bluuebunny/crossref_metadata_2025_split huggingface dataset:
AFM13 Is the Most Advanced Bispecific NK-Cell Engaging Antibody in Clinical Development Substantially Enhancing NK-Cell Effector Function and Proliferation
Authors: Pahl Jens , Reusch Uwe , Gantke Thorsten , Kerber Anne , Koch Joachim , Treder Martin , Cerwenka Adelheid 
Published: October 2019

Abstract
Introduction:
AFM13 is an NK-cell engaging CD30/CD16A bispecific tetravalent TandAb antibody currently in phase 2 clinical development in Hodgkin lymphoma (HL) and other CD30+ malignancies. It engages NK-cells through CD16A with high affinity and specificity and confers significantly stronger NK-cell activation compared to other therapeutic antibodies. We have previously shown synergistic efficacy when NK-cell activation by AFM13 is combined with check-point modulation such as anti-PD-1 treatment, which is known to unleash T cell and NK-cell activity. The goal of this study was to identify further candidates for combination treatments and biomarkers that potentially indicate NK-cell responses to AFM13 treatment.
Methods:
AFM13-mediated NK-cell cytotoxicity and IFN-γ production after 4-hour interaction with HL cell lines was measured by 51Cr release assays and flow cytometry, respectively. Expression of NK-cell receptors, NK-cell proliferation (CFSE dilution) and expansion (absolute cell counts) was analyzed by flow cytometry.
Results:
The interaction of NK-cells with AFM13-coated tumor cells up-regulated the expression of NK-cell receptors such as CD25, CD69, CD137/4-1BB as well as molecules that may serve as NK-cell check-points when compared with the unrelated NK-cell binding TandAb AFM12 that does not bind to target cells. Importantly, CD16A engagement by AFM13 enhanced the proliferation and expansion potential of NK-cells when subsequently incubated with IL-15 or with particularly low doses of IL-2. NK-cell cytotoxicity and IFN-γ production was substantially increased towards CD30+ tumor cells in the presence of AFM13. Even target cells resistant to naïve and IL-2/IL-15-activated NK-cells were susceptible to AFM13-induced NK-cell cytotoxicity. AFM13 concentrations of as low as 10-2 µg/mL resulted in maximal activity while AFM13 was significantly more potent than native anti-CD30 IgG1 antibody. NK-cell activation by IL-2 or IL-15 had a synergistic effect on AFM13-mediated cytotoxicity.
Conclusion:
AFM13 specifically enhances the cytotoxic, proliferative and cytokine-producing potential of NK-cells. Our data indicate that the distinctive modulation of NK-cell receptors can be utilized to monitor NK-cell responses during AFM13 therapy and provides candidates for therapeutic combination strategies. Moreover, the combination with low doses of IL-2 or with IL-15 may expand the quantity of tumor-reactive NK-cells after AFM13 treatment and promote NK-cell functionality in the tumor microenvironment in cancer patients.

Disclosures
Reusch: Affimed: Employment, Patents & Royalties: Patents. Gantke:Affimed GmbH: Employment. Kerber:Affimed: Employment. Koch:Affimed: Employment. Treder:Affimed: Employment. Cerwenka:Affimed: Research Funding.
Arginase 1 Is a Marker of Myeloid-Mediated Immunosuppression with Prognostic Meaning in Classic Hodgkin Lymphoma
Authors: Romano Alessandra , Parrinello Nunziatina L , Tibullo Daniele , Vetro Calogero , Giallongo Cesarina , La Cava Piera , Chiarenza Annalisa , Villari Loredana , Tripodo Claudio , Motta Giovanna , Caruso Laura , Gallamini Andrea , Pileri Stefano A , Di Raimondo Francesco 
Published: October 2019

Abstract
Purpose : Neutrophilia is hallmark of classic Hodgkin Lymphoma (cHL), but its precise characterization remains elusive. We aimed at investigating the immunosuppressive role of high-density neutrophils in HL.
Experimental design : First, N-HL function was evaluated in vitro, showing increased arginase (Arg-1) expression and activity compared to healthy subjects. Second, we measured serum level of Arg-1 (s-Arg-1) by ELISA in two independent, training (N=40) and validation (N=78) sets.
Results : s-Arg-1 was higher in patients with advanced stage (p=0.045), B-symptoms (p=0.0048) and a positive FDG-PET scan after two cycles of chemotherapy (PET-2, p=0.012). Baseline levels of s-Arg-1 >200 ng/mL resulted in 92% sensitivity and 56% specificity to predict a positive PET-2.
Patients showing s-Arg-1 levels > 200 ng/mL had a shorter progression free survival (PFS). In multivariate analysis, PET-2 and s-Arg-1 at diagnosis were the only statistically significant prognostic variables related to PFS (respectively p=0.0004 and p=0.012).
Moving from PET-2 status and s-Arg-1 level we constructed a prognostic score to predict long-term treatment outcome: low s-Arg-1 and negative PET-2 scan (score 0, N=63), with a 3-Y PFS of 89.5%; either positive PET-2 or high s-Arg-1 (score 1, N=46) with 3-Y PFS of 67.6%, and both positive markers (score 2, N=9) with a 3-Y PFS of 37% (p=0.0004).
Conclusions : We conclude that N-HL are immunosuppressive through increased Arg-1 expression, a potential novel potential biomarker for HL prognosis.

Disclosures
Vetro: MLL Munich Leukemia Laboratory: Employment. Chiarenza:Gilead: Consultancy; Janssen: Consultancy; Roche: Consultancy.
Treatment of Primary Cutaneous Marginal Zone B-Cell Lymphomas with Intralesional Rituximab
Authors: Domingo Domenech Eva , Salas Queralt , Servitje Octavio , Climent Fina , Pomares Helena , Lucas Ana , Aguilera Carmen , Comai Alessandra , Mercadal Santiago , Gonzalez Barca Eva , Sureda Anna 
Published: October 2019

Abstract
BACKGROUND
Primary cutaneous marginal zone B-cell lymphomas (PCMZL) are low-grade lymphomas with an indolent course, but with a high rate of skin relapses. Surgical excision, local radiotherapy, or combinations of both are the first-line treatment in solitary or localized disease. Therapeutic options in cases of multifocal disease or relapse, include steroids or intravenous Rituximab in monotherapy or in combination with intravenous or oral chemotherapy. The effectiveness of intralesional Rituximab (ILR) in primary cutaneous B-cell lymphomas has been described in a small multicenter series of patients.
OBJECTIVE
To evaluate the effectiveness and safety of ILR in patients with PCMZL.
METHODS
In our center ILR is used since 2010 as fist-line treatment for patients with PCMZL with single facial lesions, multifocal disease that cannot be irradiated or in relapsed disease. The treatment regimen consist of a course of three injections of 10 mg (days 1, 3 and 5) in a single week at monthly intervals. Oral paracetamol and dexclorfeniramine were administrated 30 minutes before the procedure. Courses of ILR were monthly repeated until response or in cases of relapse. The primary outcome measure of the study was the rate of remissions (complete and partial remissions).
RESULTS
Twelve patients with PCMZL have been treated so far, 50% as first-line. The relapsed-group had previously received a median of 3 previous treatments (range, 1-6) including radiotherapy in 5 cases, surgery in 1 case, intravenous chemotherapy in 2 cases, and topic steroids in 2 cases. Clinical characteristics of the 12 patients are shown in the table below.
A total of 42 courses of ILR have been completed. All patients responded: a complete therapeutic response was achieved in 10 patients, whereas a partial response was observed in 2. No cases of progression or relapse in the site of ILR treatment were documented.
Four patients experienced transitory mild perilesional erythema that did not require medication and resolved in less than 4 hours. No cases of significant adverse events occurred and none of the patients suffered severe bacterial or opportunistic infections.
CONCLUSIONS
In our preliminary experience, ILR is an effective and safe therapeutic approach for selected PCMZL patients with single facial lesion, multifocal disease than cannot be irradiated, or in relapsed cases. To date, this is the largest experience with ILR of a single center.

Disclosures
Gonzalez Barca: Roche: Speakers Bureau; Janssen: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Gilead: Speakers Bureau.
Drug Shortage: The Impact of Substituting Mechlorethamine in Stanford V for Hodgkin's Lymphoma
Authors: Monteiro Luis P , Gaspar Catia L , Fevereiro Margarida , Tome Ana L , Almeida Nuno , Monteiro Alexandra , Sousa Aida B. 
Published: October 2019

Abstract
Introduction: the shortage of some anticancer drugs has forced their substitution in established combination chemotherapy regimens, but the replacement may have critical effects on the results of treatment and need careful evaluation. Since 1996 we treat Hodgkin's lymphoma (HL) with the Stanford V (SV) regimen, which achieves the same results as the more widely employed ABVD regimen with the benefits of a shorter duration (12 weeks) and lower cumulative doses of bleomycin and doxorubicin. When mechlorethamine became unavailable in 2008, we substituted cyclophosphamide at a dose of 650 mg/m2 for mechlorethamine 6 mg/m2. In 2012 the Pediatric Hodgkin Lymphoma Consortium, which had adopted the same policy, suggested in a retrospective analysis that the modification could lead to an inferior event-free survival (EFS). To assess the impact of the substitution, we retrospectively compared our results with the original SV (SVo) and the modified SV (SVm).
Methods: we evaluated the 265 patients consecutively treated in our center with SV between Nov/96 and Dec/14. We compared the pre-treatment characteristics, the complete remission (CR) rate, EFS and overall survival (OS) of the 184 patients who received SVo to those of the 81 patients treated with SVm. Radiotherapy indications and supportive measures were unchanged between the 2 periods.
Results: median age of the 2 groups was identical: 31 (16-69) for SVo and 31 (15-72) for SVm. No differences were found in histologic subtypes (nodular sclerosis 76/74%), frequency of B symptoms (36/47%, p=0.09), proportion of advanced stages (32/33%) or International Prognostic Score (≥3 in 20/13%, p=0.22). The CR rate was 89% for SVo and 86% for SVm. With a median follow up of 9 years for the SVo group and 4.6 years for the SVm group, there were no significant differences in EFS (76/73%, p=0.66) and OS (89%/85%, p=0.63) at 5 years.
Conclusion: the impact of forced substitutions in components of combination chemotherapy regimens due to drug shortage must be carefully scrutinized, notably in first line protocols for curable diseases. In the case of HL treated with the SV regimen, the present study is the first, to our knowledge, to evaluate the impact of replacing mechlorethamine by cyclophosphamide in adults. We found no difference in OS nor, unlike the pediatric study, in EFS. The substitution can be considered safe, since the modified version of SV achieves the same favorable results as the original SV.

Disclosures
No relevant conflicts of interest to declare.
NR4A3 Suppresses Lymphomagenesis In Vitro and In Vivo by Induction of Similar Patterns of Pro-Apoptotic Genes like NR4A1
Authors: Deutsch Alexander JA , Rinner Beate , Pichler Martin , Fechter Karoline , Greinix Hildegard T. , Wang Chenguang , Beham-Schmid Christine , Neumeister Peter 
Published: October 2019

Abstract
The nuclear orphan receptors NR4A1 and NR4A3 have been demonstrated as cooperating tumour suppressor genes leading to rapid development of acute myeloid leukaemia (AML) in double knock-out mice. In humans, their expression is reduced in leukemic blasts in AML patients. Furthermore, NR4A1 and NR4A3 hypoallelic mice develop preleukemic myelodysplastic/myeloproliferative disorders with progression to AML in some cases. Recently we published a comprehensive study of NR4A nuclear receptor expression levels in lymphoid neoplasms that revealed a marked reduction of NR4A1 and NR4A3 in the majority of patients with B-cell chronic lymphocytic leukaemia, with follicular lymphoma, and with diffuse large B cell lymphoma. Interestingly, functional characterization demonstrated that NR4A1 induces apoptosis of aggressive lymphoma cells in vitro and suppresses tumour growth in a xenograft mouse model.
Since the role of NR4A3 in aggressive lymphomas is unknown, we aimed to investigate its etiopathogenic function in these tumors. Low expression of NR4A3 was associated with poor survival in aggressive lymphoma patients. Experimentally, induction of NR4A3 expression by inducible ectopic expression in a variety of lymphoma cell lines led to a significantly higher proportion of apoptotic cells as demonstrated by DNA cleavage, Annexin V staining and increased caspase 3/7 activity. To test the tumor suppressor functions of NR4A3 in vivo, the stably transduced SuDHL4-lymphoma cell line was xenografted in the NOD-SCID-gamma (NSG) mouse model. In this system NR4A3 expression abrogated tumor growth in the NSG mice, whereas vector control and uninduced cells formed massive lymphoid tumors. Pharmacological activation of NR4A3 by Thapsigargin and BF175 resulted in a NR4A3 dependent induction of apoptosis in vitro. To dissect differential transcriptional activity of NR4A3 and NR4A1,both factors were separately over-expressed in four different aggressive lymphoma cell lines followed by semi-quantitative mRNA expression analysis of intrinsic and extrinsic apoptotic genes. NR4A1 or NR4A3 over-expression caused apoptosis by induction of BAK, Puma, BIK, BIM, BID and Trail to the same degree.
In summary, our data suggest that NR4A3 possesses tumor suppressive function in aggressive lymphomas by pro-apoptotic transactivation and that NR4A3 is functional redundant to NR4A1 in aggressive lymphomas.

Disclosures
No relevant conflicts of interest to declare.
Dnmt3a Haploisufficiency Cooperates with Oncogenic Kras to Promote an Early-Onset T-Cell Leukemia
Authors: Chang Yuan-I , Kong Guangyao , Zhang Jing , Ranheim Erik A. 
Published: October 2019

Abstract
Recent whole genome/exome sequencing efforts in myeloid malignancies identified that mutations in DNA methyltransferase 3A (DNMT3A) are prevalent in acute myeloid leukemia (AML). In addition, DNMT3A mutations are also identified in various T cell malignancies. Of note, DNMT3A mutations are typically heterozygous and some WT DNMT3A functions thus remain in this state. However, the predominant DNMT3A R882 mutations, which locate in the catalytic domain, seem to inhibit the methyltransferase activity of the remaining WT DNMT3A due to its dominant-negative function (Yang L, Rau R, Goodell MA, Nat. Rev. Cancer 15: 152-165, 2015). COSMIC database analysis reveals different prevalence of DNMT3A R882 mutations in various hematopoietic malignancies. Approximately 60% of DNMT3A mutations in AML are R882 mutations, while the frequency of R882 mutations drops to ~40% in myelodysplastic syndrome (MDS) and myeloproliferative neoplasm (MPN). In contrast, the frequency of R882mutations is less than 25% in T-cell acute lymphoblastic leukemia (T-ALL). The significantly different frequencies of DNMT3A R882 mutations in AML versus T-ALL inspired us to investigate whether downregulation of DNMT3A regulates malignancies of different lineages in a dose-dependent manner.
We previously showed that Dnmt3a-/- promotes MPN progression in KrasG12D/+ mice and ~1/3 compound mice develop AML-like disease (Chang et al. Leukemia 29: 1847-1856, 2015). Here, we generated KrasG12D/+; Dnmt3afl/+; Mx1-Cre mice to determine how Dnmt3a haploisufficiency affects KrasG12D/+-induced leukemogenesis. After pI-pC injections to induce Mx1-Cre expression, primary KrasG12D/+; Dnmt3a+/- mice died quickly as primary KrasG12D/+ mice; the survival rates of these two groups of animals were not significantly different. However, in a competitive transplant setting, recipients transplanted with KrasG12D/+; Dnmt3a+/- bone marrow cells displayed a significantly shortened survival than recipients with KrasG12D/+ cells. Moreover, all of the recipients with KrasG12D/+; Dnmt3a+/- cells developed a lethal T-ALL without significant MPN phenotypes, while ~20% of recipients with KrasG12D/+ cells developed MPN with or without T-ALL. This is in sharp contrast to the recipients with KrasG12D/+; Dnmt3a-/- cells, in which ~60% developed a lethal myeloid malignancy (MPN or AML). Our data suggest that in the context of oncogenic Kras, loss of Dnmt3a promotes myeloid malignancies, while Dnmt3a haploisufficiency induces T-ALL. This dose-dependent phenotype is highly consistent with the prevalence of DNMT3A R882 mutations in AML versus T-ALL in human. We are currently investigating the underlying mechanisms.

Disclosures
No relevant conflicts of interest to declare.
GDPT Versus CHOP in Newly Diagnosed Peripheral T-Cell Lymphoma: A Prospective Randomized Controlled, Open-Label Study (No.NCT01664975)
Authors: Li Ling , Duan Wenjing , Young Ken H. , Li Zhaoming , Zhang Lei , Fu Xiaorui , Li Xin , Sun Zhenchang , Zhang Xudong , Yan Jiaqin , Nan Feifei , Chang Yu , Lin Li , Zhang Mingzhi 
Published: October 2019

Abstract
Background: Peripheral T-cell lymphoma is a distinct lymphoid neoplasm with aggressive course and poor outcome. Optimal treatment strategies for peripheral T-cell lymphoma have not been well defined. We compared the efficacy and safety of GDPT and CHOP regimens for patients with newly diagnosed peripheral T-cell lymphoma in a prospective randomized controlled and open-label clinical trial (No.NCT01664975).
Methods: All eligible patients with newly diagnosed peripheral T-cell lymphoma had measurable disease with an ECOG performance status ≤ 2 and adequate organ function. GDPT or CHOP chemotherapy were randomly assigned to patients. Patients in arm GDPT received intravenous gemcitabine (0.8g/m2) in 30 min on days 1 and 8, cisplatin (25mg/m2) on days 1-3, and oral prednisone (60 mg/m2) on days 1-5, thalidomide (200 mg) until the end of the whole chemotherapy. Patients in group CHOP received intravenous cyclophosphamide (750 mg/m2), doxorubicin(50 mg/m2) and vincristine (1.4 mg/m2, maximum 2 mg) on day 1, and oral prednisone (60 mg/m2) on days 1-5. Each cycle was repeated six times every 3 weeks. Efficacy was evaluated every two cycles. The primary endpoint was to evaluate the efficacy assessed by progression-free survival. Secondary end points included response rate and overall survival.
Results: Between July 2010 and June 2016, 103 patients allocated into two groups randomly, of whom 52 were treated with GDPT therapy and 51 were treated with CHOP therapy. Patient characteristics were well balanced within the two arms of treatment at enrollment (Table 1). The 2-year progression-free survival (PFS) and overall survival (OS) rates were better in GDPT group than that in CHOP group (57% versus 35% for 2-year PFS, P=0.0035; 71% versus 50% for 2-year OS, P =0.0001) (Figure 1). Complete remission (CR) rate and overall response rate (ORR) of GDPT group were higher than that in CHOP group (52% versus 33%, P =0.044 for CR rate; 67% versus 49%, P =0.046 for ORR) (Table 2). Adverse effects of chemotherapy was hemocytopenia predominantly in both arms. No differences were observed between the two arms in terms of grade 3/4 myelosuppression, digestive tract, hepatic, renal, cardiac or neurological toxicity (Table 3). Acute toxicity was moderate, tolerable and well managed in both arms.
Conclusions: GDPT chemotherapy resulted in significant improvement in PFS and OS compared with CHOP chemotherapy and side effects of chemotherapy was well tolerated for newly diagnosed peripheral T-cell lymphoma patients. Therefore, GDPT is a promising new regimen as potential first-line therapy against peripheral T-cell lymphoma.

Disclosures
No relevant conflicts of interest to declare.
Long-Term Follow-up of Primary Bone Diffuse Large B-Cell Lymphoma Treated with m NHL-BFM-90
Authors: Gabeeva Nelly G. , Zvonkov Eugene E. , Morozova Anna K. , Koroleva Daria A. , Kovrigina Alla M. , Savchenko Valery Grigorievich 
Published: October 2019

Abstract
Background: Primary bone lymphoma (PBL) accounts for about 5% of extranodal lymphomas, the most common type are Diffuse Large B-cell Lymphoma (PBL-DLBCL). R-CHOP chemotherapy combined with or no radiation therapy can induce 5-year overall and progression free survival about 63% and 60% for localized-stage and only 41% and 35% for advanced stage and with adverse factors (AF). Due to the rarity of the disease the optimal treatment strategy still remains unknown. Further research is thus needed to increase treatment efficacy for patients with advanced stage PBL-DLBCL and with AF.
Aims: To evaluate the efficacy of mNHL-BFM-90 program in adult patients with advanced stage PBL-DLBCL and with AF.
Methods: Twenty three previously untreated pts with PBL-DLBCL underwent mNHL-BFM-90 treatment between May 2007 and July 2014; mean age 41 years (range 17-65); age ≥60 years 3 pts (13,6%); M\F=14\9. Tumor stage according to the Ann-Arbor classification: stage I (involvement of 1 bone) - 7 pts (30,4%), II (involvement of regional lymph nodes) - 3 pts (13%), IV (multiple bone involvement) - 13 pts (56,6%); stage >I 16 pts (69,5%). All pts with stage I had large-size tumors (>6sm) with soft tissue contact invasion. LDH elevation was observed in 10 pts (43,5%), B-symptoms in 15 pts (65%), large-size tumors - 20 pts (87%). The most common tumor sites were the skull bones, vertebrae, and femur. All patients received treatment according to the mNHL-BFM-90 intensive program This program was modified in the following way: adriamycin was administered on the 3rd day of course AA at a dose 50 mg/m2; methotrexate was administered on the 1st day of course C at a dose 1.0 g/m2 for 12 hours. All patients underwent from 4 to 6 courses. Twelve (52%) pts received rituximab on day 0 prior to each course of therapy. Five pts received radiotherapy, all pts received intrathecal prophylaxis of central nervous system (CNS) involvement.
Results. Complete remissions were achieved in 23 pts (100%). Relapses were observed in three patients in 6, 9 and 40 months after completion of treatment. From 2 pts with early relapse, one was diagnosed as «double-expression lymphoma» (MYC expression >70%, BCL2 >60%), both pts proliferation index Ki-67 accounted 100%. All 3 pts received salvage therapy and died from progression of disease.
With a median follow-up of 69 months (range 17-110) both progression-free and overall survival of 23 pts constituted 87%. Grades 3, 4 hematological toxicity was observed in all pts. No deaths related to mNHL-BFM-90 treatment occurred. No second malignancies were observed.
Conclusion The mNHL-BFM-90 demonstrated high efficacy and acceptable toxicity in patients with advanced-stage PBL-DLBCL and with AF.
Figure Figure.

Disclosures
No relevant conflicts of interest to declare.