bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of Mitochondrial Apoptotic Pathway in Disulfiram/Copper Mixture-Induced Cell Apoptosis in Human B-Lineage Acute Lymphoblastic Leukemia in Vitro
Authors: Xu Bing , Deng Manman , Jiang Zhiwu , Li Jie , Chen Kai , Li Peng
Published: November 2019

Backgrounds: The long term prognosis of adult B-lineage acute lymphoblastic leukemia (B-ALL) is poor when compared with pediatric B-ALL. The current therapeutic regimen for adult B-ALL often results in refractory and relapsing diseases. Therefore, it is urgently needed to explore novel approaches to treat adult B-ALL. Disulfiram (DS) has been used clinically as a safe anti-alcoholism drug for over 6 decades. Recent studies demonstrated that disulfiram/cooper mixture (DS/Cu) was cytotoxic to multiple solid cancers, but its effects on B-ALL cells are still unclear. Moreover, the molecular mechanism of the cytotoxicity of DS/Cu to tumor cells was poorly defined. In this study, we investigated the effects of DS/Cu on B-ALL cells in vitro and its related cytotoxic mechanism.

Results: Firstly, CCK8 assay indicated that DS/Cu markedly inhibited Nalm6 cell proliferation in a dose-dependent manner. Secondly, colony-forming assay showed that DS/Cu also abolished the clonogenicity of Nalm6 cells (P<0.001). Thirdly, FACS analyses revealed that DS/Cu mixture could induce apoptosis of Nalm6 cells, as well as primary B-ALL cells (n=16) in a dose-dependent manner. We additionally analyzed the relationship between clinical characteristics of B-ALL patients, including age, WBC counts, immunophenotype, cytogenetics, risk stratification and Ph chromosome, with the efficacy of DS/Cu on B-ALL cells. The apoptosis isolated from pro-B and cytogenetic abnormality B-ALL pastients was higher. Therefore, our results demonstrated that DS/Cu mixture could induce significant cytotoxicity against B-ALL cells in vitro.

To decipher the cytotoxic mechanism of DS/Cu mixture, JC-1 staining was done and the results showed that DS/Cu mixture could significantly reduce the mitochondrial membrane potential in Nalm6 cells (P<0.01) and 7 cases of primary B-ALL cells (P<0.05). Consistently, Western Blot analysis showed that DS/Cu induced B-ALL cell apoptosis by down-regulating the expression of anti-apoptotic protein Bcl2 and Bcl-XL, as well as activating caspase-3 and its substrate PARP. Hence, our results indicated that DS/Cu induced apoptosis of B-ALL cells at least partly through the intrinsic mitochondrial apoptotic pathway.

Conclusion: Our results demonstrated that DS/Cu not only significantly inhibit proliferation and clonogenicity, but also induce apoptosis of B-ALL cells in vitro.The mitochondrial apoptotic pathway might be the molecular mechanism of DS/Cu-induced apoptosis of B-ALL cells.

Disclosures

No relevant conflicts of interest to declare.
Drug-DNA Conjugated Gold Nanoparticles for the Treatment of Acute Myeloid Leukemia
Authors: Gossai Nathan , Naumann Jordan , Zamora Ed , Li Nan-Sheng , Piccirilli Joseph , Gordon Peter M.
Published: November 2019

INTRODUCTION: Novel therapies are needed for acute myeloid leukemia as only ~60% of children are cured despite maximally intensive cytotoxic chemotherapy. Functionalized gold nanoparticles (AuNP) are utilized for many biomedical applications and represent a potentially novel therapeutic approach in leukemia. We enhanced this technology by developing an AuNP system that selectively releases molecularly targeted drugs in leukemia cells.

METHODS: AuNPs were functionalized with short, double-stranded oligonucleotides with sequence complementarity to genes overexpressed in or unique to a leukemia cell (e.g. survivin or AML/ETO). Only the anti-sense oligonucleotide is covalently bound to the nanoparticle via a thiol linker. Thus, after entering leukemia cells, the endogenous targeted oncogene mRNA can bind to its complementary sequence on the AuNP and displace the non-covalently bound oligonucleotide which, in our system, is conjugated to the multi-tyrosine kinase inhibitor dasatinib. As a binary reaction, the amount of dasatinib-conjugated oligonucleotide released from the nanoparticle is directly proportional to both the presence and abundance of the complementary mRNA present in a cell. We evaluated AuNP uptake into multiple AML cell lines, as well as normal hematopoietic cells. The effect of dasatinib-AuNPs on dasatinib-sensitive leukemia cell lines was evaluated using proliferation assays, annexin V staining, and cell colony assays. Toxicity in T-cells and CD34+ cells was assessed with T-cell activation and p-SRC assays, respectively.

RESULTS: Conjugation of dasatinib to an oligonucleotide complementary to a region of the survivin gene did not perturb its ability to inhibit SRC and c-KIT kinases in vitro. Leukemia cells demonstrate highly efficient AuNP uptake when cultured alone or with up to a 100-1000 fold excess of normal bone marrow cells. Treatment of K562 leukemia cells, containing a BCR/ABL translocation and high levels of survivin mRNA, with dasatinib-AuNPs resulted in dose-dependent p-SRC and p-CRKL inhibition. Furthermore, K562 cells also showed significantly impaired proliferation, increased apoptosis, and formed fewer colonies in methylcellulose. Conversely, normal T-cells and CD34+ cells, which express less survivin than leukemia cells, were significantly less affected by dasatinib-AuNPs than dasatinib alone as measured by T-cell activation assays and p-SRC levels, respectively.

CONCLUSIONS: This method of using functionalized AuNPs to deliver and activate dasatinib in leukemia cells augments drug efficacy while minimizing toxicity and thus represents a novel therapeutic strategy for AML. Ongoing work is underway to characterize the mechanism of preferential AuNP uptake in leukemia cells and assessing in vivo activity using a murine xenotransplantation model of leukemia.

Disclosures

No relevant conflicts of interest to declare.
Thioridazine Enhances TRAIL Cytotoxicity in Human Myeloid Leukemic Cells By up-Regulating DR5 and Modulating PI3K-AKT-NF-Kb Pathway
Authors: Wang Yi , Bao Yangyi , Xia Leiming , Liu Liu , Guo Kunyuan , Yang Mingzhen
Published: November 2019

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can induce apoptosis in cancer cells but not in most normal cells, and is identified to be effective in various cancers, include myeloid leukemic cells[1]. Although some leukemia cell lines, K562 and KG-1, are sensitive to TRAIL, many showed certain degrees of resistance to TRAIL-mediated apoptosis[2,3], and the mechanism remains largely unknown, which forced us to find out ways to solve the problem.

In this study, we investigated whether thioridazine, a phenothiazine derivative, could overcome the TRAIL resistance in K562 and KG-1 cells. Recently, we showed that Compared to treatment with thioridazine or TRAIL alone, co-treatment with thioridazine and TRAIL-induced apoptosis in K562 and KG-1 cells synergistically. This combination led to activation of caspase-8 and Bid, the cytosolic cumulation of cytochrome c from mitochondria as well as caspase-3 activated downstream. Treatment with thioridazine induced down-regulation of PI3K-AKT-NF-κB pathway. meanwhile, thioridazine dropped the level of NF-κB-dependent Bcl-xL, leading caspase activated and Bid cleaved. the expression of TRAIL-receptors in both K562 and KG-1 cells underwentthe treatment of thioridazine investigated that thioridazine significantly up-regulated DR5 by up to 51.22%, but not other TRAIL-receptors such as DR4, decoy receptor 1, and DcR2.

Therefore, our results indicate that the combination of TRAIL with thioridazine overturn TRAIL resistance through Up-regulating the expression of DR5 and down-regulation of AKT protein, and combination treatment with thioridazine and TRAIL may be a novel therapeutic strategy in leukemia.

Reference: Srivastava R K. TRAIL/Apo-2L: mechanisms and clinical applications in cancer.[J]. Neoplasia, 2001, 3(6):535-546. Nimmanapalli R, Porosnicu M, Nguyen D, et al. Cotreatment with STI-571 enhances tumor necrosis factor alpha-related apoptosis-inducing ligand (TRAIL or apo-2L)-induced apoptosis of Bcr-Abl-positive human acute leukemia cells.[J]. Clinical Cancer Research An Official Journal of the American Association for Cancer Research, 2001, 7(2):350-357. Yang T, Lan J, Huang Q, et al. Embelin sensitizes acute myeloid leukemia cells to TRAIL through XIAP inhibition and NF-κB inactivation.[J]. Cell Biochemistry & Biophysics, 2015, 71(1):291-297.

Disclosures

No relevant conflicts of interest to declare.
A Gene Panel NGS-Based Strategy for Genomic Characterization of Acute Myeloid Leukemias (AMLs)
Authors: Bernardi Simona , Cattina Federica , Di Palma Andrea , Borlenghi Erika , Schieppati Francesca , Perucca Simone , Cancelli Valeria , Turra Alessandro , Malagola Michele , Skert Crisitina , Filì Carla , Cattaneo Chiara , Passi Angela , Farina Mirko , Rossi Giuseppe , Mignone Flavio , Russo Domenico
Published: November 2019

AMLs are clonal disorders characterized by high genomic heterogeneity and several chromosomal and molecular alterations affecting patients' outcome. In about 40% of AML patients who do not show any citogenetic alteration, sequencing analysis identified different gene mutations which play a pivotal role in leukemogenesis and have a negative prognostic impact: FLT3, ASXL1, TET2, IDH1, IDH2, RUNX1, CBL, CEBPα, DNMT3A and TP53. Conventional Sanger sequencing may detect clones representing more than 20% of the total tumor population, whereas Next Generation Sequencing (NGS) can identify mutations in less than 1% of leukemic cell burden. The detection of these variants is relevant because they can play an important role in driving drug resistance and disease relapse and for biologic risk assessment.

To that purpose, we designed a 23-genes panel including: FLT3, DNMT3A, RUNX1, ASXL1, IDH1, IDH2, BCOR, NRAS, KRAS, TET2, TP53, U2AF1, ZRSR2, SF3B1, SRSF2, CBL, CEBPα, EZH2, NPM1, TERT, TERC, ETV6, GATA2. By using a 454 GS Junior by Roche Diagnostics with an amplicon-based sequencing approach and performing three sequencing runs per sample in order to reach a sensitivity of at least 1%, we settled an investigative strategy in order to assess the genomic profile of AMLs at diagnosis and possibly at the time of relapse or resistance. By this approach, we were able to confirm all the variants (i.e. FLT3, NPM1) previously documented with conventional tests; to reveal the presence of variants under the Sanger threshold of 20% in the genes resulted as wild type with routinely analysis; to evaluate the AML clonal heterogeneity, by assessing the coexistence of several mutated clones which have been described to be related to drug resistance, poor prognosis or to prior myelodysplastic syndrome.

This gene panel NGS-based strategy may be proposed as a highly accurate and sensitive approach for genomic characterization of acute myeloid leukemias and as an useful tool for planning a target and personalized AML therapy.

Disclosures

No relevant conflicts of interest to declare.
Post Induction WT1 MRD Analysis Significantly Predicts Early Relapse in Acute Myeloid Leukemia (AML) Patients
Authors: Capelli Debora , Attolico Imma , Mancini Giorgia , Mancini Stefania , Saraceni Francesco , Viola Nadia , Scortechini Anna Rita , Pascale Pasquina , Scortechini Ilaria , Leoni Pietro , Olivieri Attilio
Published: November 2019

AML patients early relapsing within 12 months, still represents an extremely poor prognosis setting. Cytogenetic and molecular data are not diagnostic in 20-25% of AML patients and intermediate 2 ELN risk category has still an undefined prognosis. Post induction and consolidation MRD might represent a new prognostic factor besides patients and disease characteristics. We have evaluated post induction and consolidation bone marrow mimimal residual disease (MRD) in 126 AML patients (median age: 61.5 years, range: 17-89) with 20 months median follow-up (range 2-79.9). We analysed abnormal leukemia immunophenotype (ALIP) by multiparameter flow cytometry (MPFC) and WT1 by RT-PCR as described by Buccisano et al and Cilloni et al. Cytogenetic, NPM and FLT3 status were performed in 111, 97 and 119 patients respectively, defining the molecular cytogenetic risk in 96 patients. WT1 was +ve in 91/114 patients (70%) at diagnosis (median 1,231.5; range: 2-268,784), in 11/71 (15.5%) post induction (median 1216; range:260-134,633) and in 8/66 (12,1%) post consolidation (median 627.8; range:258-45,338). MPFC MRD was +ve in 33/66 (50%) patients after induction and in 18/48 (37.5%) after consolidation. We analysed 12 month Cumulative Incidence of Relapse (CIR) adjusted by MRD status, patients and disease characteristics. 82/99 patients achieved CR, 40 relapsed in a median of 8 months (1-52 months) 29 within 12 months with 37.5% 1 yr CIR. Patients receiving chemotherapy (38), Autologous (14) and Allogeneic Transplant (39) as post consolidation treatment had 45%, 35% and 30% 1 yr CIR respectively. NPM+FLT3- patients had 25% 1 yr CIR, compared to 25% in NPM-FLT3-, 50% in NPM+FLT3+ and 47% in NPM-FLT3+. Patients with WT1 positive post induction and consolidation had 1 year CIR of 90% and 72.5% respectively. Patients with MPFC positive post induction and consolidation had a 1 year CIR of 48.6% and 44.5% respectively. Multivariate analysis identified post induction WT1 positive status as the main predictor of 1 year CIR. Patients with WT1 positive after induction had a a 15.8 RR of 1 year CIR(p<0.0001) in comparison to WT1 negative patients. Hyperleukocytosis (WBC count >50,000/ml at diagnosis) and age>60 yrs also significantly predicted 1 year CIR with a RR of 7.22 (p=0.002) and 9.1 (p=0.001) respectively. In conclusion WT1 post induction status confirmed its prognostic significance in our series targeting a subset of early relapsing patients with an extremely poor outcome deserving experimental approaches.

Disclosures

No relevant conflicts of interest to declare.
Pim-1, Beyond an Oncogene, in Acute Myeloid Leukemia
Authors: Wu Yu , Sun Ruixue , Lun Yan
Published: November 2019

Background and Objective:

There are 3 members in Pim family, Pim-1, Pim-2 and Pim-3. They are serine/threonine kinase coding proto-oncogene. It is reported that Pim-1 plays a role in solid tumor, leukemia and polycythemia vera, et al. Pim-3, as a newly cloned oncogene has discovered to functioning in hepatic carcinoma, pancreatic cancer, et al. We are trying to find out the roles of Pim1 in acute myeloid leukemia.

Methods:

1 Evaluate the expressions of Pim family genes in acute myeloid leukemia patients and analysis the profile of Pim expressions with clinical characteristics.

2 Up-regulating the expression of Pim1 in AML cell lines by transient transfection or long-term infection through GFP-expressing plasmids and lenti-virus system and analyze the proliferation, apoptosis and chemotaxis features of the transfected AML cell lines.

Results:

1 Our investigation showed that Pim-1 is up-regulated in around 15% of acute myeloid leukemia (AML) patients.

2 As shown in growth curves and apoptosis assays, over expression of Pim-1induces growth up-regulation and anti-apoptosis. We hypothesized that phenomenon could be originated from the enhanced expression of c-myc, cyclin D1 and Bad phosphorylation shown in western blotting analysis.

3 Our chemotaxis assay and bone marrow stromal cell co-culture model demonstrated that overexpression of Pim-1can enhance the AML cells chemotactic movement toward SDF-1¦Á, with calcium influx increment and phosphorylation of CXCR4.

4 Flow cytometry analysis and confocal immunofluorescence observation demonstrated the up-regulated CXCR4 expression and internalization induced by SDF-1¦Á.

5 We also checked the angiogenesis and adhesion molecule during the process but did not show any contribution.

Conclusion:

Pim-1, beyond as an oncogene, can promote growth or anti-apoptosis of AML cells, can interact with microenviroment through SDF-1¦Á-CXCR4 axis. The interplay between AML cells and microenviroment mediated by Pim-1 can make sense in AML target therapy and make a good adjunctive for transplantation conditioning regimen.

Figure 1. Expression of Pim1 in AML patients and control Figure 1. Expression of Pim1 in AML patients and control

Disclosures

No relevant conflicts of interest to declare.
Minimal Residual Disease Evaluation By Real-Time Quantitative Polymerase Chain Reaction for Wilms Tumor 1 Gene Expression and Multicolour Flow Cytometry, in Pediatric B Lineage Acute Lymphoblastic Leukemia
Authors: Varma Neelam , R. Sandeep , Naseem Shano , Sachdeva Man Updesh Singh , Bansal Deepak , Varma Subhash , Malhotra Pankaj
Published: November 2019

Introduction: Minimal residual disease (MRD) determination involves the measurement of very low levels of leukemia using sensitive techniques which at present are complex, time consuming and require expertise for performance and interpretation. A ''panleukemic''marker such as Wilm's tumor 1 gene (WT1) which is frequently over expressed in acute leukemia could simplify MRD detection and serve as a useful prognostic marker.

Aim of study: To evaluate the usefulness of WT1 gene expression, as a marker for MRD in B-lineage acute lymphoblastic leukemia (B-ALL).

Method and Material: Flow cytometric immunophenotyping (FCMI) and real time-polymerase chain reaction (RQ-PCR) for WT1 gene expression were performed usingbone marrow at diagnosis and at day 15 (mid-induction). Of the 23 patients recruited, day 15 MRD analyses by both these methods was performed on 11 bone marrow samples of patients who showed WT1 over expression at day 0.

Results: WT1 over expression at diagnosis was found in 69.5% cases (16/23). MRD was detectable in 54.5% cases by WT1 RQ-PCR and by FCM in 72% cases. A statistically significant correlation was seen between WT1 normalized copy number (NCN) at diagnosis with MRD levels detected by FCM.

Conclusion: WT1 represents a candidate MRD and prognostic marker. The significant correlation between WT1 over expression at diagnosis and MRD positivity by flow cytometry at day 15 (mid induction) of chemotherapy suggests that high WT1 expression could correlate with unfavourable outcome in childhood ALL.

However, as not all patients of B-ALL over-express WT1 at diagnosis, quantitative assessment of WT1 transcripts can be used as useful molecular marker for MRD detection, but only in a subset of patients.

Disclosures

No relevant conflicts of interest to declare.
Low Epression of MiR-18a Distinguishes Pediatric and Adult Acute Lymphoblastic Leukemia from Each Other
Authors: Mosakhani Neda , El Missiry Mohamed , Vakkila Emmi , Heikkilä Päivi , Knuutila Sakari , Vakkila Jukka
Published: November 2019

In several adult solid cancers the presence or absence of an inflammatory microenvironment has turned out to be an important prognostic factor. Acute lymphoblastic leukemia (ALL) is seen in both adults and children but the response to chemotherapy and survival is significantly worse in adults than children. Therefore, we wanted to study whether the expression of immune system associated molecular markers would be different in adult and pediatric ALL patients at the time of diagnosis. IDO and FOXP3 were studied from paraffin embedded tissue samples by immunohistochemistry in 12 pediatric and 10 adult bone marrow samples. Inflammation associated miRNA analysis were performed in 19 adult and 79 pediatric ALL patients and involved miR-10, miR-15, miR-16, miR-17-92 cluster, miR-33, miR-146a, miR-150, miR-155, miR-181a, miR-222, miR-223, and miR-339. miRNAs were first analysed by Agilent's miRNA microarray and thereafter validated by qRT-PCR. miRNAs not expressed in at least 75% of one group of samples were excluded. Significance (p <0.05; q<0.1) of differential expression was estimated by t-test for those miRNAs with at least a 2.0 fold change. Sufficient RNA for qRT-PCR was available for 42 pediatric and 19 adult patients. The adult and pediatric ALL patients had quantitatively and qualitatively similar expression of IDO and FOXP3 in leukemic bone marrow samples (p=0.26 and 0.74, respectively). Out of studied miRNAs only miR-18a differed significantly in microarray analysis between adult and pediatric ALL being lower in children (FC -3.74; p 0,0037). Results were confirmed by qRT-PCR (upregulated in adults, FC 3.71, p 0.003161). The other members of the miR-17-92 cluster did not differ significantly. We conclude that pediatric and adult ALL patients have remarkably similar pattern of immune cell associated markers in bone marrow at diagnosis. This is in line with recent evidence that the outcome of the adult ALL patients can be significantly improved if treated with pediatric protocols. However, the low expression of miR-18a in pediatric ALL is interesting and demands further studies.

Disclosures

No relevant conflicts of interest to declare.
Prognostic Value of Pleural Effusion in Hodgkin's Lymphoma Patients
Authors: Bhatt Nidhi , Brandt Gregory , Niebrugge Daniel , Khan Aziz U.
Published: November 2019

Background: There has been little investigation into the incidence or prognostic significance of a pleural effusion when present with the diagnosis of Hodgkin's Lymphoma. In our small clinical practice we had two patients with Hodgkin's lymphoma who presented with pleural effusion and subsequent poor outcomes, including relapse and eventual death. These events stemmed this retrospective study investigating the incidence of pleural effusion with the diagnosis of Hodgkin's disease and whether the presence of pleural effusion at the time of diagnosis would reveal any association with outcomes in a larger cohort of patients.

Procedure: A single centered retrospective chart review of 134 patients' age 0-50 with confirmed Hodgkin's lymphoma was performed. Charts were evaluated for the presence of any radiographic evidence of pleural effusion at the time of diagnosis, along with the patient's eventual outcome.

Result: 8/133 (6%) patients with Hodgkin's lymphoma had a pleural effusion at the time of diagnosis. Death is dependent on pleural effusion (p= 0.019) with 3 of 8 (37.5%) of patients that had pleural effusion who died.There is an 8.77 (CI 1.8 - 43.5) times greater odds of death for patients with pleural effusion versus those without pleural effusion.

Recurrence is dependent on pleural effusion (p= 0.0061) with 5/8 (62.5%) of patients that had pleural effusion who experienced recurrence.There is an 8.75 (CI 1.9 - 39.6) times greater odds of recurrence for patients with pleural effusion versus those without pleural effusion.

Of the 5 patients who had recurrence, as stated above 3 patients died, 1 patient was salvaged by stem cell transplant and another patient was recently diagnosed with relapse thus, clear outcome is yet to be determined

Conclusion: The presence of a pleural effusion at the time of diagnosis of Hodgkin's lymphoma is a rare finding, yet when present, was associated with death and recurrence. This finding may suggest a more aggressive initial treatment be required given the association with poor outcomes. An evaluation of a larger cohort of patients, along with a determination of the diagnostic stage, will be required to confirm prognostic potential.

Disclosures

No relevant conflicts of interest to declare.
Treating Hodgkin Lymphoma without Dacarbazine: A Lesson Learned from Thailand
Authors: Suwanban Tawatchai , Nakhakes Chajchawan , Lekhakula Arnuparp , Julamanee Jakrawadee , Norasetthada Lalita , Nawarawong Weerasak , Chansung Kanchana , Sirijerachai Chittima , Khuhapinant Archrob , Siritanaratanakul Noppadol , Bunworasate Udomsak , Intragumtornchai Tanin , Numbenjapon Tontanai , Prayongratana Kannadit , Nipharuck Pimjai , Chuncharunee Suporn , Wongkhantee Somchai , Wong Peerapon , Kanitsap Nonglak , Pornuipavee Rachanid , Makruasi Nisa
Published: November 2019

INTRODUCTION: Hodgkin lymphoma (HL) is curable if the patients are treated early and properly. Thailand is a country where public health care resources are limited and as such, dacarbazine was not listed in the national drug list, thus, cannot be reimbursed in patients under the Universal Coverage payment scheme which comprised 60-70% of all patients. This study was aimed to compare outcome differences between patients with HL who were treated with ABVD (doxorubicin, bleomycin, vinblastine, dacarbazine), ABV (doxorubicin, bleomycin, vinblastine) and COP/ABV (cyclophosphamide, vincristine, prednisolone, procarbazine /ABV) as happened in the real world practice in Thailand.

METHODS: From the nationwide multicenter registry of 4,371 newly diagnosed lymphoma patients in Thailand from 2007 to 2014, there were 316 patients with HL. The data is part of the Thai Lymphoma Study Group Registry performed at the thirteen major medical centers in Thailand. The patients' characteristics, treatment options and outcomes were analyzed.

RESULTS: The median age was 42 years (range,13-89 years). Male: female was 1.3:1. Fifteen percent of the patients were older than 60 years. Histological subtypes were classical HL, nodular sclerosis, 45.3%, mixed cellularity, 28.6%, lymphocytic rich, 3.5%, lymphocytic depletion 5.1%, nodular lymphocytic predominant, 7.5% and unclassifiable, 10.5%. Fifty percent of patients were stage III/IV and 46% had B symptoms. Clinical characteristics were not significantly different between patients treated with ABVD, ABV and COP/ABV (Table 1). Treatment options were varied according to doctors' preference, hospital strategy and the reimbursement policy. ABVD was the most common prescribed regimen (32.7%); the other options included ABV (29.2 %), COP/ABV (13.1 %), CHOP (2.8 %) and radiotherapy only (1.6%). With a median follow-up of 32 months, patients treated with ABVD had superior 5 year overall survival rate over those treated with ABV (86.6% vs 47.7%, p < 0.001) and COP/ABV (86.6% vs 62.7%, p < 0.01)(Figure 1). Discrepancies in survivals could be demonstrated both in patients with limited as well as in advanced diseases (Figure 2).

CONCLUSION: ABVD yielded superior survival compared to ABV and COP/ABV. Dacarbazine is therefore an essential drug for the treatment of HL in developing countries.

Figure 1. Figure 1. Figure 2. Figure 2.

Disclosures

Khuhapinant: Roche: Honoraria.
Follicular Lymphoma B Cells Generate Functional Regulatory T Cells Via ICOS/ICOSL Pathway and Are Inhibited By Intratumoral Tregs
Authors: Olive Daniel , LE Suong Khieu , Thibult marie-Laure , gondois-Rey Francoise , Granjeaud Samuel , Fest Thierry , Imene Azzaoui , Bouabdallah Reda , Broussais Florence , Xerri Luc , Leroux Dominique , Caux Christophe , Caux Christine
Published: November 2019

The prognosis of follicular lymphoma (FL) patients is suspected to be influenced by tumor infiltrating regulatory T cells (Tregs). The mechanism of Tregs enrichment in FL and their impact on malignant FL B cells remains to be elucidated. We analyzed 46 fresh lymph node biopsy samples including FL (n = 20), diffuse large B cell lymphoma (DLBCL; n = 10), classical Hodgkin lymphoma (cHL; n = 9) and reactive lymphadenitis (rL; n = 7). Using multicolor flow cytometry and cell sorting, we observed an accumulation of CD25high CD127low/- Tregs in FL tissues. These Tregs comprised activated ICOS+ Tregs which were able to suppress not only conventional T cells, but also FL B cells. These FL B cells were able to express ICOSL in vitro and to generate CD25high Foxp3high Tregs expressing ICOS. Tregs generation was associated with ICOS/ICOSL engagement and was abrogated by antagonist anti-ICOS and anti-ICOSL antibodies. Interaction between Tregs and FL B cells resulted in ICOSL downregulation on FL B cells. These data highlight the role of Tregs in FL pathogenesis and suggest that targeting the ICOS/ICOSL pathway may be a promising immunotherapy in FL

Disclosures

No relevant conflicts of interest to declare.
Prevalence and Prognostic Impact of CD30 and EBV Co-Expression in Korean Patients with EBV-Positive DLBCL of the Elderly
Authors: Hong Jung Yong , Park Keon Woo , Ko Young Hyeh , Kim Seok Jin , Kim Won Seog
Published: November 2019

The prognostic impact of EBV-positivity in EBV-positive DLBCL of the elderly is controversial. Previous Asian studies have shown consistent poor prognostic impact of EBV-positivity. Recent Western study has shown no significant prognostic impact of EBV-positivity alone. However, the study found that CD30 and EBV co-expression is associated with an adverse outcome and suggested CD30 co-expression might be associated with different prognostic impact of EBV-positivity between Asian and Western populations.

We retrospectively analyzed 26 Korean patients with EBV-positive DLBCL of the elderly diagnosed at Samsung Medical Center and had adequate amount and quality of paraffin-embedded biopsy specimen for additional immunostaining of CD30. EBV RNA was detected by an ISH technique and a positive reaction was defined as more than 20% of examined cells showing nuclear positivity. Cell of origin was assigned using Hans immunohistochemical algorithm.

Prevalence of CD30 co-expression in EBV-positive DLBCL of the elderly was 65.4% (17/26) and 50.0% (13/26) using a >0% cut-off and a >30% cut-off, respectively. There was no statistically significant difference of survival between cases with CD30 co-expression and case without CD30 co-expression. However, the cases with CD30 co-expression showed a modest trend of worse overall survival (OS) (median OS, 12.5 months [95%CI 1.5-23.4] versus not reached, P = 0.279) and progression-free survival (PFS) (median PFS, 7.4 [95%CI 5.7-9.1] versus 34.2 months [95%CI 0.0-89.1], P = 0.430) using >0% cut-off. Among 17 patients with CD30 and EBV co-expression, most of patients (88%, 15/17) showed non-germinal center B-cell like (GCB) DLBCL.

Substantial proportion of Korean patients with EBV-positive DLBCL of the elderly showed CD30 co-expression. CD30 co-expression in EBV-positive DLBCL of the elderly showed a modest trend of an adverse outcomes without statistical significance. Further validation study with larger population is warranted. Considering previously known contradictory trend of better outcome in CD30 positive cases with EBV-negative and GCB type DLBCL, we also need to investigate the possible distinct pathobiology of CD30 positivity in EBV-positive DLBCL of the elderly.

Disclosures

No relevant conflicts of interest to declare.
Anaplastic Large Cell Lymphoma of the Breast: Incidence, Patterns of Care, and Outcome in the US Population
Authors: Chalasani Pratima , Go Ronald S. , Parsons Benjamin , Al-Hamadani Mohammed
Published: November 2019

Background:

Anaplastic large cell lymphoma (ALCL) of the breast is a recently described clinicopathologic entity, almost always associated with breast implants, and does not overexpress the anaplastic lymphoma kinase (ALK) protein. Using the two largest US cancer databases, we describe its incidence, patterns of care, and clinical outcome.

Methods:

Incidence rate was calculated using 2000-2011 data from the Surveillance, Epidemiology, and End Results (SEER) Program and age-adjusted to the US 2000 standard population. For patient level data, we used the National Cancer Data Base (NCDB) Participant User File. We included all adult patients diagnosed from 1998-2011 with ALCL (ICD-O-3: 99714) who had breast as the primary site.

Results:

The incidence of breast ALCL was 0.001 per million. We identified 61 cases from NCDB. All of the patients were females and they were predominantly White (88.5%). The median age at diagnosis was 64 years (range, 18-90). The stage distribution was I (45.9%), II (21.3%), III (0%), IV (9.8%), and unknown (21.3%). The pattern of initial treatment was surgery alone (13.1%), chemotherapy alone (32.7%), chemotherapy and radiation (19.7%), surgery and chemotherapy (4.9%), surgery and radiation (4.9%), and tri-modality treatment (13.1%). In 7 patients (11.5%), no treatment was given. Two patients (1.6%) received stem cell transplant (1 autologous, 1 allogeneic) as part of initial treatment, one after chemotherapy and another one after surgery and chemotherapy. Overall survival data was available for patients diagnosed in 1998-2006 (29 patients). The median overall survival was 149.8 months (range, 2.6-157.9).

Conclusions:

ALCL of the breast is very rare. It presents mainly as early stage disease with very good prognosis. The patterns of care varied, although most patients received chemotherapy as part of the initial treatment.

Table 1. Treatment type received Frequency Percent Cumulative Frequency Cumulative Percent Chemo only 20 32.79 20 32.79 Surgery only 8 13.11 28 45.90 Chemo, Radiation, No surgery 12 19.67 40 65.57 Chemo, Surgery, No radiation 3 4.92 43 70.49 Radiation, Surgery, No Chemo 3 4.92 46 75.41 All three combined 8 13.11 54 88.52 Nothing 7 11.48 61 100.00

Disclosures

Parsons: Celgene: Speakers Bureau.
Lymphocyte/Monocyte Ratio (LMR) at Diagnosis Is Predictive of Survival with Post Transplant Lymphoproliferative Disease (PTLD) after Renal Transplant: An Analysis of 96 Patients of a Brazilian Cohort
Authors: Perini Guilherme Fleury , Cavalcanti Egyla M , Garibaldi Junior Joao , Penna Adriana M. Damasco , Pestana Jose Osmar Medina , Bordin Jose Orlando , Zampieri Filho Jose Pedro , Baiocchi Otavio C. G.
Published: November 2019

Introduction: Post-Transplant Lymphoproliferative Disease (PTLD) is a well-recognized group of lymphoid and or plasmacytic proliferations that occur following both solid organ (SOT) and allogenic hematopoietic stem cell transplantation (HSCT) as a result of immunossupression. A continuum of disease has been described, including early lesions, polymorphic PTLD and monomorphic PTLD. Epstein-bar virus (EBV) is considered the most important causative factor, with EBV positivity observed within up 90% of tumor lymphocytes. There is paucity of prognostic factors in PTLD, but studies have shown that IPI, Performance status and number of disease sites are related to mortality. LMR has shown to be prognostic in different lymphomas, including Hodgkin's Lymphoma (HL), Diffuse Large B-cell lymphomas among others.

Objectives: To assess the role of lymphocyte/monocyte ratio at diagnosis in predicting outcome and survival in PTLD patients.

Patient and Methods: This is a retrospective study conducted by the Universidade Federal de São Paulo, São Paulo, Brazil. Only confirmed cases of PTLD, diagnosed from 2001 to 2015, with clinical, epidemiological and laboratorial parameters available were included in this study. Event was defined as treatment related mortality, progression (defined as time for initiation of salvage therapy) or relapse. Patients with conflicted data or loss of follow up were excluded from the analysis.

Results: A total of 98 patients were diagnosed with PTLD in the study period. Two patients were excluded from the analysis due to conflict clinical data. Median age at diagnosis was 41.4 years (range from 4-74), with a 0,6:1 Female:Male ratio. Median time of transplant to PTLD was 56 months (range 0-967). Twenty-six patients (27%) received anti-thymocyte globulin (ATG), and the most common immunossupressive regimens consisted of cyclosporine or tacrolimus associated with prednisone and azathioprine (68,7%). Monomorphic PTLD was observed in 78% of patients, and two patients presented with HL. With a median follow up of 33.3 months, 35% of patients died. In a univariate analysis, the number of extranodal sites and LMR were associated with worst survival (p=0.026 and p=0.004).

Conclusions: PTLD is a known complication of immunossupresion, usually related to EBV. In our cohort of 96 kidney recipients with PTLD, LMR and number of extranodal sites was associated with inferior outcome.

Disclosures

No relevant conflicts of interest to declare.
The Prognostic Significance of 13q Deletion with Different Size in Patients with B-CLPD
Authors: Xiong Wenjie , Li Heng , Yi Shuhua , Li Zengjun , Liu Wei , Lv Rui , Zou Dehui , Qiu Lugui
Published: November 2019

Background: Deletion of 13q14 is the most frequent abnormality in chronic lymphocytic leukemia (CLL). Deletion at of 13q with vary sizes have been described, documenting different involved genes with different deletion area. But the prognostic significance of different deletion size is not well explored and this interesting phenomenon has not been described in other B-cell chronic lymphoproliferative disorders (B-CLPD).

Patients and Methods: Interphase fluorescence in situ hybridization (FISH) using locus-specific probes (RB1, D13S25) was performed to detect 13q14 deletions on 344 untreated cases with B-CLPD, including 304 CLL patients and 166 other B-CLPD patients. Different 13q14 deletion area was delineated by RB1 and D13S25.

Results: (a) Totally, 145 of 304 CLL patients (47.9%) had 13q14 deletion, which was significantly higher than it in other B-CLPD (25/167, 15.0%; p=.000). 77 of 304 CLL patients (25.3%) and 14 of 166 other B-CLPD patients (8.4%) had RB1 deletion. RB1 deletion always accompanied with D13S25 deletion in both CLL and other B-CLPD, which indicated large area deletion in 13q14. But some patients had only D13S25 deletion without RB1 deletion, indicating small area deletion in 13q14. Of the 145 CLL cases with 13q14 deletion, 73(50.3%) had small deletion and 72 (49.7%) with large deletion. The incidence of small and large deletion in 21 other B-CLPD with 13q14 deletion were in 12(48.0%) and 113(52.0%) patients respectively.

(b) In CLL cases, the time to first therapy in the cases with 13q14 deletion was 25 months longer than 16 months in the cases without 13q14 deletion (P=0.035). The progression-free survival (PFS) in cases with 13q14 deletion was also significantly longer (P=0.023) than those without the aberration. In other B-CLPD cases, the PFS and OS in cases with 13q14 deletion was significantly shorter (P=0.030) than those without the aberration. Moreover, large deletions (from RB1 to D13S25) were associated with a significantly shorter (p< 0.05) TFT as well as overall survival (57.5 vs. 135.0 months, p=.075)compared with small deletions.

Conclusions: The deletion of 13q can influence clinical outcome of BCLPD and vary in size both in CLL and in other BCLPDs. The deletion size displayed as a poor prognostic factor in other BCLPD cases, which implies the usage of both 13q probes is proposed.

Disclosures

No relevant conflicts of interest to declare.
Efficacy of Vindesine + Dexamethasone (VDS+Dexa.) in Elderly Patients with Relapsed or Refractory Malignant Lymphoma
Authors: Yuasa Hiromi , Wakita Hisashi , Matsuura Yasuhiro , Inagaki Shunichirou , Arai Hironori , Utsu Yoshikazu , Masuda Shinichi , Aotsuka Nobuyuki , Sunami Shosuke
Published: November 2019

Background; The number of non-Hodgkin's lymphoma (NHL) patients in the elderly has been increasing due to the long-lived society. It is not rare to treat elderly NHL patients aged over 85 years old. Individualizing the doses of cancer chemotherapy agents and progress in supportive therapy has improved the prognosis for elderly patients with NHL. Prolonged hospitalization elderly patients have adverse effects, which include dementia, difficulty in walking, and depression. Optimal treatment for these patients is unknown. Patients and Methods; In our hospital between April 2012 and December 2014, we treated 19 elderly patients (85 and over 85 years) with NHL as outpatients with VDS 3mg (day 1) and Dexa. (day 1-4) which repeated every 3 weeks for as long as possible. Results; Complete remission was achieved in 5 patients and partial remission in 6; the median duration of survival was 14 months. Adverse effects included leukopenia in 1 patient (<1,000 cells/μl), thrombocytopenia in 1patient (<5×104 cells/μl). Conclusion; VDS+Dexa. is useful in elderly patients because of its low hematological and non-hematological toxicity. These results indicate that VDS+Dexa. is effective and safe for the treatment of NHL in elderly patients. This outpatient chemotherapy caused no serious adverse reactions.

Disclosures

No relevant conflicts of interest to declare.
Successful Experience of Treatment of Angioimmunoblastic T-Cell Lymphoma By Prolonged Therapy
Authors: Chernova Natalia G. , Vinogradova Yulia E. , Sidorova Yulia V. , Kovrigina Alla M. , Gabeeva Nelly G. , Zvonkov Eugene E. , Parovichnikova Elena N. , Savchenko Valeri G.
Published: November 2019

Introduction: Angioimmunoblastic T-cell lymphoma (AITL) is a rare tumor derived from follicular T-helper and characterized by low response rate and relapsing course to standard therapy. CHOP-like therapy shows 70-80 % of overall response rate, though 2-year progression-free survival is observed only in 30-40 % of patients. Unsatisfactory results of treatment angioimmunoblastic T-cell lymphoma by ÑÍÎÐ-like therapy require searching new approaches to therapy.

Aim: To define a rational approach to treatment of angioimmunoblastic T-cell lymphoma.

Patients and methods: Ñlinical outcomes of 23 patients with newly diagnosed AITL treated between June 2002 and July 2015 we retrospectively analyzed. The diagnosis was verified according to WHO classification.

Results: Among the 23 patients 14 men and 9 women were included with a median age of 63 (range, 29-81). Twenty-two of 23 patients had Ann Arbor stage IV. Bone marrow involvement was detected in 21 of 23 cases by histological investigation. Nine of 23 patients received CHOP-like therapy; other 14 patients were treated prolonged therapy. Eight (89%) of 9 patients after CHOP-like therapy had treatment failure. Time to disease progression ranged from 1 to 7 months (median 3). Fourteen of 23 patients were treated by prolonged therapy consisting of low dose cytotoxic drugs weekly and supporting the therapy by interferon-A and thalidomide. Supporting therapy has administrated after achieving complete remission and was continued in 2 years. Prolonged therapy wasn't finished in 3 patients with severe infections and somatic pathology. Eleven (79%) of 14 patients treated by prolonged therapy achieved complete remission, treatment was finished only in 5 of 11 patients; follow-up varies from 3 to 67 months, a median 29. Univariate analysis of progression-free survival curves showed that prolonged therapy is more effective than CHOP-like therapy (p < 0.01) (Fig.1). Prolonged therapy was performed in 4 of 8 patients with progression disease after CHOP-like therapy; 2 patients achieved a complete remission, continuing 41 and 77 months.

Conclusion: Although, this was a small retrospective study, the results show that efficiency of prolonged therapy is better than CHOP-like therapy in patients withangioimmunoblastic T-cell lymphoma. Prolonged therapy allows preventing early treatment failures.

Figure 1. Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Late Cardiotoxicity of Intensive Modified Program NHL-BFM-90 in Adult Poor Prognosis Patients with Diffuse Large B-Cell Lymphoma
Authors: Dorokhina Elena , Magomedova Aminat Umaraskhabovna , Shevelev Alexander , Kulikov Sergei M , Gitis Michael , Vedernikov Alexey , Vedernikov Alexey , Vorobiev Andrey , Kravchenko Sergey , Savchenko Valeri G
Published: November 2019

Aim. To evaluate late cardiotoxicity of intensive modified program NHL-BFM-90 (mNHL-BFM-90) in adult poor-prognosis patients with diffuse large B-cell lymphoma (DLBCL).

Materials and methods. The data of ECG and echocardiography of 40 patients DLBCL wich received mNHL-BFM-90 protocol in the Hematology Reserch Center in the period from 2002 to 2009 years was analyzed . Group consisted of 20 men, 20 women, aged from 31 to 76 years, median age - 56.5 years at the time of the survey, the median period after therapy- 6 years. Individual cumulative dose of doxorubicin - 150 to 300 mg / m2. Comparison group included 19 patients, 8 men and 11 women, aged from 39 to 78 years, mediana- 70 years at the time of the survey, who received therapy to CHOP / R-CHOP. The median period after therapy - 5.5 years. Individual cumulative dose of doxorubicin - 200 - 400mg / m2. ECG and echocardiography studies were performed before and after 5 or more years after the end of chemotherapy. Clinical cardiomyopathy was defined by the presence of clinical signs of congestive heart failure. Subclinical cardiomyopathy was defined by the presence of characteristic changes by echocardiography and ECG study without clinical signs of CHF.

Results. From 40 patients DLBCL wich received mNHL-BFM-90, signs of subclinical cardiomyopathy were detected in 24 (60%) patients, clinical heart failure was not defined in any patient. In comparison group signs of subclinical cardiomyopathy were detected in 14 (74%) patients. Clinical heart failure was not defined in any case. Summar index of cardiotoxicity was significantly dependent on age (p=0.03), the presence of cardiac disease in anamnesis (p=0.3), and significantly higher (p = 0.05) in the group of patients after CHOP / R-CHOP therapy. A risk of subclinical cardiomyopathy revealing was significantly higher in group with cardiac disease in anamnesis (p = 0.05).

Conclusion. Late cardiotoxicity of high-dose program mNHL-BFM 90 does not exceed the toxicity of standard CHOP / R-CHOP. Patients with ischemic heart disease, hypertension, diabetes in anamnesis had the most severe sings of subclinical cardiomyopathy after chemotherapy according to examination data. Clinically significant heart failure in any patient was identified.

Disclosures

No relevant conflicts of interest to declare.
Autologous Stem Cell Transplantation (AutoSCT) in First Remission of Follicular Lymphoma (FL) "Save" Patients (pts) with Poor Prognosis. Results of the First Russian Prospective Study
Authors: Nesterova Ekaterina , Kravchenko Sergey , Baryakh Elena , Misyurina Anna , Mangasarova Yana , Vorobyev Vladimir , Gaponova Tatyana , Gemdzhian Eduard , Vorobyev Andrey , Savchenko Valeri
Published: November 2019

Background: FL is one of the most frequent types of lymphomas: it consists about 22% of non-Hodgkin lymphomas in Russia. 20 % of pts have an aggressive tumor after standard therapy.

In FL pts with unfavorable prognosis (FLIPI-1 III-IV, fast growth of tumor mass, B-symptoms, third cytological grade of tumor, bulky, absence of tumor response after first line R-CHOP-21 therapy) autoSCT allows to increase the overall (OS) and progression-free survival (PFS).

Aim: To estimate an efficacy of HDT (high dose therapy) with following autoSCT in front-line therapy of FL pts with factors of poor prognosis.

Patients and Methods: Since 2000 to 2015 years in National Research Center for Hematology were treated 39 FL pts with poor prognosis. All pts received R-CHOP-21 induction therapy. In 24 patients (62%) who hadn't partial response or poor prognosis patients with partial response after 4-6 courses autoSCT was performed. This group consists of 17 males and 7 females with median age 46 years old (31-68). Majority of pts (23/24) had IV stage of disease according to Ann Arbor. In 16/24 (66%) cases pts had bulky disease. 3/24 pts had phenomenon of leukaemization. Basing on FLIPI-1 criteria all pts were divided into 3 groups: low risk - 5/24, intermediate - 3/24, high - 16/24. B-symptoms were in majority of cases (18/24). 18/24 pts were diagnosed with I-II cytological grade of FL, 6/24 - III A/B. Basing on tumor growth pattern there was following ratio: nodular tumor growth revealed in 6/24 pts, nodular-diffuse tumor growth - in 15/24, diffuse tumor growth - in 3/24. Immunochemical analysis of serum and urine proteins was performed in 20/24 cases, among them increased level of serum β2-microglobulin was diagnosed in 10/20 pts. Increased activity of lactate dehydrogenase (LDH) was revealed in 16/24 pts. Bone marrow involvement at the time of disease onset was diagnosed in 18/24 pts.

Results: After induction R-CHOP-21 chemotherapy pts underwent HDT which included two courses R-DHAP, following high-dose cyclophosphamide with rituximab, then high dose etoposide with rituximab. As condition regimen R-BEAM was performed. All 24 FL pts, who underwent autoSCT, continue to be in complete remission. Median follow-up was 31 months (7-178).

Conclusions: Preliminary results of the first prospective study of intensive therapy of FL in Russia demonstrate that autoSCT in front line therapy provides to achieve a complete remission in pts with poor prognosis and allows to increase an overall and disease-free survival.

Disclosures

No relevant conflicts of interest to declare.