T-cell Lymphoma Patients Research Articles

Bio-P-13 - Boost of innate immunity cytokines as biomarkers of response to extracorporeal photopheresis in leukemic cutaneous T-cell lymphoma patients

Extracorporeal photopheresis (ECP) emerged as a systemic first-line therapy in leukemic cutaneous T-cell lymphoma (L-CTCL) and is now being implemented in a variety of T-cell-mediated diseases. Although ECP has been used for nearly 30 years, the mechanisms of its efficacy are not sufficiently understood and biomarkers for response are lacking. We aim to investigate the immunomodulatory effects of ECP on cytokine secretion patterns in patients with L-CTCL. A total of 22 L-CTCL patients and 15 healthy donors (HDs) were retrospectively enrolled in this study. Concentrations of 16 cytokines were simultaneously quantified by using multiplex bead-based immunoassays. First, the result revealed distinct cytokine patterns between L-CTCLs and HDs. We observed significant loss of TNFa and IL-8, and significant increase of IL-9 and IL-13 in serums of L-CTCL patients compare to HDs. As a next step, L-CTCL patients who received ECP were classified to responder and non-responder to treatment according to their quantitative reduction of malignant burden in the blood. We evaluated cytokine levels in culture supernatants from patient peripheral blood mononuclear cells (PBMCs) at baseline and over 27 weeks after ECP. Strikingly, PBMCs purified from ECP responders released statistically higher concentration of innate immunity cytokines such as IL-1a, IL-1b, GM-CSF and TNFα in comparison with ECP non-responders. In parallel, clinical observations documented complete clearance of erythema, reduction of malignant clonal T cells in the blood, and a potent boost of relevant innate immunity cytokines in individual L-CTCL patients. To conclude, our results demonstrated that ECP stimulates innate immune network, and this implied how ECP facilitates redirection of tumor-biased immunosuppressive microenvironment toward active anti-tumor responses. The alteration of TNFα, IL-1a, IL-1b and GM-CSF can possibly be used as biomarkers of response to ECP in leukemic CTCL patients.

Disparity analysis and prognostic value of pretreatment whole blood Epstein-Barr virus DNA load and Epstein-Barr encoding region status in lymphomas: A retrospective multicenter study in Huaihai Lymphoma Working Group.

Elevated Epstein-Barr virus (EBV) DNA load is common in lymphomas. However, it remains unclear whether the disparity in viral load and its prognostic value in lymphomas are correlated with Epstein-Barr encoding region (EBER) status. In this retrospective multicenter study, we collected the data of pretreatment whole blood EBV DNA (pre-EBV DNA) and EBER status and evaluated their disparity and prognostic values in lymphomas. A total of 454 lymphoma patients from December 2014 to August 2020 were retrospectively retrieved. Mann-Whitney Utest, Kruskal-Wallis test and Bonferroni's adjustment were used to explore the disparity of EBV DNA and EBER status in lymphomas. Time-dependent receiver operating characteristic analysis and MaxStat analysis were used to determine optimal cutoff points of pre-EBV DNA load. Univariable and multivariable Cox proportional hazards models were established for the estimation of prognostic factors. The positive rate of EBV DNA in natural killer T-cell lymphoma (NKTL) patients was higher than that in diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL) and Hodgkin lymphoma (HL) patients, and the median positive pre-EBV copy number of NKTL was also higher than that of FL and DLBCL. EBV DNA could clearly distinguish the prognosis of DLBCL, NKTL, HL and peripheral T-cell lymphoma, and the integration of EBER status and EBV DNA could differentiate the prognosis of HL patients. Multivariable results revealed that pre-EBV DNA load had an effect on the prognosis of NKTL, FL and DLBCL. The status of pre-EBV DNA and EBER were disparate. Whole blood pre-EBV DNA predicted the prognosis of lymphomas, and the combination of EBV and EBER status could differentiate the prognosis of HL.

Antagonism of inhibitors of apoptosis proteins reveals a novel, immune response-based therapeutic approach for T-cell lymphoma

Tolinapant (ASTX660) is a potent, nonpeptidomimetic antagonist of cellular inhibitor of apoptosis proteins 1 and 2 (cIAP1/2) and X-linked IAP, which is currently being evaluated in a phase 2 study in T-cell lymphoma (TCL) patients. Tolinapant has demonstrated evidence of single-agent clinical activity in relapsed/refractory peripheral TCL and cutaneous TCL. To investigate the mechanism of action underlying the single-agent activity observed in the clinic, we have used a comprehensive translational approach integrating in vitro and in vivo models of TCL confirmed by data from human tumor biopsies. Here, we show that tolinapant acts as an efficacious immunomodulatory molecule capable of inducing complete tumor regression in a syngeneic model of TCL exclusively in the presence of an intact immune system. These findings were confirmed in samples from our ongoing clinical study showing that tolinapant treatment can induce changes in gene expression and cytokine profile consistent with immune modulation. Mechanistically, we show that tolinapant can activate both the adaptive and the innate arms of the immune system through the induction of immunogenic forms of cell death. In summary, we describe a novel role for IAP antagonists as immunomodulatory molecules capable of promoting a robust antitumor immune response in TCL.

TCL-461: Clinical Activity of Systemic VSV-IFNβ-NIS Oncolytic Virotherapy in Patients with Relapsed Refractory Hematologic Malignancies

<h3>Context</h3> This is the first in human Phase 1 dose escalation trial of oncolytic Vesicular Stomatitis Virus (VSV) incorporating interferon beta and sodium iodine symporter transgenes (VSV-IFNβ-NIS) to be administered intravenously in patients with relapsed and refractory hematologic malignancies. <h3>Objective</h3> The primary objective was to determine the maximum tolerated dose of VSV-IFNβ-NIS as a single agent. The secondary objectives were to determine safety and efficacy of VSV-IFNβ-NIS. Correlative objectives included monitoring viremia and virus shedding. <h3>Design</h3> This was a 3+3 phase 1 clinical trial from April 2017 to August 2020 including adult patients (pts) with relapsed refractory multiple myeloma (MM), T-cell lymphoma (TCL), or acute myeloid leukemia (AML). <h3>Results</h3> Fifteen patients received VSV-IFNβ-NIS: MM (7), TCL (7), and AML (1); median age was 64 years, 60% male, and median prior lines of systemic cancer therapies was 5 (range, 2 to 11). Three patients were treated at each dose level (DL) 1 through 3 (respectively 0.05, 0.17, and 0.5 x 10¹¹ TCID₅₀), and 6 at DL 4 (1.7x10¹¹ TCID₅₀). There were no dose-limiting toxicities. Hematologic AEs, particularly lymphopenia (73%) were the predominant grade 3 and 4 AEs. Non-hematologic AE of interest were grade 1 (6.7%) and 2 (46.6%) cytokine release syndrome, of which only 1 patient required transient norepinephrine support. Objective responses were seen in patients with TCL. There was a 3-month partial response (PR) in a patient treated at DL 2 after 11 prior systemic therapies. However, compelling responses were seen at DL 4 with a deepening 6-month PR in a patient with PTCL and ongoing CR, even 14 months after VSV in another patient with cutaneous relapse of PTCL; both patients received 5 prior lines of therapy. Viremia rapidly declined after infusion, and there was no significant viral shedding. Neutralizing anti-VSV antibodies developed by day 29. IFNβ appeared to indicate intratumoral amplification with TCL patients mounting the highest levels. <h3>Conclusions</h3> A single intravenous dose of VSV-IFNβ-NIS up to 1.7x10¹¹ is safe in heavily pre-treated patients with hematologic malignancies and appears to be most effective as a single agent at DL 4, especially in patients with TCL. Future trials of combination strategies with immune-modulatory drugs are currently being planned. <h3>Grant Acknowledgements</h3> Mayo Clinic Multiple Myeloma Specialized Program of Research Excellence (SPORE), Grohne gifts, Phillips gift, and Liu gift.

Second neoplasm in cutaneous T-cell lymphoma patients: a marker of worse prognosis?

Background Epidemiologic studies have shown that cutaneous T-cell lymphoma (CTCL) patients have an increased risk of the development of a second neoplasm (SN). The aim of our study was to evaluate the risk of SN and to correlate any possible change in CTCL course after the diagnosis of a subsequent neoplasm. Methods A ten-year retrospective study was carried out in two centres (Bologna and Florence) all the patients who developed a SN six months at least after a CTCL were included. Two groups were selected: Group 1 featuring patients who developed a SN and Group 2 characterised by patients affected by MF age and sex-matched with Group 1 (control group). Data concerning any stage change after SN, time between MF and SN onset, modified Severity Weighted Assessment Tool (mSWAT) score before and after SN, concerning Group 1 and after a median time of 36 months in Group 2 were analysed. Results Thirteen patients were detected. Before SN onset, early MF patients were mainly present, while SN cases in advanced-stage (ten patients) were observed. SN type predominant was lung cancer, along with prostate and pancreatic cancer, while isolated cases presenting with vulvar, colon, mammalian, prostate cancer along with Hodgkin's Lymphoma. Mean mSWAT at MF diagnosis and after SN showed a significant difference (p value=0.0037). After SN diagnosis, nine patients experienced an MF stage progression and ten patient died at follow up. Conclusions In all the instances, statistical analysis showed that mean mSWAT score before/after SN diagnosis had a significantly difference (p value=0.0037) suggesting that patients with a SN may have a worse clinical outcome. By secreting immunosuppressive cytokines or recruiting immunosuppressive cells, a sort of mutual help between the two neoplasms may be prompted. Our data suggest that SN development in MF patients may be regarded as a worse prognostic marker.

Correlation between the \u03b3 passing rates of IMRT plans and the volumes of air cavities and bony structures in head and neck cancer

BackgroundBoth patient-specific dose recalculation and γ passing rate analysis are important for the quality assurance (QA) of intensity modulated radiotherapy (IMRT) plans. The aim of this study was to analyse the correlation between the γ passing rates and the volumes of air cavities (Vair) and bony structures (Vbone) in target volume of head and neck cancer.MethodsTwenty nasopharyngeal carcinoma and twenty nasal natural killer T-cell lymphoma patients were enrolled in this study. Nine-field sliding window IMRT plans were produced and the dose distributions were calculated by anisotropic analytical algorithm (AAA), Acuros XB algorithm (AXB) and SciMoCa based on the Monte Carlo (MC) technique. The dose distributions and γ passing rates of the targets, organs at risk, air cavities and bony structures were compared among the different algorithms.ResultsThe γ values obtained with AAA and AXB were 95.6 ± 1.9% and 96.2 ± 1.7%, respectively, with 3%/2 mm criteria (p > 0.05). There were significant differences (p < 0.05) in the γ values between AAA and AXB in the air cavities (86.6 ± 9.4% vs. 98.0 ± 1.7%) and bony structures (82.7 ± 13.5% vs. 99.0 ± 1.7%). Using AAA, the γ values were proportional to the natural logarithm of Vair (R2 = 0.674) and inversely proportional to the natural logarithm of Vbone (R2 = 0.816). When the Vair in the targets was smaller than approximately 80 cc or the Vbone in the targets was larger than approximately 6 cc, the γ values of AAA were below 95%. Using AXB, no significant relationship was found between the γ values and Vair or Vbone.ConclusionIn clinical head and neck IMRT QA, greater attention should be paid to the effect of Vair and Vbone in the targets on the γ passing rates when using different dose calculation algorithms.

Analysis Of Peripheral T-Cell Lymphoma (PTCL) Subtype By Race and Geography Using The Comprehensive Oncology Measures For Peripheral T-Cell Lymphoma Treatment (COMPLETE) Dataset

Background COMPLETE is a national registry of PTCL patients designed to describe patient characteristics, treatments and outcomes in a ‘real world’ setting. Previous work has suggested that there may be geographic and/or racial differences in the incidence of PTCL subtypes. Using the COMPLETE dataset, we explored the influence of race and geography in the United States on the incidence of PTCL subtypes and examined for interactions between these two variables. Methods We identified patients with locked baseline records and performed univariate analyses comparing disease subtype, stratified by race and geography. A multivariable, multinomial response regression model was used to predict the classification of PTCL subtype based upon race and geographic region. An interaction term for race and region was assessed via a likelihood ratio test and found to be not significant (p = 0.43), thus was removed from the model. Results A total of 247 patients had locked baseline records. Given the small numbers of Asian (7 patients, 3%) and other/race unknown patients (6, 3%), only Black (43, 17%) and White (191, 77%) patients were included in the analyses. Results of the univariate analyses can be found in [Table 1][1]. PTCL-not otherwise specified (NOS) accounted for the largest histologic subtype (35% of patients), consistent with SEER (31%) and other data sources. A trend was noted in distribution of subtype by race and significant differences were seen in histologic distribution by region, with a higher proportion of angioimmunoblastic T-cell lymphoma (AITL) patients diagnosed in the West and a smaller proportion in the South. A higher proportion of patients diagnosed with PTCL-NOS were in the South compared to other regions while a higher proportion of patients in the Northeast were diagnosed with other histologic subtypes. The multinomial regression model suggests there was a relatively high likelihood of being diagnosed with PTCL-NOS for both races in the Midwest, South and West and a relatively high likelihood of being diagnosed with other subtypes of PTCL for patients in the Northeast. White patients in the West have the highest likelihood of being diagnosed with AITL. View this table: Table 1 Results of Univariate Analyses of PTCL Subtype by Race and Region Conclusions These results suggest there is a modest, independent influence of both race and region in the United States on histologic subtype of PTCL. Whether geographic differences are due to patient selection bias or potentially due to environmental factors cannot be determined in this study. As the sample size grows, further analyses are warranted to examine if the effect of race and region is enhanced or diminished while controlling for factors not measured in the current model. Disclosures: Carson: Allos: Consultancy, Speakers Bureau; Celgene: Consultancy, Speakers Bureau. Foss: merck: Research Funding; spectrum: Research Funding; eisai: Membership on an entity’s Board of Directors or advisory committees; millenium: Honoraria, Membership on an entity’s Board of Directors or advisory committees; celgene: Honoraria, Research Funding; seattle genetics: Research Funding. Pinter-Brown: Allos: Consultancy, Membership on an entity’s Board of Directors or advisory committees. Horwitz: Spectrum (Allos): Consultancy, Research Funding. Rosen: Allos: Consultancy, Honoraria. Gisselbrecht: Allos: Consultancy, Membership on an entity’s Board of Directors or advisory committees. Hsi: Allos: Research Funding; Seattle Genetics: Speakers Bureau; Eli Lilly: Research Funding; Abbott: Research Funding; Cellerant Therapeutics: Research Funding; BD Biosciences: Research Funding; Millennium: Research Funding. [1]: #T1

Correlation between the prevalence of T-cell lymphomas and alcohol consumption.

Alcohol is a psychoactive substance that causes dependence, with many thousands of years in the history of mankind, being widely used in different cultures. According to the International Agency for Research on Cancer, alcohol is involved in the development of cancer, being directly associated with it. Considering that alcohol is involved in the initiation and dissemination of gastrointestinal malignancies, the objective of the study was to assess its role in the pathogenesis of T-cell lymphomas, as well as its possible correlation with chronic consumption. The patient cohort was compiled from the Sixth Medical Center of the People's Liberation Army Navy General Hospital in Beijing, China. A total of 30 patients matched the criteria and were enrolled in the study. Statistical analysis of the raw data was performed using R Statistics version R 3.5.1. released on the 29.08.2018. Our data demonstrate that the most common extranodal involvment of T-cell lymphoma patients is represented in decreasing order by bone marrow, peritoneum, rhino-oropharynx and the liver-biliary system. Nodal involvement is mainly represented in decreasing order by the laterocervical, axillary, mediastinal and inguinal regions. These findings may be of value in further research and practical/clinical settings. Fever is the most common clinical feature and was present in most studied patients.

Outcome of Patients with Gamma Delta T Cell Lymphomas: Analysis of 1429 Cases Registered in the Prospective T-Cell Project

Introduction: Current treatment strategies for mature or peripheral T-cell lymphomas (PTCL) patients (pts) are largely unsatisfactory and outcomes for patients (pts) with rare gamma delta subtypes have not been well described. The purpose of the present analysis was to analyze clinical features and treatment outcomes for 99 pts with gamma delta subtypes (hepatosplenic T cell lymphoma [HSTCL], enteropathy T cell lymphoma [EATL ], and peripheral gamma delta T cell lymphoma [PGDT]) retrieved from an international prospective T cell lymphoma registry, the T-cell Project (TCP).Methods: In this study, data were extracted from 1429 cases of newly diagnosed PTCL registered by 74 institutions world-wide in the T-cell Project and on whom baseline data, information on first line treatment, response to initial therapy, time to relapse and salvage treatment were available. For this analysis, 99 pts were identified who met the criteria of gamma delta subtype, including 24 with HSTCL, 62 with EATL, and 13 with PGDT. Central pathology review confirmed the diagnosis in all cases. Data for prognostic factors, initial and salvage therapies, and outcomes (Overall Survival [OS] and Progression-free Survival [PFS]) were extracted.Results: Of the 99 cases identified, geographic distribution varied. Of EATL, 39 were from Europe, while HSTCL was slightly more frequent in the USA (44%) vs. Europe (16%). PGDT was found in USA and Europe, with no cases in South America and 2 cases in Asia. The median age for the 99 cases was 55 (range 18-86). The majority of patients were under age 60; prognostic factors included B symptoms (59%), advanced disease (69%), and monocyte count <800.mm3 (69%). Treatment records were locked and analysis was performed on 74 patients. Of those, front line therapy chosen by the treating physician consisted of chemotherapy alone (CT) in 53 (71%), CT followed by high dose therapy/stem cell rescue (HDT) in 9 (12%), and supportive care in 11. Anthracycline based CT was used in 56 (88%) of cases, but 6 (46%) of HSTCL patients received a non-anthracycline regimen. The overall response rate (ORR) was 45% with 20 (31%) CR (14 in CT alone patients and 6 in HDT patients). CR occurred in 40% of HSTCL, 30% of EATL, and 25% of PGDT. Forty-two percent of patients were refractory to front line therapy. Eleven pts underwent HDT in the salvage setting; of those only 3 pts with EATL had a CR, while one with EATL and one with PGDT had PR. At a median follow up of 51 months (95% CI 36-66), 47% of pts were alive, with a median OS of 13 months and median PFS of 11 months. There is no difference in OS or PFS at 3 or 5 yrs between EATL, HSCTL and PGDT (P=0.24 for OS, P=0.682 for PFS).Conclusion: Because the gamma delta T cell lymphomas are rare and are under-represented in clinical trials, there is little data available describing treatment approaches and outcomes. In this cohort of 99 pts from the TCP registry, outcomes using the physician guided regimens are described and demonstrate that anthracycline based CT was the most common choice in first line therapy for EATL and PGDT, while non anthracycline based therapy is chosen for HSTCL. Overall CR rates were lower than those seen for nodal PTCL subtypes using similar treatment approaches. Surprisingly, despite NCCN guidelines suggesting that HDT is considered for consolidation in first remission, only a proportion of patients could undergo HDT, and HDT in the salvage setting had limited success. These results suggest that the incorporation of novel agents in the front line and the implementation of consolidation strategies, including stem cell transplantation and maintenance therapies should be considered for gamma delta T cell lymphoma subtypes. [Display omitted] DisclosuresFoss:celgene: Honoraria; immune design: Research Funding; spectrum: Honoraria, Speakers Bureau; Eisai: Honoraria; seattle genetics: Speakers Bureau. Horwitz:Infinity/Verastem: Consultancy, Research Funding; Mundipharma: Consultancy; ADCT Therapeutics: Research Funding; Kyowa-Hakka-Kirin: Consultancy, Research Funding; Seattle Genetics: Consultancy, Research Funding; Forty-Seven: Consultancy, Research Funding; HUYA: Consultancy; BMS: Consultancy; Aileron Therapeutics: Research Funding; Celgene: Consultancy, Research Funding; Millenium/Takeda: Consultancy, Research Funding. Kim:Mundipharma: Research Funding; Celltrion, Inc: Consultancy, Honoraria; Donga: Research Funding; J&J: Research Funding; Kyowa-Kirin: Research Funding; Takeda: Research Funding; Novartis: Research Funding; Roche: Research Funding. Hitz:Celgene: Consultancy. Gutierrez:JANSSEN: Consultancy, Research Funding, Speakers Bureau; ROCHE: Research Funding, Speakers Bureau; SERVIER: Speakers Bureau; GILEAD: Honoraria; TAKEDA: Speakers Bureau; PFIZER: Consultancy. Davies:Roche: Consultancy, Honoraria, Research Funding; Celgene: Honoraria, Other: Advisory Board, travel expenses to attend conference, Research Funding; Gilead: Consultancy, Honoraria, Other: Travel expenses to attend conference, Research Funding; Takeda: Consultancy, Honoraria, Other: Travel expenses to attend conference, Research Funding; Bayer: Research Funding; Acerta Phamra: Research Funding; Karyopharma: Honoraria, Research Funding; Pfizer: Honoraria, Research Funding; Janssen: Honoraria, Research Funding; GSK: Research Funding; CTI: Consultancy, Honoraria, Other: Travel expenses to attend conference; Seattle Genetics: Research Funding. Federico:takeda: Honoraria, Research Funding.

The Role of Upfront Hematopoietic Stem Cell Transplantation (HSCT) in Peripheral T-Cell Lymphoma (PTCL) Patients in Complete Remission (CR) with a Special Focus on Nodal PTCL: Report from the Comprehensive Oncology Measures for Peripheral T-Cell Lymphoma Treatment (COMPLETE), a Prospective Multicenter Cohort Study

The Role of Upfront Hematopoietic Stem Cell Transplantation (HSCT) in Peripheral T-Cell Lymphoma (PTCL) Patients in Complete Remission (CR) with a Special Focus on Nodal PTCL: Report from the Comprehensive Oncology Measures for Peripheral T-Cell Lymphoma Treatment (COMPLETE), a Prospective Multicenter Cohort Study

Generation and Application of T-Cell Lymphoma Patient Derived Tumor Xenograft Models

Introduction:Although pivotal “driver mutations” have been described in T-cell lymphomas (T-NHL), targetable dependences are lacking for many entities. As result, T-NHL patients experience dismal therapeutic responses and have a poor survival. Therapeutic improvements have been hindered by the lack of informative models. Patient-Derived-Tumor-Xenografts (PDTX), recapitulating many of the biological features of primary cancers, represent a novel and powerful platform. Here, we constructed a library of T-NHL PDTX to foster drug discovery programs and new patient-tailored approaches.Methods:Fresh and/or viable cryopreserved samples from primary T-NHL were obtained from Turin, WCM, and MSKCC hospitals. For the initial tumorgraft implants, lymphoma fragments, PBMC or bone marrow cells were implanted in NSG-B2m mice. Seeds from arising tumors were then serially transplanted. MRI was used to follow tumor growth and dissemination. The lymphoma phenotype was assessed by multi-channel flow cytometry and immunohistochemistry. We used TCR gene rearrangement analyses, whole exome sequencing (WES), total RNAseq, and CHIP-Seq (STAT3, methylation marks) to annotate primary and PDTX samples. For the in vitro studies, PDTX were digested and stromal cells were excluded by panning. Enriched Patient-Derived Tumor Cells (PDTC) were cultured in either RPMI/20% FBS or StemSpan/10% KSR media. High-throughput drug screening was performed using a library of 432 compounds (1 µM for 72hrs). Cell titer glo/blue, trypan blue cell count, and annexin-V/7AAD were used to assess cell viability, apoptosis, and proliferation.Results:We implanted 123 fresh T-NHL samples, resulting in the generation of 41 PDTX lines (Fig.1). In addition, 14 implants led to the growth of lymphoblastoid EBV+ populations. All PDTX lines were successfully propagated (5-26 serial passages) with the exception of angioimmunoblastic T-cell lymphoma (AITL) PDTX, which displayed a delay in engraftment over time, and eventually ceased growing (passage 3-7). Serially propagated PDTX showed distinct immunophenotypic profiles, which closely reflected their corresponding primary samples. TCR gene rearrangement studies confirmed the faithful correspondence between donor and PDTX over different generations. WES and total RNA-Seq on primary and serially propagated PDTX (at least T1-T5) yielded a rich landscape of mutations shared by primary tumors and PDTX derivatives. On going mutations were also defined along propagations. CHIP-Seq assay defined a STAT3 classifier in STAT+ PDTC.In vitro, all PDTC were shortly maintained (<7days), with some surviving for 6-8 weeks. To overcome their limited growth in vitro, we tested whether stromal cells (i.e. E4-Endothelial cells, E4-EC) could enhance their survival and/or growth. Indeed, co-culturing of PDTC with E4-ECs rescued cell death induced by ex vivo long-term propagation (peripheral T-cell lymphoma not otherwise specified and NK/T-Cell Lymphoma, PTCL-NOS and NK) or starvation (anaplastic large cell lymphoma, ALCL).PDTC were also used to perform a drug screening targeting major signaling pathways (>3x replicates of 8 ALCL, 1 NK, and 2 PTCL-NOS), demonstrating that each PDTX exhibited a unique drug sensitivity profile, mostly dictated by its genomic landscape. This was epitomized by two PDTC (IL-2 and IL-19) derived from the same PTCL-NOS patient bearing an activating V658F JAK1 somatic mutation. These two PDTC, once exposed to JAK inhibitors, displayed down-regulation of pSTAT3 (24hr) and massive cell death (T3-T6, >95% at 72hrs). Similar results were seen with ALK inhibitors in ALK+ ALCL PDTC (n=5). Moreover, specific drugs were identified to bypass refractory phenotypes in selective PDTC (i.e. proteasome and topoisomerase inhibitors).Conclusions:We built up a large and representative PDTX library from major T-NHL entities. Immunophenotyping, genomics and functional studies showed a close correspondence between primary samples and correspondent PDTX derivatives. Our in vitro models provide a starting point for elucidating stromal:tumor cell interactions that contribute to lymphoma growth and survival in vivo, and evidence that a drug screening can be successfully executed on PDTCs. We predict that this cohort may be a valuable tool for testing novel therapeutic compounds and fostering personalized therapeutic approaches in T-NHL patients. [Display omitted] DisclosuresThompson:Agios Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees, Other: Founder; Merck: Membership on an entity's Board of Directors or advisory committees; Charles River Laboratories: Membership on an entity's Board of Directors or advisory committees. Aster:Eli Lilly and Company: Membership on an entity's Board of Directors or advisory committees. Ruan:Cell Medica: Research Funding; Pharmacyclics: Research Funding; Seattle Genetics: Consultancy, Research Funding; Celgene: Consultancy, Research Funding. Dogan:Seattle Genetics: Consultancy, Membership on an entity's Board of Directors or advisory committees; Peer Review Institute: Consultancy; Celgene: Consultancy; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees; Roche Pharmaceuticals: Consultancy. Cerchietti:Celgene: Research Funding; Lymphoma Research Foundation: Research Funding; Leukemia and Lymphoma Society: Research Funding; Weill Cornell Medicine - New York Presbyterian Hospital: Employment. Horwitz:Celgene: Consultancy, Research Funding; Kyowa-Hakka-Kirin: Consultancy, Research Funding; Infinity/Verastem: Consultancy, Research Funding; Seattle Genetics: Consultancy, Research Funding; BMS: Consultancy; HUYA: Consultancy; Aileron Therapeutics: Research Funding; ADCT Therapeutics: Research Funding; Millenium/Takeda: Consultancy, Research Funding; Mundipharma: Consultancy; Forty-Seven: Consultancy, Research Funding.

Enhanced Spontaneous Anti-Tumor Cytotoxicity of Natural Killer Cells in Cutaneous T-Cell Lymphoma Patients

Cutaneous T-cell lymphoma (CTCL) is characterized by the expansion of malignant CD4+ T cells in the skin. There are two main subtypes of CTCL, mycosis fungoides (MF) and Sézary syndrome (SS). Previous studies have demonstrated defects in cell-mediated immunity, including altered cytokine profiles and decreased neutrophil function, and patients often have recurring bacterial and viral infections. Recent studies have shown increased expression of interleukin (IL)-15 in malignant CD4+ T cells in CTCL patients (Mishra et al., Cancer Discovery, 2016). Since IL-15 can enhance NK cell differentiation and activation, we hypothesized that NK cells from CTCL patients might have phenotypic and functional alterations due to these changes in homeostatic IL-15 levels.To evaluate the absolute number of NK cells in CTCL patients, CD56+Lineage- lymphocytes were quantitated by flow cytometry. SS patients had significantly fewer NK cells as compared to normal donors (mean ± SEM of absolute cell numbers in normal donors=0.2442 ± 0.02, n=51; SS=0.1072 ± 0.02, n=9; MF=0.2161 ± 0.01, n=112; unpaired t-test normal vs. SS, p=0.007). NK cell counts were significantly associated with overall survival, with the short-term risk of death increasing by 87.5% for every increase in 0.1 of absolute NK cell counts on univariate analysis utilizing the Cox proportional hazards model (Figure 1A; p=0.0413). We then examined the cytotoxic capacity of NK cells (CD56+Lineage-) freshly purified from both MF and SS patients by standard chromium release assay. There was a significantly higher level of specific lysis of tumor cells (K562 target cells) by NK cells from CTCL patients as compared to normal donors (Figure 1B, at 50:1 ratio mean ± SEM of percent NK cell cytotoxicity in normal vs CTCL = 27.47 ± 4.76, n=13 vs. 52.54 ± 4.88, n=12, unpaired t-test, p=0.001). Upon further transcript analysis of NK cell cytolytic mediators from CTCL patients compared to normal donors, we observed significantly increased levels of perforin (mean ± SEM of relative RNA in normal vs CTCL = 91.39 ± 9.18, n=3 vs. 485.7 ± 43.15, n=7, unpaired t-test, p=0.0004) and granzyme B (143 ± 23.79, n=3 vs. 431 ± 68.13, n=7 unpaired t-test, p=0.03). In addition, we observed increased levels of interferon-gamma and Fas Ligand (5.149 ± 0.4775, n=3 vs.13.93 ± 1.52, n=7; unpaired t-test p=0.007), indicating NK cells from CTCL patients exhibited consistently elevated activation patterns as compared to normal donor NK cells. We then examined the expression pattern of the IL-15 receptor complex on NK cells in CTCL patients as compared to normal donors by RNA sequencing. There was a significant elevation in IL-15Rα (mean ± SEM of relative RNA in normal vs CTCL = 0.8677 ± 0.1018, n=3 vs. 1.772 ± 0.2055, n=7; p=0.03) and IL-15Rγ (57.02 ± 7.525, n=3 vs. 154.7 ± 12.14, n=7; p=0.001), while IL-15Rβ was also elevated (128 ± 23.68, n=3 vs. 223.2 ± 25.43, n=7; p=0.056). Next we evaluated protein levels of phosphorylated signal transducer and activator of transcription 5 (p-STAT5) in fresh whole blood as the surrogate marker of IL-15 signaling. Preliminary findings demonstrate an increase in p-STAT5 in CTCL patients as compared to normal donors (mean ± SEM of fluorescent intensity in normal donors vs CTCL patients =1506 ± 90.82, n=5 vs. 1270 ± 50.59, n=4; unpaired t-test; p=0.07).These data suggest that NK cell activation is enhanced in patients with CTCL, including MF patients with disease localized to the skin. Further investigations aimed at understanding the disconnect between NK cell numbers, activation, and disease control in CTCL patients are warranted. [Display omitted] DisclosuresPorcu:Kiowa: Research Funding; Miragen: Research Funding; Celgene: Research Funding; Tetralogic: Research Funding; Innate Pharma: Research Funding; Kura: Research Funding; Galderma: Research Funding; Cell Medica: Research Funding. William:Miragen: Consultancy, Honoraria.

Mogamulizumab Monotherapy in the Treatment of Relapsed/Refractory Peripheral T-Cell Lymphoma or Cutaneous T-Cell Lymphoma Patients in Single-Institution Experience

Objectives; T-cell lymphomas generally have a poor outcome with shorter long-term survival compared with B-cell lymphomas. CC chemokine receptor 4 (CCR4) is highly expressed by aggressive peripheral T-cell lymphoma, not otherwise specified (PTCLs), particularly adult T-cell leukemia/lymphoma (ATL) and cutaneous T-cell lymphomas (CTCLs). Mogamulizumab is a humanized immunoglobulin G1 monoclonal antibody that targets CCR4. In phase II study, Mogamulizumab has been shown to high clinical efficacy and low toxicity. However, its efficacy has not been well evaluated in the clinical setting. We retrospectively analyzed the efficacy and safety of Mogamulizumab of against relapsed/refractory non- ATL T-cell lymphomas.Methods; Fourteen patients with relapsed/ refractory CCR4 positive PTCLs or CTCL were treated with weekly Mogamulizumab (1.0 mg/kg) alone at Iwate Prefectural Central Hospital between January 2015 and June 2017. CCR4 expression was determined by immunohistochemistry by a central laboratory center (SRL, Tokyo, Japan). In the first line, most patients receive commonly anthracycline-containing induction chemotherapy regimens such as CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone). The patients had received a median of one previous chemotherapy regime (range 1-3).We retrospectively evaluated (1) the best overall response, (2) progression-free survival (PFS), (3) overall survival (OS), (4) the best response according to disease site(peripheral blood; PB, lymph nodes; LN and skin) and (5) safety. Response assessments were performed after eighth mogamulizumab infusions according to the 1999 IWG criteria guidelines for response assessments for NHL (IWG-NHL). Treatment efficacy in patients with CTCL was evaluated by using a Global Response Score. Treatment was discontinued if progressive disease (PD) was evident.For safety evaluations, adverse events (AEs) were graded according to the National Cancer Institute Common Terminology Criteria for AEs, version 4.0.Results; The median age was 69 years old (range 48-85). Eight patients are male. The diagnosis comprised PTCLs (n=12) and CTCL (n=2). The median follow up periods and median number of Mogamulizumab infusion were 293 days (range 49- 681) and 8 courses (range 2- 9). (1) Complete remission (CR) was achieved in two patients (16.7%) and partial remission in one patients (8.3%), stable disease (SD) in two patients (16.7%), respectively. And (2) median PFS was 88 days and (3) median OS was 475 days. Peripheral blood, lymph node and skin lesion were affected in two, ten, and six patients, respectively. (4) In PB, complete response (CR) was obtained in all two patients (100%) and two patients were achieved CR in skin lesion (33.3%). In LN, only one of ten patient manifested CR(10.0%). This patient's tumor burden is low. (5) With regard to safety issues, hematological AEs were frequently observed, and lymphocytopenia occurred in 62.5% of patients. However, the AEs were controllable and tolerated. Non-hematological AEs, including fever, infusion reactions and skin trouble , were frequently observed (37.5%, 25.0%, and 37.5%, respectively). Grade 3/4 non-hematological AEs were scarcely observed.Conclusions; Although Mogamulizumab monotherapy is effective with highly safety, the patients who had swollen lymph node may need combination chemotherapy with mogamulizumab. The number of patients was small in this retrospective analysis, further investigation are required. DisclosuresNo relevant conflicts of interest to declare.

Antileukemic Activity and Pharmacodynamics of Intravenous Forodesine (BCX-1777), a Novel Purine Nucleoside Phosphorylase (PNP) Inhibitor, in Phase I/II Trials in Patients with Advanced T-Cell Malignancies.

PNP represents an attractive target for drug development based on the profound, selective impairment of T-cell function seen in individuals with an inherited deficiency of PNP. Forodesine, a potent, specific transition-state analog inhibitor of PNP, is currently being investigated for safety and efficacy in T-cell and other malignancies. In a phase I/II single-center, intrapatient dose-escalation study, forodesine (starting dose: 40 mg/m2) was given to 5 patients with relapsed/refractory, aggressive T-cell leukemia/ lymphoma via a 30-min IV infusion, followed 24 hrs later by doses administered every 12 hrs for a total of 9 doses; courses could be repeated every 3–4 weeks, as required. Two patients with refractory T-cell prolymphocytic leukemia (T-PLL) and two patients with T-cell acute lymphoblastic leukemia (T-ALL) showed a decrease in leukemic or prolymphocytic counts in peripheral blood and/or bone marrow. All responders had increases in plasma 2′-deoxyguanosine (dGuo) (Cmax: 2.9 to 34 μM; baseline concentration: <.004 μM) and 4/5 responders showed elevated intracellular dGTP levels (10- to 60-fold). Forodesine was generally safe and well tolerated at all dose levels with no dose-limiting toxicities and only two adverse events (ie, acute renal failure and neutropenic infection) that were possibly drug-related. These promising results led to a phase II multi-center, open-label, repeat-dose trial in patients with advanced T-cell malignancies, in which forodesine (40 mg/m2) is infused for 30 mins for 5 days followed by at least 2 non-treatment days for up to 5 additional cycles. Out of 3 patients enrolled to-date, 2 T-ALL patients completed 6 courses of therapy with a dramatic reduction in WBC count (Fig 1A and 1B); 1 splenic T-cell lymphoma patient discontinued due to disease progression. Forodesine resulted in striking elevations in plasma dGuo. Peripheral blast cells in Patient 01 were undetectable at the start of the second course and remained undetectable after 6 courses (Fig 1A), and the bone marrow blast cells were reduced from >90% to near-normal levels. However, the contribution of a single dose of dexamethasone (40 mg IV given on Day 3) to the reduction in WBC count and blast cells is unknown. This patient's peripheral blood hematologic profile (WBC, absolute neutrophil, platelet counts and hemoglobin level) returned to near-normal, indicating a specificity of forodesine for leukemic cell populations and supporting that forodesine is an important breakthrough in the development of less toxic ALL therapy. Additional phase II results will be presented. [Display omitted]

How’s going on the T-Cell Brazil project

Lymphomas are a large, heterogeneous and infrequent group of neoplasms. Peripheral t-cell lymphomas (PTCL) is still rarer, and it makes up about 10-15% of lymphoid malignancies. Its prognosis is poor and remains a challenge. In April 2017, the first Brazilian collaborative effort was created to collect data from T-cell Lymphoma patients diagnosed in the five distinct macro-regions of Brazil. To better understand PTCL, according to the revised WHO-2017 classification, excluding Mycosis fungoides, Sézary syndrome, and chronic Lymphoproliferative disorders, all histological subtypes diagnosed from January 2015 to December 2022 will be enrolled. Tissue biopsied, immunophenotypic markers from consecutive patients at each site will be reviewed by panels of expert hematopathologists every year. So far, 381 cases were enrolled by 32 Brazilian centers, most of them are from the Southeast and South region. Twenty-two cases were not evaluable due to lack of complete data; hence, 359 were analyzed. The overall survival at 24-month was 52% (95% CI: 46-58%) and progression-free survival 37% (95% CI: 31-43%) with a median time of follow-up of 10 months (0.2-64). T-cell Brazil Project has many challenges to overcome, mainly due to its vast territory and its disparity. However, we already have another sixteen centers waiting for approval in their Ethics Committees. It was created a network around the country, mainly among the pathologists, with educational goals and exchange of experiences. It is crucial to obtain knowledge of local epidemiology to optimize resources, design clinical trials, and identify minority entities, contributing to increasing the quality o

Mycosis fungoides and S\xe9zary syndrome: a population-wide study on prevalence and health care use in Finland in 1998\u20132016

BackgroundInformation about health care use and costs of cutaneous T-cell lymphoma (CTCL) patients is limited, particularly in a European setting.MethodsIn this population-wide study we set out to investigate prevalence, and trends in health care use in two CTCL subtypes, mycosis fungoides (MF) and Sézary syndrome (SS) over a time period of 19 years in 1998–2016 by using a nation-wide patient register containing data on all diagnosed MF and SS cases in Finland.ResultsThe prevalence of diagnosed MF and SS rose from 2.04 to 5.38/100000, and from 0.16 to 0.36/100000 for MF and SS respectively during 1998–2016. We found a substantial decrease in inpatient treatment of MF/SS in the past two decades with a mean of 2 inpatient days/patient/year due to MF/SS in 2016, while the mean numbers of MF/SS related outpatient visits remained stable at 8 visits/year/patient. Most MF/SS-related outpatient visits occurred in the medical specialty of dermatology. In a ten-year follow-up after MF/SS diagnosis, the main causes for outpatient visits and inpatient stays were MF/SS itself, other cancers, and other skin conditions. Also cardiovascular disease and infections contributed to the number of inpatient days. Mean total hospital costs decreased from 11,600 eur/patient/year to 3600 eur/patient/year by year 4 of the follow-up, and remained at that level for the remainder of the 10-year follow-up. MF/SS accounted for approximately half of the hospital costs of these patients throughout the follow-up.ConclusionsThe nearly 3-fold increase in prevalence of diagnosed MF/SS during 1998–2016 puts pressure on the health care system, as this is a high-cost patient group with a heavy burden of comorbidities. The challenge can be in part answered by shifting the treatment of MF/SS to a more outpatient-based practice, and by adapting new pharmacotherapy, as has been done in Finland.

Venous thromboembolic events in T-cell lymphoma patients: Incidence, risk factors and clinical features

IntroductionPatients with hematologic malignancies are prone to developing thromboembolism. The incidence, risk factors and clinical features for developing venous thromboembolism (VTE) are not well-elucidated in patients with T-cell lymphoma. Materials and MethodsThe present study retrospectively analyzed 668 patients with VTE, including pulmonary embolism (PE) and deep vein thrombosis (DVT), who were admitted to Tianjin Medical University Cancer Institute and Hospital and Sun Yat-sen University Cancer Center from January 2006 to December 2018. All patients were diagnosed with T-cell lymphoma, and all episodes of symptomatic VTE were confirmed by imaging and ultrasound. The follow-up results were obtained through telephone communication and outpatient visits. ResultsA total of 668 patients were analyzed. Thirty-three (4.94%) patients had at least one episode of VTE, and all of which were deep vein thrombosis alone. All VTEs occurred in patients who received chemotherapy, while no VTE occurred in patients who did not receive chemotherapy. By univariate analysis, central venous catheter (CVC) (odds ratio [OR] 6.63, confidence interval [CI] 2.24−19.57, P = 0.001), Eastern Cooperative Oncology Group (ECOG) performance status 2, 3, or 4 (OR: 62.15, CI: 15.42−250.48, P = 0.000), and stage III or IV (OR: 4.06, CI: 1.00−16.40, P = 0.049) were identified as risk factors for developing VTE. By multivariate analysis, CVC (OR: 3.23, CI: 1.49−7.23, P = 0.003) and stage III or IV (OR: 2.30, CI: 1.06−4.97, P = 0.035) were still significant risk factors for developing VTE. ConclusionThe incidence of VTE in the present study population was comparable to that of lymphoma patients, other than T-cell lymphoma, and VTE was associated with CVC and advanced stage.

Correlation of mutational landscape and survival outcome of peripheral T-cell lymphomas

ObjectiveTo explore the correlation of mutation landscape with clinical outcomes in patients with peripheral T-cell lymphoma (PTCL).MethodsWe retrospectively analyzed the clinicopathological and prognosis data of 53 patients with PTCL from November 2011 to December 2017. Targeted next-generation sequencing of a 659-gene panel was performed for tissues from 53 patients with PTCLs. The correlation of mutation landscape with clinical outcomes was analyzed.ResultsTET2 was the most frequently mutated gene (64%), followed by RHOA (43%), PCLO (23%), DNMT3A (19%), IDH2 (17%), PIEZO1 (17%) and TP53 (15%). When mutated genes were categorized into functional groups, the most common mutations were those involved in epigenetic/chromatin modification (75%), T-cell activation (74%), and the DNA repair/TP53 pathway (64%). TET2/TP53 mutations were significantly associated with positive B symptoms (P = 0.045), and elevated lactate dehydrogenase (LDH) level (P = 0.011). Moreover, TET2/TP53 mutation was a risk factor for PTCL patient survival (HR 3.574, 95% CI 1.069 − 11.941, P = 0.039). The occurrence of JAK/STAT pathway mutations in angioimmunoblastic T-cell lymphoma (AITL) patients conferred a worse progression-free survival (HR 2.366, 95% CI 0.9130–6.129, P = 0.0334).ConclusionsHeterogeneous gene mutations occur in PTCL, some of which have a negative impact on the survival outcome.

Tamoxifen enhances romidepsin-induced apoptosis in T-cell malignant cells via activation of FOXO1 signaling pathway

Although romidepsin as monotherapy appears to be useful for treating T-cell lymphoma, combined chemotherapy with other therapeutic agents is required for improvement of the treatment outcome. To establish safer and more effective regimens, systematic screening was conducted to identify suitable drugs to be used in combination with romidepsin for T-cell malignancies, and the underlying molecular mechanisms were examined. The most effective agent was tamoxifen. The combination of romidepsin and tamoxifen had a significant synergistic effect in inducing apoptosis. The growth-inhibitory effects of the combined treatment were reversed by α-tocopherol. FOXO1 expression was greatly upregulated in MOLT-4 cells treated with romidepsin plus tamoxifen. Knockdown of FOXO1 expression by siRNA significantly reduced the cell death induced by romidepsin plus tamoxifen. The combination of romidepsin and tamoxifen might be considered for the treatment of T-cell lymphoma patients.

Programmed Cell Death Protein 1/Programmed Cell Death Ligand-1 Axis activates Intracellular ERK Signaling in Tumor Cells to Mediate Poor Prognosis in T-cell Lymphoma.

Purpose: To investigate the prognostic significance of programmed cell death ligand-1 (PD-L1) and phosphorylated ERK (p-ERK) and their interactions in T-cell lymphoma (TCL).Methods: The mRNA levels of PD-L1 and ERK in TCL samples were analyzed. Formalin-fixed paraffin-embedded tissues from 69 TCL patients were collected to detect the expression of PD-L1 and p-ERK by multiplexed immunofluorescence staining. The total PD-L1 and p-ERK was measured by western blotting, and membrane PD-L1 was determined using flow cytometry.Results: PD-L1 and ERK mRNA levels were significantly upregulated in TCL. The expression rates of PD-L1 and p-ERK were 52.2% and 27.5%, respectively. PD-L1 expression correlated with stage (R=0.304, P=0.011) and IPI score (R=0.313, P=0.009), and p-ERK expression correlated with stage (R=0.330, P=0.006) and IPI score (R=0.376, P=0.002). PD-L1 expression positively correlated with p-ERK expression (R=0.355, P=0.003). Patients with co-expression of PD-L1 and p-ERK had the worst overall survival (P=0.007). In three TCL cell lines with PD-L1 expression, we demonstrated that the expression of p-ERK was upregulated after stimulation with PD-1, suggesting that ERK signaling was activated.Conclusions: The PD-1/PD-L1 axis activates intracellular ERK signaling in tumor cells and that PD-L1, p-ERK or their combination are potential biomarkers for predicting the prognosis in TCL patients.