bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rituximab Plus Bendamustin (R-B) Treatment In Patients with Aggressive Large B-Cell Lymphoma (LBCL): Response and Tolerability - a Single Institution Experience
Authors: Horn Julia , Kleber Martina , Kohlweyer Ulrike , Hieke Stefanie , Herzog Regina , Schmitt-Gräff Annette , Engelhardt Monika 
Published: October 2019

Abstract
Abstract 4900

Introduction:
Clinical studies have shown that Rituximab plus Bendamustin (R-B) in indolent lymphoma results in favourable responses, progression free survival (PFS) and lower toxicity as compared to R-CHOP. The aim of this analysis was to characterize response and tolerability of R-B in patients with LBCL, who were not qualifying for R-CHOP due to age, comorbidity and/or prior pretreatment (including anthracyclines).

Methods:
We retrospectively identified consecutive patients with LBCL receiving at least two cycles of R-B in our department between 2003 and 2010 using our electronic tumor documentation system. Patient characteristics, response to R-B, and toxicity were assessed.

Results:
We identified 9 caucasian patients (5 females, 4 males) with LBCL; their median age was 71 years (range; 51–82). Two presented with stage I/II, seven with stage III/IV disease at initial diagnosis and before R-B. Six patients had a low or intermediate IPI and three were high risk. Four patients received R-B as first-line therapy, and five were treated for relapsed or refractory disease.
Main determinants for the R-B-selection were contraindications for anthracyclines in five patients and advanced age and/or poor performance status in four patients.
A median of four R-B-cycles were applied (range; 2–6). Response with achievement of CR and PR was observed in 6/9 (CR: 2, PR: 4), two achieved SD. Only one pt showed PD after four R-B cycles. The response of R-B in first-line vs. relapsed appeared similar.
Of note, one female patient with secondary LCBL, after initial Hodgkin's lymphoma and C-MOPP chemotherapy (CTx) and mediastinal irradiation - with excellent response to R-B- failed to successfully mobilize PBSC thereafter. However, she was effectively mobilized with R-Ara-C-thiotepa (peripheral blood CD34+ cells were 5.82 vs. 54/μl, obtaining no vs. 6.72 × 106 CD34+ cells/kg KG via leukapheresis, respectively). Clinical tolerance of R-B in all patients was excellent in a total of 31 R-B-cycles, only two major CTC-events occurred: one infection (CTC grade III) and one thromboembolism (grade IV). Median PFS and overall Survival (OS) were 16 (7- not reached) and 20 (11-21) months.

Conclusions:
If standard R-CHOP cannot be given due to age, comorbidity or CTx-contraindications (e.g. anthracyclines), R-B may represent an effective treatment in LBCL. Larger cohorts and prospective clinical trials are needed to confirm these promising results. Currently, patients with grade III/IV follicular lymphoma are additionally evaluated for response and tolerability under R-B, also being presented at the meeting.

Disclosures:
No relevant conflicts of interest to declare.
PI3KCA and PIM Inhibitors in Peripheral T-Cell Lymphomas
Authors: Martin-Sanchez Esperanza , Rodriguez-Pinilla Socorro M. , Lombardia Luis , Sanchez-Beato Margarita , Dominguez-Gonzalez Beatriz , Romero Diana , Gomez-Abad Cristina I. , Vargiu PierFrancesco , Rodriguez Maria E. , Mollejo Manuela , Alves Javier , Wozniak Magdalena B. , Bischoff James R. , Piris Miguel A. 
Published: October 2019

Abstract
Abstract 4917
T-cell lymphomas (TCL) are a heterogeneous group of aggressive malignancies lacking specific and efficient therapy. Unfortunately, there are neither animal models nor representative cell lines for most TCL types, making functional and pharmacogenomics studies even more difficult. PI3K and PIM are kinases involved in cell proliferation, frequently altered in human cancer that seems to play a critical role in T-cell development and activation. Genomic studies have identified PIK3CD subunit to be significantly associated with in activation of CD40, NF-kB and TCR-pathways. The aim of this project is to determine the efficiency of PI3K inhibitors (PI3Ki) and PIM inhibitors (PIMi) in TCL, looking for biomarkers of their mechanism of action and to identify markers that could identify responders from non-responders.
Twenty PTCL and seven reactive lymph nodes were studied using gene expression microarrays. We performed an in silico analysis using the Connectivity Map program to identify drugs that could potentially reverse PTCL gene expression signature. Among them, several PI3K/mTOR inhibitors were found. A panel of 6 TCL cell lines belonging to different TCL subgroups were treated with 3 PI3Ki (LY294002, ETP-45658, GDC-0941) and one PIMi (ETP-39010). Functional studies were also done to establish the role of each of the targeted genes.
In vitro studies showed that PI3Ki induced G1 cell cycle arrest in all cell lines, and apoptosis in a portion of them, in a time/dose-dependent manner. We also observed a decrease in the levels of pAKT(S473), pGSK3B(S9) and p-p70S6K(T389) after treatment.
In addition, both the analysis of the PTCL gene expression signature as well as western blot studies on TCL cell lines has shown overexpression of PIM family genes, A decrease in cell viability, and a strong induction of apoptosis in all cell lines was seen after PIM inhibition, without cell cycle arrest. Several diagnostic and pharmacodynamic biomarkers of PIMi have been identified at the mRNA and protein level in both cell lines In conclusion, our results indicate that PI3Ki and PIMi are effective therapeutic approaches for TCLs, identifying potential markers for patient's stratification and pharmacodynamic assessment.

Disclosures:
No relevant conflicts of interest to declare.
Combinatorial Effects of CD30 Ligand Stimulation or Inhibitor of Apoptosis (IAP) Antagonist Exposure with Conventional Chemotherapy on CD30 Positive Malignancies
Authors: Walkovich Kelly J , Liu Xuwen , McCollom Anne L , Duckett Colin S 
Published: October 2019

Abstract
Abstract 4926
CD30 is a member of the tumor necrosis factor (TNF) receptor family that is normally found on the cell surface of a small subset of activated lymphocytes but is overexpressed on the surface of anaplastic large cell lymphoma (ALCL) and Hodgkins lymphoma (HL) cells. Although many drugs exist that treat lymphomas by triggering the intrinsic cell death pathway, current chemotherapeutic regimens are limited by unwanted side effects, including secondary malignancies that limit event-free survival. The tumor-restricted overexpression of CD30 makes it an attractive target for therapeutic intervention. Depending on the cellular context, CD30 stimulation has been linked to cell death, cell cycle arrest, or paradoxically, proliferation. In ALCL tumor cell lines, CD30 stimulation activates both the canonical and noncanonical NF-kB pathways while in HL tumor cell lines, CD30 stimulation only slightly enhances NF-kB activity above constitutive levels, implying a role for NF-kB in determining the sensitivity or resistance of lymphoma cells to CD30-induced apoptosis. In addition, IAP antagonists, small synthetic compounds that mimic the structure of the second mitochondrial activator of caspase (Smac) and target IAP molecules that affect the activation of the non-canonical NF-kB pathway, induce apoptosis and/or sensitize cells to death via secondary signals such as TNF. This suggests that the modulation of IAP levels, and consequently regulation of the non-canonical NF-kB pathways, may also have a role in determining tumor cell death. Using representative ALCL and HL tumor cell lines, we have found that CD30 stimulation via its physiologically ligand in combination with standard chemotherapeutic agents results in increased efficacy in tumor cell death in the majority of ALCL cell lines but not HL cell lines. Similarly, IAP antagonists in combination with standard chemotherapeutic agents also resulted in enhanced tumor cell death in most ALCL but not HL cell lines. This augmentation of tumor cell death suggests that CD30-induced apoptosis and IAP antagonist-induced killing may have important consequences in the clinical treatment of CD30 positive malignancies. Currently, we are further investigating the role of both CD30 stimulation via its physiological ligand and IAP antagonists in impacting the activation of the canonical and noncanonical NF-kB pathways alone and in combination with currently utilized chemotherapeutic agents to modulate the apoptotic threshold in CD30 positive lymphoma cells.

Disclosures:
No relevant conflicts of interest to declare.
Heart Function In Systemic and Localized AL Amyloidosis: Role of NT-ProBNP and MPC-1
Authors: Saltarelli Francesca , Moscetti Alessandro , Bruno Guglielmo , Monarca Bruno , Salerno Gerardo , Corsetti Maria Teresa , Cardelli Patrizia , Ferranti Giovanna , La Verde Giacinto 
Published: October 2019

Abstract
Abstract 5006
In AL amyloidosis typical sites of amyloid buildup are heart, skin, gastrointestinal tract, liver, kidneys, and blood vessels. To evaluate the heart involvement in systemic and localized amyloidosis proBNP, peptide (NT-proBNP; 76 amino acids) and MPC-1 were investigated. NT-proBNP have been described as useful marker for the diagnosis heart disease, and its plasma concentrations correlate with the functional classification of patients according to the New York Heart Association (NYHA). MCP-1 is a chemokine that activates mononuclear phagocytes by promoting leukocyte–endothelium binding and migration to sites of inflammation. The MCP-1 levels seem to be related to the severity of cardiac alteration, as demonstrated by the coronary angiogram. NT-proBNP and MPC-1 serum levels were performed in systemic or localized AL amyloidosis to evaluate if there was a difference in the heart involvement. Blood samples were collected from 8 patients with systemic amyloidosis and from 4 patients with localized amyloidosis. To analyze the results of NT-proBNP and MPC-1, Mann-Whitney test was performed. NT-proBNP serum values were significantly (p=0.007) increased in systemic disease. Also, MPC-1 serum levels were significantly (p=0.004) higher in the patients with systemic disease (350.52±58.70 pg/ml) if compared to the group of localized amyloidosis (147.82±26.03 pg/ml). On the basis of our results, the heart seem to be functionally more involved in AL systemic amyloidosis than in localized disease, as demonstrated by the higher NT-proBNP and MPC-1 serum values.

Disclosures:
No relevant conflicts of interest to declare.
Proteasome Inhibitor Bortezomib Disrupts Tumor Necrosis Factor-Related Apoptosis-Inducing ligand (TRAIL) Expression and Natural Killer (NK) Cell Killing of TRAIL Receptor-Positive Multiple Myeloma Cells
Authors: Feng Xiaoli , Yan Jie , Wang Yibiao , Zierath Juleen R. , Nordenskjöld Magnus , Henter Jan-Inge , Fadeel Bengt , Zheng Chengyun 
Published: October 2019

Abstract
Abstract 5021
Bortezomib, a potent 26S proteasome inhibitor, is approved for the treatment of multiple myeloma (MM) and clinical trials are under way to evaluate its efficacy in other malignant diseases. However, cytotoxic effects of bortezomib on immune-competent cells have also been observed. The aim of this study was to investigate the putative effects of Bortezomib on activated human natural killer (NK) cell function. In this project, primary lymphocytes or NK cells were isolated or purified from peripheral blood mononuclear cells of healthy donors. Expression of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) by interleukin (IL)-2-activated NK cells was quantified by flow cytometry and real time-PCR method. NK cell degranulation activity and cytotoxicity against MM cell lines were evaluated by CD107a surface translocation and 51Cr release assays, respectively. Our results demonstrate that bortezomib markedly downregulated cell surface expression of TRAIL on primary human IL-2-activated NK cells in a dose-dependent manner. Moreover, bortezomib reduced TRAIL mRNA expression in these cells, suggesting that bortezomib regulates TRAIL expression of activated NK cells at the transcriptional level. Interestingly, pharmacological inhibition of the transcription factor NF-κB also profoundly decreased TRAIL expression both at protein and mRNA levels, indicating a novel role of NF-κB in the regulation of TRAIL expression in activated human NK cells. Furthermore, perforin-independent killing of the human MM cell lines was significantly suppressed following bortezomib treatment. In addition, blocking cell surface-bound TRAIL with a TRAIL antibody impaired lymphokine-activated killer (LAK) cell-mediated lysis of the TRAIL-sensitive MM cell line, RPMI8226. Our study demonstrated that bortezomib decreases TRAIL expression by IL-2 activated NK cells in a dose-dependent manner and disrupts TRAIL-mediated killing of TRAIL-sensitive myeloma cells, suggesting that Bortezomib may potentially hamper NK-dependent immunosurveillance against tumors in patients treated with this drug.

Disclosures:
No relevant conflicts of interest to declare.
Inhibition of JAK2V617F Kinase Potently Induce Apoptosis In HEL Cells
Authors: Shen xu-Liang , Wei Wu , Xu Hong-liang , Qin Xiao-qi , Fangping Chen 
Published: October 2019

Abstract
Abstract 5062
Myeloproliferative neoplasms (MPNs), formerly recognized as myeloproliferative disorders(MPDs), are a group of hematological conditions where a molecular defect in a multi-potent hematopoietic stem cell leads to increased production in one or more blood cell lineages. In early 2005, four different groups described an identical JAK2V617F mutation in exon 12 of JAK2 in a large number of patients with MPNs. Whereafter, it has been confirmed that the Gain-of-function mutations of JAK2V617F play crucial roles in the development of MPN. However, how this activated singaling pathway leads to overproduction of blood cells are not fully understood. Our experiment was designed and performed to address one aspect of this issue. Here we report that AG490, a JAK2 kinase inhibitor, is potently inducing apoptosis in HEL cells. HEL cell line, which harbor a homozygous JAK2V617F mutation confirmed by Allele-specific polymerase chain reactions (AS-PCR), restriction enzyme digestion and DNA sequencing, was selected as a cell model. HEL cells were treated with AG490 at different concentrations and for different time length. The Flow cytometry(FCM) with AnnexinV-FITC/PI double staining and genome DNA electrophoresis were performed to determine the apoptosis rate of the HEL cell, respectively; RT-PCR was performed to analyze the effect of AG490 on the expression of Bcl-x mRNA. The FITC-AnnexinV/PI double staining demonstrated that the apoptosis rate of HEL cell increased obviously in a time (F=267.226, P<0.0001)and dose(F=343.77, P<0.0001) dependent manner after treated by AG490. Similar results were observed by DNA ladder. The results of RT-PCR demonstrated that when treated by AG490, the mRNA of Bcl-xL gene was down-regulated obviously(Bcl-xL mRNA was the main species of Bcl-x found), and the down-regulation was strongly time(F=26.402, P<0.0001) and dose(F=39.247, P<0.0001) dependent. Thus, our results suggest that AG490 can induce apoptosis in HEL cells obviously, and JAK2V617F might inhibit cell apoptosis through upregulation of Bcl-xL.

Disclosures:
No relevant conflicts of interest to declare.
T(8;22)(q24;q11) In a Context of Complex Karyotype In Two Patients with B-Cell Lymphoma with Features Intermediate Between Burkitt Lymphoma and Diffuse Large B Cell Lymphoma B (LB/LDCGB)
Authors: Vallespi Teresa , Ortega Margarita , Jaramillo Anny , Lopez-Andreoni Laura , Martínez-Morgado Noemi , Sánchez Carmen , Bosch Francesc 
Published: October 2019

Abstract
Abstract 5093

Background:
In some cases, it is difficult to distinguish between the diagnosis of Burkitt lymphoma (LB) and Diffuse Large Cell Lymphoma B (DLCL-B) with MYC gene rearrangement. The WHO classification distinguishes three clinical entities: LB, LB/DLCL-B, DLCL-B on the basis of morphology, immunophenotype and genetic characteristics of malignant cells. The MYC gene is often rearranged with the gene that encodes the heavy chains of immunoglobulins (IgG 14q32) and, less frequently, with the gene that encodes the light chains kappa (2p12) or lambda (22q11).

Objective:
Describe clinical and biological characteristics of two cases of lymphoma with variant t(8;22)(q24;q11) in a complex karyotype diagnosed BL/DLCL-B according to the WHO classification.

Methods and Patients:
The cytogenetic study was made in the bone marrow (aspirate or biopsy). Leukocytes were incubated during 24h in RPMI without mitogens. Rearrangements of C-MYC, BCL-2, BCL-6 and IGH were studied for FISH techniques. The results were expressed according to ISCN 2009.

Case 1:
A man of 46 years who was admitted with symptoms of spinal cord compression with fever and weight loss. During admission were detected anti-HIV antibodies. In the biopsy were observed large cells with irregular nuclei, with clefts and some multilobulated. Imunohistochemistry: CD10 +, CD20 +, CD79 +, bcl2 + and BCL6 +. The Ki67/MIB1 was 95%. Cytogenetic study of bone cylinder showed a complex karyotype: 49, XY, +1, del (1) (p22), der (2), der (3), +7, t (8;22)(q24;q11), +10, del(10)(q24q26)x2 [3]. The FISH study demonstrated the rearrangement of C-MYC and the absence of BCL2 and BCL6 rearrangement.

Case 2:
A man of 67 year old was admitted for epigastric pain, nausea, vomiting and weight loss. The aspirated cells were of medium and large size, irregularly shaped nuclei, some creases and other poly-lobed with prominent nucleoli. Was detected by immunohistochemistry positivity: CD10, CD20, CD79 and BCL6 Ki67/MIB1 presented a 90%. Cytogenetic study of the bone marrow showed a complex karyotype: 49, XY, t(1;11)(q21;p13),+7, t(7;13) q32;q12), t (8;22)(q24;q11),+12,+19[40]. The FISH study demonstrated presence of the rearrangement of C-MYC and BCL2 and BCL6 rearrangement absence.

Conclusions:
- Both cases are representative of the clinical entity BL// DLCL-B recently recognized by the WHO: anisocitosicas cells with a nucleus of irregular or poly-lobed, with Ki67/MIB1 <90% and C-MYC rearranged with a complex karyotype. - Both cases showed the t(8;22)(q24;q11) rare variant of Burkitt lymphoma and a trisomy 7, trisomy usually observed in DLCL-B.

Disclosures:
No relevant conflicts of interest to declare.
Upregulation of MAGE-A1, MAGE-A3, NY-ESO-1 and MHC Molecules on Neuroblastoma Cells Following Exposure to Decitabine and Interferon-γ
Authors: Bao Lei , Dunham Kimberly , Lucas Kenneth 
Published: October 2019

Abstract
Abstract 5179
MAGE-A1, MAGE-A3, and NY-ESO-1 are cancer-testis (CT) antigens expressed on a number of malignant solid tumors, including neuroblastoma (NB), but many tumor cell lines down-regulate the expression of CT antigens as well as major histocompatibility (MHC) antigens, precluding recognition by antigen specific T cells. If expression of cancer antigens as well as MHC molecules could be enhanced pharmacologically, the efficacy of CT antigen specific immunotherapy could be improved. DNA methylation is an epigenetic mechanism used by cells to decrease the expression of several genes, including tumor suppressor genes. Decitabine (5-aza-2′-deoxycytidine, DAC) is a potent inhibitor of DNA methylation, and the doses associated with the demethylating action of this drug are much lower than those required for cytotoxicit. We have demonstrated that the expression of MAGE-A1, MAGE-A3, and NY-ESO-1 can be upregulated on neuroblastoma cells following exposure to the DAC. When these cell lines are cultured in the presence of DAC at concentrations similar to levels achieved with clinical administration, we were able to achieve expression of NY-ESO-1, MAGE-A1, or MAGE-A3 in 11/11 neuroblastoma cell lines. Concentrations of 1uM DAC were associated with strongest expression of these antigens, which was optimal after 72 hours of drug exposure. Culture of NB cell lines with IFN-γ was also associated with an increased expression of either MHC Class I or II by cytofluorometry, and Class I upregulated in 10/11 of these cell lines, although to varying degrees. MAGE-A1, MAGE-A3, and NY-ESO-1 specific CTL were cultured from volunteer donors by stimulating peripheral blood mononuclear cells with dendritic cells pulsed with overlapping peptide mixes derived from full length proteins, and these CTL lysed HLA partially matched, DAC treated neuroblastoma as well as glioblastoma cell lines. These studies confirm that DAC and IFN-γ can be used to increase the expression of CT antigens and MHC molecules on neuroblastoma cells, and pre-treatment with these agents makes tumor cell lines more susceptible to CTL mediated killing.

Disclosures:
No relevant conflicts of interest to declare.
Frontline High Dose Chemotherapy Supported by Autologous PBSCT Might Improve Survival In Elderly Patients (≥60 Years) with High Risk Diffuse Large B-Cell Lymphoma (DLBCL)
Authors: Ivanov Vadim , Coso Diane , Rey Jerome , Aurran-Schleinitz Therese , Stoppa Anne-Marie , de Colella Jean-Marc Schiano , Blaise Didier , Bouabdallah Reda 
Published: October 2019

Abstract
Abstract 5190
Limited data is available concerning feasibility and efficacy of high dose therapy (HDT) supported by autologous PBSCT in elderly patients with non-Hodgkin lymphoma (NHL). In young patients with poor prognostic features intensification supported by PBSCT as a part of first-line treatment suggests survival benefit. It is not clear if the same strategy is applicable to the older patients.
The Institute Paoli-Calmettes database was reviewed for all DLBCL patients who received BEAM followed by PBSCT in patients >=60 years old between January 1998 and January 2010 (13 years). All patients were HIV-negative and received BEAM intensification as a part of front-line treatment. All of them were in a complete response after CHOP or R-CHOP induction prior to autograft. Thirty six auto-transplanted patients were identified (median age 63 y, range 60–68). This cohort was compared with closely matched group of 43 patients of same age range, who received first-line CHOP or R-CHOP regimen without intensification in the same 13-year interval. Only patients in a complete response after first line were included.
As frontline autoPBSCT was performed in high-risk patients, the group without HDT was naturally privileged in the terms of Ann-Arbor stage and aaIPI index. There was significant difference in the localised vs disseminated disease (stage I-II: 53,4% in no-HDT vs 14% in HDT group, p=0,00025) and aaIPI (0-1: 72% in no-HDT vs 28% in HDT, p=0,000086) between the two groups. Factors evaluated included treatment-related mortality (TRM), overall survival (OS) and event-free survival (EFS). There were no transplant-related deaths in the HDT group.
The estimated 5-year OS was 81,3% (95% CI 62,5-91,9 %) and 10-year OS was 65% (95%CI 34,2-86,9%) for HDT group compared to 91,5% (95% CI 77,5-97,1%) and 58,4% (95% CI 20–88,8%) in the no-HDT group (p=NS). There were 8 events (6 relapses and 2 secondary malignancy deaths) in the HDT group and 11 events (9 relapses and 2 secondary malignancy deaths) in no-HDT (5-year EFS 77,6% (95% CI 60,3-88,8%) vs 78,3% (95%CI 62,3-88,8%), p=NS)).
We conclude that front-line autologous PBSCT with BEAM conditioning can be safely performed in patients aged 60 years or above with DLBCL after CHOP of R-CHOP induction. There was no difference in OS and EFS between cohorts with and without intensification even if the auto-transplantation procedure was reserved for the high risk patients only. The first-line HDT with autologous PBSCT in older patients with high-risk IPI score might improve survival in this group and produce results similar to those in the low-risk group.

Disclosures:
No relevant conflicts of interest to declare.