their ligands in systemic and testicular DLBCL. From our biopsy sample set of 21 testicular and 28 systemic lymphomas, we were able to demonstrate chemokine profile differences and identify associations with clinical risk factors. High cytoplasmic CXCL13 expression had correlations with better treatment response, lower disease-related mortality, and limited stage. This study suggests that active CXCR5/CXCL13 signaling could overtake the CXCR4/CXCL12 axis, resulting in a better prognosis.

Fine Needle Aspiration Cytology of Primary and Secondary Testicular Lymphoma and its Clinical Mimickers- A Definite Indication

Albeit being rare, the testicular lymphoma is the most common testicular malignancy in elders. In early stages, its symptoms are non-specific and can mimic other underlying diseases. The available knowledge of cytology on this entity is not adequate. In case when a clinician comes across a testicular enlargement case, a decision has to be made when to send the patient for FNAC. Similarly, the pathologist who is assessing the patient should also make a decision whether to perform Fine Needle Aspiration Cytology (FNAC) on the patient or not, and what morphology is expected on cytological smears to be able to confidently call it testicular lymphoma. We have undertaken this study to serve the purpose of enhancing the sparse knowledge available on this disease, dealt with its multiorgan involvement, and explained how FNAC is a valuable utility in testicular enlargement cases in which serological markers of testicular malignancy are negative.

Primary Testicular Non-Hodgkin Lymphoma: A Retrospective Single Centre Experience of 26 Cases with Long Follow Up

Introduction: Primary testicular lymphoma is a rare and aggressive malignancy representing 1% to 2% of all non-Hodgkin lymphomas (NHL) and accounts for approximately 5% of all testicular tumors. The aim of this study was to analyse clinical characteristics, therapy and survival outcomes of patients with primary testicular lymphoma diagnosed at our hospital from 1998 to 2017, and to compare differences in survival based on Ann Arbor Stages. Methods: A retrospective patient chart review was done to analyse the patient clinical characteristics, therapy and survival outcomes. Survival was calculated using Kaplan-Meier survival analysis. Results: Twenty-six patients were included; the mean age was 85±6.7 years. All patients presented with testicular tumor. Of the 26 patients, 17 patients were stage I, one stage III, and 8 stage IV. Orchiectomy was performed in all patients. 8 patients received no further treatment. Of the remaining 18 patients, overall, 18 received systemic chemotherapy and 5 radiation therapy. Six patients received intrathecal chemotherapy prophylaxis. Seventeen patients achieved complete remission. Four patients achieved partial remission; one patient had no response. Eleven patients died. Median survival was 144 months and was statistically significantly different between early versus advanced stage (stage I/II: 144 months; stage III-IV: 73 months, p=0.006). Conclusion: Primary testicular lymphomas are a rare and aggressive malignancy of extranodal lymphomas. Survival was different in early stages I/II versus advanced stages III- IV. Randomized, multi-center prospective studies could help to establish better prognostic and multi-modal therapy strategies.

Testicular Lymphoma manifesting as neurological symptoms due to secondary neurolymphomatosis: A case report

Neurolymphomatosis is a rare manifestation of lymphoma presenting as diffuse invasion and involvement of peripheral and spinal nerves. Due to the common presenting symptomatology of neurologic complaints localizing to the affected peripheral nerve, lymphoma as the underlying etiology can be diffificult to diagnose. Here we present the case of a gentleman presenting with right extremity neuropathic symptoms, subsequently discovered to have diffuse large B-cell lymphoma of testiticular orgin after nerve biopsy revealed neurolymphomatous involvement of a spinal nerve. This case highlights the importance of the consideration of neurolymphomatosis in the work up of neuropathic symptoms, as well as the full assessment for the site of primary involvement.