Hodgkin Lymphoma Tumor Research Articles

Synchronous Hodgkin's Lymphoma and Seminoma, a Rare Co-Existence and an Important Lesson

Introduction:Second malignancies, including lymphoma, occur at a higher incidence in men previously treated for germ cell tumors than in the general population (1). Synchronous presentation of seminoma and lymphoma is rare but has important ramifications for the treatment of both malignancies. Without clinical vigilance this situation may be easily missed, leading to inappropriate management of each cancer. We describe a patient found to have synchronous seminoma and Hodgkin Lymphoma and discuss the effects of the dual diagnoses on his evaluation and care.Case presentation:A 59 year old male with no significant medical history presented with progressive swelling and erythema of the right testis. Testicular cancer was suspected and he underwent a radical right inguinal orchiectomy. Pathology revealed a 5.7cm seminoma of the testis with lymphovascular invasion and without spermatic cord involvement (pT2) (Fig. 1). His tumor markers including AFP, LDH, and Beta-HCG were normal (S0). A CT scan of the chest, abdomen and pelvis followed by a PET/CT revealed enlarged, hypermetabolic mediastinal, hilar and periportal lymphadenopathy interpreted by the radiologist as concerning for metastatic disease. Given the atypical distribution for lymphadenopathy from testicular seminoma, an excisional biopsy of a left hilar node was performed and revealed Classical Hodgkin Lymphoma with IHC positive for CD15, CD30 and PAX-5 (Fig 2). He denied any B-symptoms and his bone marrow was uninvolved by lymphoma (stage IIIA). Adjuvant therapy for his germ cell tumor, otherwise an important consideration, was deferred and he began chemotherapy with adriamycin, bleomycin, vinblastine and dacarbazine (ABVD) for 6 cycles. Interim PET/CT after 2 cycles of ABVD showed a complete response. He has completed 6 cycles of ABVD and chose observation as opposed to single dose of adjuvant carboplatin for his seminoma and is currently under surveillance for both malignancies.Discussion:The unusual coexistence of Hodgkin Lymphoma and seminoma has rarely been documented in medical literature, with three other cases previously reported (2, 3). In each case a biopsy of lymphadenopathy, primarily outside the retroperitoneum, yielded a diagnosis of lymphoma. Both Hodgkin Lymphoma and germ cell tumors commonly involve lymph nodes and present in young men. Lymphadenopathy may understandably be assumed to represent metastatic disease in a young man with known testicular cancer. Clinical vigilance is necessary to question the nature of atypical sites of lymphadenopathy in such a patient, and to pursue the possibility of an alternate diagnosis with a lymph node biopsy. A missed diagnosis of lymphoma in such a patient would also mean harmful over-staging of the germ cell tumor. While these two cancers represent two of the most curable malignancies, their treatment is different and would be grossly wrong if each cancer is not correctly diagnosed and staged.Conclusion:Our report highlights the importance of clinical suspicion of a lymphoma in patients with another cancer and lymphadenopathy not typical of metastatic disease for that tumor type. In such situations a lymph node biopsy is crucial in order to proceed with the correct therapy of each malignancy. While the simultaneous presentation of Hodgkin Lymphoma and seminoma is rare, cases like ours highlight the importance of questioning metastatic disease when lymphoma seems to be a possibility.

High-risk interim PET negative patients in Hodgkin's lymphoma

Interim FDG-PET/CT done as early as after the first or second cycle of first line chemotherapy is now recognised as a good predictor of outcome in classic Hodgkin's lymphoma. This scan has been successfully used as a guide to escalate or de-escalate therapy in several important trials. 1 Radford J Illidge T Barrington S PET-directed therapy for Hodgkin's lymphoma. N Engl J Med. 2015; 373: 392 Crossref PubMed Scopus (4) Google Scholar Interim PET negative scans have a high negative predictive value, the abrupt decrease of the fluorodeoxyglucose signal expression during therapy is explained by the switch-off of activity of the microenvironmental cells recruited by chemokines that are produced by the Hodgkin Reed-Sternberg cells. 2 Gallamini A Positron emission tomography scanning: a new paradigm for the management of Hodgkin's lymphoma. Haematologica. 2010; 95: 1046-1068 Crossref PubMed Scopus (31) Google Scholar Indeed, the intense fluorodeoxyglucose uptake observed in Hodgkin's lymphoma at staging of the disease is explained mainly by the activity of the GLUT3 receptors on the microenvironmental cells, which account for 99% of cells in the Hodgkin's lymphoma tumour. However, 10–15% of patients with a negative interim PET scan still had disease progression or relapsed. 3 Johnson P Federico M Kirkwood A et al. Adapted treatment guided by interim PET-CT Scan in advanced Hodgkin's lymphoma. N Engl J Med. 2016; 374: 2419-2429 Crossref PubMed Scopus (488) Google Scholar In a prospective study, 3 Johnson P Federico M Kirkwood A et al. Adapted treatment guided by interim PET-CT Scan in advanced Hodgkin's lymphoma. N Engl J Med. 2016; 374: 2419-2429 Crossref PubMed Scopus (488) Google Scholar the 3 year progression-free survival rate of patients who were PET negative after two cycles of ABVD (doxorubicin, bleomycin, vinblastine, and dacarbazine) was 86% (95% CI 82·1–88·6). No diagnostic tool has been proposed yet for better identification of PET negative patients who relapse these patients. The combined role of biomarkers and interim PET scan in prediction of treatment outcome in classical Hodgkin's lymphoma: a retrospective, European, multicentre cohort studyThe CART algorithm correctly predicted the response to treatment in more than a half of patients who had a relapse or disease progression despite a negative PET-2 scan, thus increasing the negative predictive value of PET-2. In keeping with preliminary results from interim PET response adapted clinical trials of patients with advanced Hodgkin's lymphoma, there might be a non-negligible proportion of treatment failures in the interim PET negative group treated with standard ABVD. Full-Text PDF

CD58 mutations are common in Hodgkin lymphoma cell lines and loss of CD58 expression in tumor cells occurs in Hodgkin lymphoma patients who relapse.

CD58 is involved in immune recognition of tumor cells via binding of the CD2 receptor expressed on cytotoxic T cells. In diffuse large B-cell lymphoma, mutations of the CD58 gene are reported to contribute to immune evasion of the tumor cells. We previously showed CD58 mutations in three Hodgkin lymphoma (HL) cell lines by whole-exome sequencing. In this study, we confirmed the mutations by Sanger sequencing at the DNA and RNA level and showed low levels or total loss of CD58 mRNA expression in two of the three cell lines. CD58 protein expression as determined by flow cytometry, western blotting and immunohistochemistry was absent in all three mutated HL cell lines. In primary tissue samples, loss of CD58 expression was observed in 11% of the patients who relapse. These data suggest that loss of CD58 is a potential immune escape mechanism of HL tumor cells, especially in clinically aggressive disease.

Abstract 3405: The analysis of TRPV6 expression in lymphoid tumours

Abstract Introduction: TRPV6 is a calcium channel that is found overexpressed in various malignancies and is correlated with the prognosis of patients with prostate cancer. TRPV6 targeted anticancer therapies are in development. We are interested in the potential of TRPV6 as a potential biomarker and therapeutic target in lymphoma. This retrospective study examines the expression levels of TRPV6 in various lymphoid tumour types, and correlates expression levels with grade, prognostic scores, and survival rates. Methods: A clinical-pathological database was constructed using the health records and lymphoid tumor samples of patients diagnosed with diffuse large B-cell lymphoma (DLBCL), Follicular lymphoma (FL), small lymphocytic lymphoma/chronic lymphocytic leukemia (SLL/CLL), and Hodgkin's lymphoma (HL). Immunohistochemical studies were performed on lymphoid tumor samples to analyze the level of TRPV6 expression. Tumor samples were graded on a 3-point scale. A score of +3 indicated dense staining with high expression levels, and a score of +1 indicated minimal staining with low expression levels. TRPV6 expression levels were correlated with prognostic scores (IPI, FLIPI) and survival rates using Chi-squared, Mantel-Cox, and Kaplan-Meier survival curves. Descriptive statistics were used to describe patient demographics. Results: We found high TRPV6 expression levels in DLBCL and HL tumor samples with > 40% of samples scoring +3. We found low TRPV6 expression levels in FL and SLL/CLL with > 40% of samples scoring +1. We found no significant correlation between the level of TRPV6 expression and prognostic scores or survival rates in any of the lymphoma subtypes studied. In follicular lymphoma tumor samples, it was noted on observation that the large tumor cells generally stained +3 while the small tumor cells generally stained +1. Conclusion: This retrospective study showed DLBCL and HL tumors have high levels of TRPV6 expression. It also showed that TRPV6 is found highly expressed in the large transformed cells (centroblasts) in follicular lymphoma. These results demonstrate a need to further assess the role of TRPV6 in the aggressiveness of follicular lymphoma, as well as the prognosis and survival in DLBCL and HL with a larger sample size. Further studies of the potential of TRPV6 as a therapeutic target in lymphoma will focus on DLBCL, high grade/transformed FL, and HL. Citation Format: Ashley M. DiPasquale, Tarek Rahmeh, Lauren Andrew, Jane Agar, Kim Miller, Matthew Finniss, Alli Murugesan, Anthony Reiman. The analysis of TRPV6 expression in lymphoid tumours. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3405.

Abstract 2323: Immune checkpoint inhibition by anti-PD-1 or CD137 co-stimulation enhances cytotoxicity towards CD30+ tumors mediated by the bispecific tetravalent CD30/CD16A TandAb AFM13

Abstract AFM13 is an NK-cell engaging CD30/CD16A bi-specific tetravalent TandAb antibody currently in Phase 2 clinical development in Hodgkin Lymphoma (HL) and CD30+ malignancies. Immune checkpoints inhibitors have demonstrated clinical efficacy in a variety of cancers, including HL. NK-cells are also regulated by a number of check-points, prompting us to investigate the combination of AFM13 with several immuno-modulatory antibodies. In previous experiments we were able to demonstrate higher efficacy of AFM13 than several immuno-modulatory antibodies in monotherapy and strong synergy between AFM13 and an anti-PD-1 antibody in vitro, as well as in vivo PDX models with human CD30+ HL tumors. In order to investigate the underlying immunological mechanisms we employed the same PDX model by implanting tumor fragments derived from surgical specimens of HL patients in immuno-deficient mice. After establishing tumors, mice were reconstituted with autologous patient-derived PBMC and treated with AFM13 alone and in combination with anti-PD-1 weekly for a total of three weeks. Tumor size, tumor-infiltrating human lymphocytes, myeloid cells and intra-tumoral cytokines were evaluated on days 30, 44, and 58, i.e. 2, 16 and 30 days after treatment start. While monotherapy with AFM13 was reproducibly more potent than anti-PD-1, significant synergy was observed when both agents were combined. Analysis of the tumors on day 58 revealed a strong correlation between tumor growth inhibition and levels of tumor-infiltrating NK-cells, T-cells, myeloid cells and intra-tumoral cytokines such as IFNg. In contrast to anti-PD-1 monotherapy, which only induced T-cell infiltration, AFM13 monotherapy was able to induce infiltration of NK- and T-cells in the tumors, however the combination further enhanced infiltration of both, NK- and T-cells. AFM13 resulted in stronger infiltration of macrophages than anti-PD-1, which was also increased by the combination of both agents, therefore further supporting cross-talk between innate and adaptive immunity. Furthermore, tumor analyses at earlier time-points (days 30 and 44) showed that the initial immune response is characterized by NK-cell infiltration and activation, as well as infiltration of macrophages, whilst the adaptive immune response by T-cells and activated dendritic cells was more pronounced on day 58. Combining AFM13 and anti-PD-1 augments infiltration and activation of all immune subpopulations. In conclusion, our data support strong synergistic anti-tumor efficacy when AFM13 is combined with anti-PD-1 checkpoint blockade in HL PDX models, mediated by tumor-infiltrating lymphocytes, macrophages and dendritic cells, and provide strong evidence for cross-talk between innate and adaptive immunity induced by AFM13-recruited human NK-cells. Citation Format: Xing Zhao, Narendiran Rajasekaran, Uwe Reusch, Jens-Peter Marschner, Martin Treder, Holbrook E. Kohrt. Immune checkpoint inhibition by anti-PD-1 or CD137 co-stimulation enhances cytotoxicity towards CD30+ tumors mediated by the bispecific tetravalent CD30/CD16A TandAb AFM13. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2323.

Abstract 5206: Recent incidence trends of lymphoid malignancies in Hong Kong, 2001-2010

Abstract Incidence trends of lymphoid malignancies in the United States (U.S.) have been well-characterized based on analyses of data from the Surveillance, Epidemiology, and End Results Program. These analyses showed marked increases in incidence rates of non-Hodgkin lymphoma (NHL) in the early to mid-1990s, with some evidence that rates have begun to level off in the recent decade. In contrast, there are limited published data on trends in rates of lymphoid malignancies in the Chinese population, particularly for subtypes. Such analyses may provide insight into geographic differences in the descriptive epidemiological patterns of these neoplasms as well as etiological hypotheses. To explore recent trends in incidence rates in HK, yearly age-adjusted incidence rates and annual percent changes (APCs) were calculated using available data over the period 2001-2010 for overall lymphoid malignancies, overall B-cell neoplasms not including Hodgkin lymphoma (HL), overall NK or T-cell tumors, HL, plasma cell neoplasms (PCN), and specific NHL B-cell subtypes that were most common in the HK population over this time period, including diffuse large B-cell lymphoma (DLBCL), marginal zone lymphoma (MZL), and follicular lymphoma (FL). The incidence of overall lymphoid malignancies showed a steady increase in HK over the period 2001-2010 with an APC of +1.4% (95% CI = + 0.4%, + 2.3%). A steady rate of increase was further observed over the ten year period for overall B-cell neoplasms not including HL (APC = +1.6%, 95% CI = + 0.4%, + 2.9%), whereas the incidence of overall T- and NK/T cell neoplasms showed no significant fluctuation over the ten year period (APC = -1.1%, 95% CI = -3.5%, +1.5%). For the most common subtypes diagnosed in HK during the study period, an increasing trend of HL was observed over the years 2004-2010 only (APC = + 10.5%, 95% CI = + 4.9%, + 16.4%) based on the best-fitting model that included one joinpoint, and stable but non-significant rates of increase for DLBCL (APC = + 1.5%, 95% CI = - 0.7%, + 3.6%), PCN (APC = + 1.2%, 95% CI = -0.2%, + 2.6%), and FL (APC = + 2.7%, 95% CI -0.5%, 6.0%) were also observed, all over the period 2001-2010. Conversely, incidence rates of MZL showed no significant fluctuation over the study period (APC = -1.0%, 95% CI = -2.9%, +0.8%). The overall trends observed during even this relatively short time-period suggest that the rate of several lymphoid malignancies in HK may be beginning to shift in the direction of rates that were observed among individuals living in the U.S. Additional analyses of population-based registry data from other regions in East Asia, especially over longer time-periods, are needed to replicate and extend these findings. Further, our results support the need for etiological studies of lymphoid malignancies in this population, particularly as it relates to environmental, occupational and lifestyle risk factors that might influence disease rates and the genetic risk factors that potentially interact with them. Citation Format: Bryan A. Bassig, Wing-Yan Au, Oscar Mang, Roger Ngan, Lindsay M. Morton, Dennis K.M. Ip, Wei Hu, Tongzhang Zheng, Wei Jie Seow, Jun Xu, Nathaniel Rothman, Qing Lan. Recent incidence trends of lymphoid malignancies in Hong Kong, 2001-2010. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5206.

Anti-PD-1 to enhance NK-cell cytotoxicity towards CD30+ Hodgkin lymphoma induced by CD30/CD16A TandAb AFM13.

e14012Background: AFM13 is an NK-cell engaging TandAb in Ph. 2 development in Hodgkin Lymphoma (HL) and CD30+ malignancies. As immune checkpoint modulators have demonstrated clinical efficacy in a various cancers we were prompted to investigate the combination of AFM13 with several immunomodulatory antibodies. In previous experiments, we demonstrated higher efficacy of AFM13 than such antibodies and strong synergy between AFM13 and an anti-PD1 antibody in vitro, and in vivo PDX models with human HL tumors. Methods: To further investigate the immunological mechanisms we used an autologous PDX mouse model with donor-matched tumors and PBMCs from HL patients. Animals were treated with AFM13 and in combination with anti-PD1. Tumor size, tumor-infiltrating human lymphoid cells, myeloid cells and intra-tumoral cytokines were evaluated at several time-points after treatment start. Results: While monotherapy with AFM13 was reproducibly more potent than anti-PD1 in controlling tumor growth, significant synergy was...

Cyto-histopathological study of Mediastinal lesions

Background: Broad ranges of non-neoplastic and neoplastic lesions are encountered in mediastinum depending on the patient’s age and anatomical site. This study was performed to review our institutional experience of mediastinal masses and to compare the results with various other studies.Materials and Methods: This was a retrospective study conducted on 112 patients with mediastinal masses who underwent fine needle aspiration cytology and/or biopsy from 14th April 2009 to 4th March 2014 in Department of Pathology, Institute of Medicine, Tribhuvan University Teaching Hospital.Results: Out of 112 cases, 70 cases (62.5%) were benign, 38 cases (33.92%) malignant and 4 cases (3.57%) were inconclusive. Mean age of presentation was 34.94 yrs with male to female ratio of 1.2:1. Ninety-nine cases (88.4%) were seen in anterior compartment, 12 cases (10.7%) in posterior compartment and 1case (0.9%) in middle compartment. Thymoma (26 cases) was the most common lesion followed by Non Hodgkin lymphoma (17 cases) and germ cell tumor (16 cases).Conclusion: Mediastinal lesions occur more commonly in males with higher frequency of benign lesions, among which thymoma is the predominant lesion.

Social Media and the Adolescent and Young Adult (AYA) Patient with Cancer.

Over 70,000 adolescent and young adults (AYA) aged 15 to 39years are diagnosed with cancer each year in the US. The National Cancer Institute (NCI) has identified AYA cancer patients as a unique population. The most common cancers in this age group include tumors typically seen in pediatric patients such as acute lymphoblastic leukemia (ALL) and brain tumors, as well as cancers more typically seen in adult patients such as breast cancer and melanoma. In addition, some cancers have their highest incidence in AYA patients, such as Hodgkin Lymphoma, testicular cancer, and bone tumors. AYA patients face additional unique issues due to their age, not just questions about treatment choices due to lack of data but also questions about fertility, relationships, loss of autonomy, and interruptions in school/work with potentially significant financial complications. This age group also has very high rates of social media usage with up to 90% of adults aged 18 to 29 using social networking sites. In this review, we will describe the use of social media in AYAs with cancer and highlight some of the online resources for AYAs.

Epidemiology of Adolescents and Young Adults with Cancer in Europe.

To design the services for adolescents and young adults (AYAs) with cancer, we need to understand the patterns of disease and the other clinical and managerial challenges of the patient group. Cancer occurring between the ages of 15 and 39 years is 4 times less rare than cancer occurring during the first 15 years of life and consists of 2% of all invasive cancer in Europe, about 66,000 patients in Europe each year. AYAs have a unique distribution of cancer types, including the peak in incidence of Hodgkin lymphoma (HL) or germ cell tumors. The relative improvement in the survival rate in AYAs has not kept pace with that achieved in younger children, especially for acute leukemia, non-HLs, Ewing tumors and rhabdomyosarcoma. Etiological factors are under-researched and remain largely hypothetical. In this unique group of illnesses, improving AYA cancer management involves bridging interfaces. Since this has begun, outcomes have also begun to improve. The local nature of these interfaces determines the age group considered as AYA. Specific skills are necessary in the clinical, biological and psychosocial domains. Services need support from policy, clinical and administrative professionals. National policy and supranational groups such as SIOPE and ESMO are in constructive collaboration to develop this further.

Analysis and clinical correlation of infiltrating cytotoxic T cells in Hodgkin Lymphoma tumor samples using flow cytometry

Background: Progressive transformation of germinal centers (PTGC) is a disease of mostly young adults that generally presents as asymptomatic, localized or generalized lymphadenopathy. Although PTGC usually seen as nodal disease, it can occur at extranodal sites. In the differential diagnosis of chronic lymphadenopathies, infection, lymphoproliferative disorders and progressive transformation of germinal centers should be considered. The most significant entities in the differential diagnosis with PTGC are nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) and lymphocyte rich classical Hodgkin lymphoma. PTGC may precede, follow, or be concurrent with lymphomas. Objectives: To review the clinical and pathological features of PTGC, and PTGC’s relationship with lymphoid neoplasia in adult population. Material and Methods: The data of 33 patients who was diagnosed with PTCG at Hospital of Ege University School of Medicine, Pathology Department, between January 2000 and December 2014 were analyzed retrospectively. Clinical and Pathological Features: The median age of the patients was 43.8 years (26-66 years), and of 16 (48.5%) males and 17 (51.5%) females. Biopsy regions were; cervical lymph nodes (LN) 9 (27.3%), axillary LN 9 (27.3%), inguinal LN 6 (18.2%), submandibular LN 3 (9.1%), submental LN 2 (6.1%), parapancreatic LN 1 (3.0%), intraparotid LN 1 (3.0%), supraclavicular LN 1 (3.0%) and intramammarian LN 1 (3.0%). Recurrent PTGC was detected in only one (3.0%) patient. Diffuse large B cell lymphoma (DLBCL) and nodular lymphocyte predominant hodgkin lymphoma (NLPHL) was detected concurrent with PTGC in two cases. Also PTGC was detected 3 years after the diagnosis of DLBCL, NLPHL and T cell rich B cell lymphoma in 3 cases. There was no relapse in these 3 cases and no progression to lymphoma during follow-up of other patients. Conclusion: PTGC is not considered a premalignant condition but development of lymphoma have been reported rarely. If PTGC occurs in the follow of lymphoma cases, follow-ups may be intensified. NLPHL is the most important entity in differantial diagnosis of PTGC and patients should be carefully examined in terms of NLPHL.

Abstract 2469: Brentuximab vedotin-mediated immunogenic cell death

Abstract Brentuximab vedotin (ADCETRIS®) is an antibody-drug conjugate (ADC) directed against CD30. It consists of an anti-CD30 monoclonal antibody conjugated to monomethyl auristatin E (MMAE), a microtubule-disrupting agent. Brentuximab vedotin is approved for the treatment of relapsed Hodgkin lymphoma (HL) and systemic anaplastic large cell lymphoma (ALCL). Brentuximab vedotin antitumor activity is due to the binding of the ADC to CD30-expressing cells, followed by internalization, and release of MMAE after proteolytic cleavage resulting in apoptotic cell death. While Brentuximab vedotin induced cell death has been extensively studied, its potential immune modulatory activity has yet to be explored. Normal apoptosis is non-immunogenic, however multiple chemotherapeutic agents have been shown to induce a unique form of cell death termed Immunogenic Cell Death (ICD). ICD is characterized by exposure of danger-associated molecular patterns (DAMPs), many of which are Toll-like receptor ligands, which can reinitiate the immune responses suppressed by the tumor microenvironment. To test whether Brentuximab vedotin mediated tumor cell death induced ICD the phenotypic characteristics of Brentuximab vedotin-killed CD30+ HL tumor cell lines was examined. Brentuximab vedotin, dose-dependently induced surface exposure of the hallmark ICD markers calreticulin and HSP90 to similar levels induced by oxaliplatin, a chemotherapeutic agent known to mediate ICD. Since ICD is downstream of the ER stress response, Brentuximab vedotin was assessed for its ability to induce ER stress. Brentuximab vedotin treatment upregulated the apoptotic ER sensor C/EBP homologous protein (CHOP) and induced the cleavage and activation of ATF6, a transcription factor required for induction of the ER stress response. ER stress response occurred concurrently with induction of ICD markers and preceded the appearance of active caspase 3/7. These results indicate that Brentuximab vedotin mediated disruption of the microtubule network, in addition to perturbing the cell division cycle and mitosis, also induces ER stress contributing to ICD-dependent cellular toxicity. The functional consequence of ICD is activation of an immune reaction. Thus, exposure of dendritic cells to Brentuximab vedotin-killed tumor cells evoked an inflammatory phenotype including an increase in co-stimulatory markers CD86 and MHC Class II antigens, and activation of NFkB, an intermediate of inflammatory signaling pathways. Taken together, this report suggests that CD30-expressing tumor cells killed by Brentuximab vedotin may potentially activate the innate immune system to initiate antitumor immune response. It also provides a rationale for exploring therapeutic strategies that combine Brentuximab vedotin with other immune stimulatory regimens. Citation Format: Shyra J. Gardai, Angela Epp, Che-Leung Law. Brentuximab vedotin-mediated immunogenic cell death. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2469. doi:10.1158/1538-7445.AM2015-2469

Advances in the diagnosis and treatment of Hodgkin lymphoma and systemic anaplastic large cell lymphoma

Abstract Current management of Hodgkin lymphoma (HL) allows high cure rates to be achieved with initial therapy in both early-stage and advanced-stage disease. However, adverse events and the emergence of late effects are important issues for patients who respond well to frontline treatment. A further challenge in HL is the treatment of patients who are either refractory to, or relapse following, initial therapy. High-dose chemotherapy with autologous stem cell transplantation (HDCT/ASCT) is a potentially curative second-line treatment option for many patients with relapsed/refractory (R/R) HL. However, not all patients are candidates for HDCT/ASCT, and the frequency and duration of responses vary. For patients who fail HDCT/ASCT, current treatment options include further salvage chemotherapy, allogeneic transplantation or novel targeted therapies. Similar issues surround the management of R/R systemic anaplastic large cell lymphoma (sALCL), which is a rare and aggressive tumour. CD30 is strongly expressed in both HL and sALCL tumour cells and is a promising therapeutic target, as demonstrated by the activity of the novel antibody–drug conjugate brentuximab vedotin. New approaches to individualising and optimising patient management, including improved application of imaging, better prognostication through use of biomarkers and the potential use of new treatment combinations and agents targeting signalling pathways in malignant cells, may help to improve outcomes. The case studies included herein were presented at a symposium during the European Hematology Association (EHA) meeting in 2014. This publication aims to share clinical experiences and solutions to the management of HL and sALCL patients in very challenging disease settings.

How to Improve the Safe and Effective Use of Doxorubicin in Children with Cancer.

Doxorubicin, one of the anthracyclines, is a chemotherapeutic agent widely used to treat several pediatric cancers such as acute lymphocytic leukemia, acute myeloid leukemia, Hodgkin lymphoma, non-Hodgkin lymphoma, softtissue sarcoma, bone sarcoma, Ewing sarcoma, neuroblastoma, and Wilms tumor. As such, this drug has contributed substantially to the dramatic improvement of the 5-year survival rates for many of these childhood cancers [1]. However, a major potential adverse event of the use of doxorubicin is its cardiotoxicity. More than 50 % of patients will develop asymptomatic cardiac dysfunction, whereas one in six will result in clinical heart failure due to cardiomyopathy [2]. The exact causative mechanism of this devastating adverse event is, despite the fact that doxorubicin has been used for decades in many patients, not fully elucidated. There is more or less consensus about the important role of reactive oxygen species causing cardiac cell damage, progressive myocyte loss that ultimately will result in a decreased cardiac contractility [3]. To date, several clinical risk factors have been linked to the development of doxorubicin-induced cardiotoxicity and these are a higher single as well as total cumulative dose, cardiac irradiation, short infusion time duration, younger age, longer time since the treatment, and female sex [4]. In addition, much more recently, the potentially important role of genetic factors as predictors of doxorubicin-induced cardiotoxicity has been brought to our attention [5]. Multiple genetic variants in several genes associated with this cardiotoxicity have been identified [6]. However, until now, the currently available knowledge about both the clinical as well as the genetic risk factors has never resulted in sufficient discriminatory power to divide patients into groups with a high or low risk of developing cardiotoxicity. In this issue of the journal, Voller and colleagues have described a population pharmacokinetic model of doxorubicin in children in which they have used a power function to describe the relation between doxorubicin clearance and age [7]. They were able to show, for the first time, that the pharmacokinetics (PK) of doxorubicin in infants and children with cancer are age dependent. Clearly, this finding might eventually assist in designing a more tailored, safe, and effective dosing regimen for the use of doxorubicin in children with cancer. However, before this demonstrated lower clearance of doxorubicin in infants and children younger than 3 years of age will be able to change the currently used dosing regimens in pediatric patients with cancer, there is an absolute need to not only consider developmental PK but also, and probably more importantly, developmental pharmacodynamics (PD). At first glance, based on the demonstrated lower total body clearance, it seems that we need less doxorubicin in this group of young patients to reach the same amount of doxorubicin exposure as compared with older children and adolescents. However, what do we really know about the dose-concentration-effect relationship in children with cancer between 1 and 18 years of age? In other words, there is an urgent need to better This commentary refers to the article available at doi:10.1007/s40262-015-0272-4.

VI. FDG-PET as a biomarker in lymphoma: from qualitative to quantitative analysis.

VI. FDG-PET as a biomarker in lymphoma: from qualitative to quantitative analysis.

K1 and K15 of Kaposi's Sarcoma-Associated Herpesvirus Are Partial Functional Homologues of Latent Membrane Protein 2A of Epstein-Barr Virus.

The human herpesviruses Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV) are associated with Hodgkin's lymphoma (HL) and Primary effusion lymphomas (PEL), respectively, which are B cell malignancies that originate from germinal center B cells. PEL cells but also a quarter of EBV-positive HL tumor cells do not express the genuine B cell receptor (BCR), a situation incompatible with survival of normal B cells. EBV encodes LMP2A, one of EBV's viral latent membrane proteins, which likely replaces the BCR's survival signaling in HL. Whether KSHV encodes a viral BCR mimic that contributes to oncogenesis is not known because an experimental model of KSHV-mediated B cell transformation is lacking. We addressed this uncertainty with mutant EBVs encoding the KSHV genes K1 or K15 in lieu of LMP2A and infected primary BCR-negative (BCR(-)) human B cells with them. We confirmed that the survival of BCR(-) B cells and their proliferation depended on an active LMP2A signal. Like LMP2A, the expression of K1 and K15 led to the survival of BCR(-) B cells prone to apoptosis, supported their proliferation, and regulated a similar set of cellular target genes. K1 and K15 encoded proteins appear to have noncomplementing, redundant functions in this model, but our findings suggest that both KSHV proteins can replace LMP2A's key activities contributing to the survival, activation and proliferation of BCR(-) PEL cells in vivo. Several herpesviruses encode oncogenes that are receptor-like proteins. Often, they are constitutively active providing important functions to the latently infected cells. LMP2A of Epstein-Barr virus (EBV) is such a receptor that mimics an activated B cell receptor, BCR. K1 and K15, related receptors of Kaposi's sarcoma-associated herpesvirus (KSHV) expressed in virus-associated tumors, have less obvious functions. We found in infection experiments that both viral receptors of KSHV can replace LMP2A and deliver functions similar to the endogenous BCR. K1, K15, and LMP2A also control the expression of a related set of cellular genes in primary human B cells, the target cells of EBV and KSHV. The observed phenotypes, as well as the known characteristics of these genes, argue for their contributions to cellular survival, B cell activation, and proliferation. Our findings provide one possible explanation for the tumorigenicity of KSHV, which poses a severe problem in immunocompromised patients.

Downregulation of CD30, Resistance to MMAE, and Upregulation of MDR1 Are All Associated with Resistance to Brentuximab Vedotin

Background: Both Hodgkin lymphoma (HL) and anaplastic large cell lymphoma (ALCL) express surface CD30. Brentuximab vedotin (BV) is an antibody-drug conjugate that delivers a potent cytotoxic agent, monomethyl auristatin E (MMAE), specifically to cells expressing surface CD30. Although BV elicits a high response rate (75% in HL and 86% in ALCL), the majority of patients who do not attain complete response (CR) will eventually develop resistance to BV. It is not known whether resistance to BV is through a) CD 30 alterations b) resistance to cytotoxic agent MMAE or c) overexpression of drug exporters. We developed 2 BV-resistant cell models and obtained primary lymphoma samples from patients with relapsed/progressive disease post BV therapy. We examined CD30 expression, MMAE resistance, drug exporter expression, and gene expression profiles in vitro and in vivo to determine mechanisms of resistance to BV.Methods: HL cell line(L428) and ALCL cell line (KARPAS 299) were used for in vitro experiments. The selection of BV resistant cell model (L428R and KARPAS 299R) used two different approaches (pulsatile or constant exposure). Both BV resistance and MMAE resistance were confirmed by MTS assays. CD30 expression was measured by flow cytometry,qRT-PCR, and Western blotting. Drug exporter expression was measured using qRT-PCR to MDR1, MRP1, and MRP3. In vivo experiments utilized primary tumor samples from 15 HL and 4 ALCL patients who had developed relapsed/progressive disease post BV treatment. CD30 expression was assessed by immunohistocytochemistry (IHC). Gene expression profiling was performed in both parental and resistant HL and ALCL cells, and in 4 ALCL primary tumor samples using Affymetrix whole genome GeneChip® Human Genome U133 2.0 Plus.Results: MTS assay showed the IC50 of KARPAS 299R to BV shifted from 24 +/- 10 ng/ml to 28 +/- 9 ug/ml, an 1183-fold increase. MTS assay also showed the IC50 of KARPAS 299R to MMAE only increased 2-fold when compared to KARPAS 299. Flow cytometry showed downregulation of surface CD30 expression in KARPAS 299R as compared to KARPAS 299 parental (59% vs. 96%, median intensity 78 +/- 17 vs. 591 +/- 51). This downregulation was confirmed by qRT-PCR and Western blotting for CD30. As KARPAS 299R is a mixed cell population, we sorted them into CD30+ and CD30- subpopulations. We then analyzed for BV sensitivity based on CD 30 expression status in KARPAS 299R. MTS assay showed that KARPAS 299R CD30+ cells were equally as resistant to BV as KARPAS 299R CD30- cells (figure 1A). IHC performed in 4 ALCL primary tumor samples showed persistent CD 30 expression in relapsed/progressive tumor specimens post BV treatment.Gene expression profiling on KARPAS 299R showed downregulation of CD30 as compared to KARPAS 299. Gene expression profiling on pre- and post-treatment ALCL samples (8) did not show significant differences in CD30 expression. The top four upregulated genes in relapsed/progressive samples as compared to pretreatment samples were LCE3D, WNT3, TNNT, CITED2. The top four downregulated genes in relapsed/progressive samples as compared to pretreatment samples were CXCL13, C4orf7, MS4A1, and IGJ.MTS assay showed that the IC50 of L428R to BV has shifted from 32 +/- 11 ug/ml to 391 +/- 92 ug/ml, a 12-fold increase. MTS assays showed the IC50 of L428R to MMAE has increased 99-folds when compared to L428 (figure 1B). No difference was seen in CD 30 expression by flow cytometry, qRT-PCR, or western blotting between L428R vs. L428. IHC performed in 15 HL primary tumors show persistent CD30 expression in relapsed/progressive tumor specimen post-BV treatment. qRT-PCR showed upregulation of MDR1mRNA in L428R as compared to L428. Gene expression profiling on L428R showed upregulation of MDR1 as compared to L428.Conclusion: Downregulation of CD30 is seen in BV-resistant ALCL cell model. However, sensitivity to BV did not depend solely on the level of CD30 expression as CD30+ cell subpopulations still exhibited resistance to BV in vitro. Upregulation of MDR-1 and resistance to MMAE were seen in BV-resistant HL cells, rather than downregluation of CD30. Downregulation of CD30 was not seen in HL or ALCL primary tumors. Further work is ongoing to explore/validate potential targets derived from gene expression profiling in ALCL primary tumors. [Display omitted] [Display omitted] DisclosuresChen:Seattle Genetics, Inc.: Consultancy, Research Funding, Speakers Bureau, Travel expenses Other.

The classical Hodgkin lymphoma tumor microenvironment: macrophages and gene expression-based modeling.

Despite the high cure rate in classical Hodgkin lymphoma (CHL), more accurate tailoring of upfront treatment is required to maximize cure while avoiding unnecessary short- and long-term treatment side effects. To this end, the unique tumor microenvironment of CHL has been searched extensively for prognostic biomarkers. Beyond targeted immunohistochemistry (IHC) studies, gene expression profiling (GEP) of diagnostic whole tissue biopsies has allowed a de novo approach to biomarker discovery. Among numerous candidate biomarkers, an association between the number of tumor-associated macrophages in the microenvironment and outcomes after ABVD (doxorubicin + bleomycin + vinblastine + dacarbazine) chemotherapy emerged, and multiple subsequent studies have validated this biological relationship using IHC. These studies have also defined key aspects for macrophage interrogation, including the characteristics of the CD68 and CD163 antibodies, appropriate scoring methodologies, and the identification of specific patient populations in which macrophage IHC may not be prognostic. The GEP studies also led to the development of gene expression-based prognostic models for advanced-stage CHL, with new technologies allowing reliable gene expression quantitation using RNA from routinely produced formalin-fixed paraffin-embedded biopsies. The bridge to predictive biomarkers that can be used reliably to inform upfront treatment selection requires further studies to demonstrate that these biomarkers can identify robustly, at diagnosis, patients at high risk of treatment failure with ABVD and that this risk may be overcome using alternative treatments.

Pulmonary function abnormalities in childhood cancer survivors treated with bleomycin.

Bleomycin is associated with pulmonary toxic side effects including pneumonitis and pulmonary fibrosis. We evaluated the prevalence of long-term pulmonary function abnormalities in children receiving bleomycin therapy in the context of current chemotherapeutic regimens. A retrospective review of patients who received bleomycin between January 1999 and December 2011 was conducted. Abnormalities in the most recent pulmonary function test (PFT) at least 1 year after diagnosis were analyzed. Two-hundred and seven patients had received bleomycin. The results of PFT performed at least 1 year from diagnosis were available for 80 patients. Median time of follow up was 3.9 years (range 1.1-11.76 years). Median cumulative dose of bleomycin was 65 IU/m(2) (range 10-120). The most common diagnoses were Hodgkin lymphoma and germ cell tumor. At least one pulmonary function abnormality was present in 42 (52.5%) patients. When classified in groups, 22.5% patients had obstructive lung disease, 7.5% had restrictive lung disease, 28.8% had hyperinflation and 14% of patients had non-uniform distribution of ventilation. Non-Hispanic patients (OR 2.81) and children younger than 8 years (OR 4.14) had higher odds of having an abnormal PFT parameter. Very few patients had pulmonary symptoms. More than half the patients who received bleomycin had subclinical pulmonary dysfunction as evidenced by abnormalities in pulmonary function tests, although the incidence of clinical symptoms was low.

Fdg-pet in two cases of neurofibromatosis type 1 and atypical malignancies.

Patients with neurofibromatosis type 1 (nf1) are at increased risk for both benign and malignant tumours, and distinguishing the malignant potential of an individual tumour is a common clinical problem in these patients. Here, we review two cases of uncommon malignancies (Hodgkin lymphoma and mediastinal germ-cell tumour) in patients with nf1. Although (18)F-fluorodeoxyglucose positron-emission tomography (fdg-pet) has been used to differentiate benign neurofibromas from malignant peripheral nerve sheath tumours, fdg-pet characteristics for more rare tumours have been poorly described in children with nf1. Here, we report the role of pet imaging in clinical decision-making in each case. In nf1, fdg-pet might be useful in the clinical management of unusual tumour presentations and might help to provide information about the malignant potential of uncommon tumours.