bluuebunny/crossref_metadata_2025 huggingface dataset:
Phase I/II Study of Escalating Doses of Idarubicin Orally Given with Oral Cyclophosphamide, Etoposide, Prednisolone and Intravenous Rituximab in Elderly Patients with Disseminated High Grade Non Hodgkin Lymphoma.
Authors: Fauveau Amélie , Lacotte-Thierry Laurence , Fleck Emmanuel , Colin Jean Yves , Guilhot François , Chabin Michèle , Langlois Alice , Guilhot Joëlle , Delwail Vincent
Published: October 2019

Abstract 3695

Poster Board III-631

Introduction

This study was performed to determine the maximum-tolerated dose (MTD) of oral idarubicin in combination with oral cyclophosphamide (CY), etoposide (ET), prednisolone (PRED) and intravenous (IV) rituximab (RI) administrated to previously untreated patients (pts) (60-80 years-old) with high grade stage III or IV non Hodgkin lymphoma. Secondary objectives were evaluation of toxicity and efficacy.

Patients and Methods

Idarubicin was administrated per os on day 1 with doses starting at 20 mg/m2 and escalated by 10 mg/m2 increments. Each idarubicin dose level included 3 to 6 pts. No intrapatient dose escalatation was allowed. Dose-limiting toxicity (DLT) was defined as grade 4 neutropenia lasting at least 4 days or grade 4 thrombopenia or more than grade 2 extra haematological toxicity. The MTD was defined as the dose level at which at least 2 of 3 or 3 of 6 pts experienced a DLT. CY and ET were given per os on day 1 to 3 at the fixed dose of 150 mg/m2 and 100 mg/m2 respectively. PRED was given at the fixed dose of 50 mg/m2 per os on day 1 to 5. RI was administrated IV on day 1 at the fixed dose of 375 mg/m2. G-CSF was systematically used from day 6 to haematological reconstitution. Eight cycles were planned at 3 week-intervals.

Results

Between October 2003 and August 2008, 19 immunocompetent pts [median age 70 (range 62-80)] with a new diagnosis of high grade non Hodgkin lymphoma received 128 cycles at 3 different idarubicin dose levels. No DLT was observed at dose level 1 (20 mg/m2), whereas 2 of 6 patients developed a DLT at dose level 2 (30 mg/m2). All 3 pts developed a DLT at level 3 (40 mg/m2) which was considered as MTD. Seven pts were enrolled at level 2 to confirm the results. Main toxicity results are summarized in the table below. No toxicity related mortality was observed. Two pts stopped treatment because of toxicity and 8 pts received a decreased dose. Overall response rate and complete response rate were 79% and 68% respectively. With a median follow-up of 22 months, the 2-year survival rates were 75% (95% CI: 45-90) for all pts and 58 % (95% CI: 21-83) for dose level 2 pts.

Conclusion

This first analysis clearly shows that the MTD of oral idarubicin administrated with CY, ET, PRED and RI, is 40 mg/ m2 on day 1 every 21-day cycle. The recommended dose study is ongoing. Overall survival data seems to be similar to previous publications. (Coiffier et al, NEJM 2002). Future development of subcutaneous anti-CD20 antibody could represent a special interest for this new oral chemotherapeutic protocol in order to improve the quality of life of elderly pts.

Disclosures:

No relevant conflicts of interest to declare.
The Histone Deacetylase Inhibitor MGCD0103 Down Regulates CD30, Activates NF-Kb, and Synergizes with Proteasome Inhibitors by HDAC6 Independent Mechanism in Hodgkin Lymphoma.
Authors: Buglio Daniela , Mamidipudi Vidya , Khaskhely Noor M , Gloghini Annunziata , Carbone Antonino , Heise Carla , Besterman Jeffrey M. , Martell Robert E , MacBeth Kyle J , Younes Anas
Published: October 2019

Abstract 3735

Poster Board III-671

Recent clinical trials demonstrated a promising clinical activity of the HDAC inhibitor MGCD0103 in heavily pretreated patients with relapsed HL. However, the mechanisms underlying MGCD0103 activity in HL remains unclear. Here we show that MGCD0103 preferentially inhibited HDACs1, 2 with no effect on HDAC6. Using three HL-derived cell lines, the IC50 for MGCD0103 after 72 hour of incubation was in the submicromolar concentrations, and was more effective than vorinostat in inducing cell death. Using gene expression profiling (GEP) studies, pathway PCR array, and western blot analysis, we identified several pathways that are modulated by MGCD0103. MGCD0103 downregulated the X-linked inhibitor of apoptosis (XIAP) protein, activated the intrinsic caspase pathway, and synergized with TRAIL agonistic antibodies. MGCD0103 downregulated CD30 mRNA and surface protein expression in a dose dependent manner, but had no significant effect on B cell-associated proteins, including CD19 and CD20, or on the costimulatory receptors CD40 and CD80. MGCD0103 induced TNF-alfa expression and secretion, which was associated with NF-kB activation, upregulation of the silencer of cytokine signaling (SOCS)-3, downregulated STAT6, and upregulation of STAT1 and 2. Selective inhibition of TNF-alfa expression by short interfering mRNA enhanced MGCD0103-induced cell death. Similarly, inhibition of NF-kB by proteasome inhibitors also potentiated the effect of MGCD0103, thus demonstrating synergy independent of HDAC6 inhibition. These findings demonstrate that MGCD0103 has a potent anti lymphoma activity by modulating the expression of a variety of survival proteins, and provides mechanistic rationale for combining class-I HDAC inhibitors with proteasome inhibitors and TRAIL.

Disclosures:

Mamidipudi: Celgene: Employment. Heise:Celgene Corporation: Employment, Equity Ownership. Besterman:Methylgene: Employment. Martell:Methylgene: Employment. MacBeth:Celgene Corporation: Employment, Equity Ownership. Younes:MethylGene: Honoraria, Research Funding.
Rituximab Does Not Improve Progression Free (PFS) or Overall Survival (OS) in Patients with Limited Stage Diffuse Large B Cell Lymphoma (DLBCL).
Authors: Hochberg Ephraim P. , Birrer Nicole , Toomey Christiana E. , Barnes Jeffrey A. , Lee Alfred Ian , LaCasce Ann , Abramson Jeremy S.
Published: October 2019

Abstract 3755

Poster Board III-691

Introduction

Diffuse Large B Cell Lymphoma (DLBCL) is the most common lymphoid malignancy accounting for approximately 33% of all newly diagnosed NHLs. Three randomized trials and multiple retrospective analyses have demonstrated both progression-free (PFS) and overall survival (OS) benefit to the addition of the anti-CD20 monoclonal antibody rituximab to anthracycline-containing chemotherapy in older advanced-stage patients, and in young low-risk patients with DLBCL. Approximately half of newly diagnosed patients with DLBCL present in limited stage and the benefit of rituximab containing chemotherapy regimens for these patients remains uncertain.

Methods

We used an IRB-approved clinicopathologic database, derived from comprehensive tumor registry data at the Massachusetts General Hospital, Dana Farber Cancer Institute and Brigham and Women's Hospital, to identify all patients 18 years or older diagnosed with limited stage DLBCL between 2000 and 2006. We included all patients treated with 3 or more cycles of anthracycline containing chemotherapy with curative intent. We excluded primary DLBCL of the CNS. We determined the impact of the use of rituximab on OS and PFS. PFS and OS were calculated from the date of initial diagnosis.

Results

A total of 138 patients met eligibility criteria and are included in the analysis. Median age was 51 years (range 18-89 years). 30% of patients were above 60 years of age, and less than 3% had an IPI score of 3 or higher. One hundred and six patients received CHOP + rituximab (RCHOP) and 32 received CHOP alone. Of the 106 patients receiving RCHOP, 48 were irradiated and 58 were not. Of the 32 patients receiving CHOP, 20 received radiation and 12 did not. At a median follow-up of 35 months (range 3-109 months), PFS and OS for the entire cohort are 86.2% and 90.6%, respectively. On univariate analysis of outcome, the addition of rituximab to CHOP did not improve PFS (81.3% vs. 87.7%,p=0.817, Logrank Test) or OS (84.4% vs. 92.5%, p=0.411, Logrank Test).

Conclusion

The outcome of an unselected series of patients with limited stage DLBCL is excellent. In this retrospective cohort of patients with limited stage DLBCL, the use of rituximab in conjunction with standard chemotherapy did not improve PFS or OS. The results we obtained are very similar to those reported by SWOG (Persky et al. JCO 2008). The overlapping confidence intervals for PFS and OS between SWOG 0014 and 8736 patients and our data suggest that a large multicenter trial will be needed to show a benefit of rituxan in this extremely good prognosis population.

Disclosures:

Hochberg: Genentech: Speakers Bureau; Biogen-Idec: Speakers Bureau; Enzon: Speakers Bureau; Amgen: Speakers Bureau.
Modulation of Cell Surface Protein Free Thiols; A Potential Novel Mechanism of Action of the Sesquiterpene Lactone Parthenolide in Non-Hodgkin's Lymphoma.
Authors: Skalska Jolanta , Brookes Paul S , Nadtochy S. M , Hilchey Shannon , Jordan Craig T. , Guzman Monica L. , Maggirwar Sanjay , Briehl Margaret M , Bernstein Steven H.
Published: October 2019

Abstract 3774

Poster Board III-710

Recent data supports the concept that redox regulation of cell surface protein thiols (i.e., exofacial thiols) effects critical cellular functions. We therefore hypothesized that redox-active anti-cancer therapeutics would modulate exofacial thiols, and that such modulation could be related to cell death mechanisms. To test this hypothesis, we used the sesquiterpene lactone parthenolide, a known anti-cancer agent. Parthenolide treatment results in a dose dependent decrease in mantle cell (MCL) and diffuse large cell lymphoma cell viability. Indeed, parthenolide decreases the level of free (ie. reduced) exofacial thiols on Granta MCL cells as assessed by flow cytometry. Parthenolide specifically modifies free thiols of surface proteins having molecular weights of ∼12 kd and ∼22 kd, as determined using a biotinylated thiol reactive reagent N-(biotinoyl)-N-(iodoacetyl) ethylendiamine (BIAM), which binds to free thiol groups and is detected by streptavidin-peroxidase staining of western blots. We further show that the central redox regulator thioredoxin is one of the surface protein thiol targets modified by parthenolide; specifically; (a) thioredoxin is detected on the Granta cell surface; (b) parthenolide directly interacts with free thiol groups on purified human thioredoxin, and; (c) parthenolide directly modifies Granta membrane associated thioredoxin, as determined with BIAM and neutravidin pull-down. To examine the functional consequences of parthenolide induced surface protein thiol modification, Granta cells were pretreated with the cell impermeable thiol antioxidant glutathione (GSH). Pretreatment with GSH inhibits the parthenolide induced; (a) decrease in exofacial free thiols; (b) modification of surface thioredoxin and; (c) Granta cell death. Pretreatment of Granta cells with GSH also inhibits parthenolide-mediated activation of JNK and inhibition of NFkB, two activities previously associated with the drug's anti-cancer activity. Based on these data, we postulate that at least one component of parthenolide's anti-lymphoma activity derives from its ability to modify the redox state of critical exofacial thiols. Indeed, to our knowledge, our data is the first to suggest that cancer cell exofacial thiols may be novel and important targets for cancer therapy.

Supported by an NCI SPORE grant in lymphoma 1P50 CA130805.

Disclosures:

Bernstein: millenium: Consultancy; genentech: Consultancy, Speakers Bureau; enzon: Consultancy.
Population Pharmacokinetics of Panobinostat (LBH589) in Patients with Advanced Solid Tumors and Hematologic Malignancies Following Intravenous and Oral Administration.
Authors: Savelieva Marina , Woo Margaret M. , Weber Harald A. , Hirawat Samit , Paul Sofia , Schindler Joanne , Fisch Roland
Published: October 2019

Abstract 3780

Poster Board III-716

Introduction

Panobinostat (LBH589), a hydroxamic acid derivative, is one of the most potent deacetylase inhibitors (DACi), with anti-tumor activity in a wide variety of preclinical xenograft models, and has shown promising clinical activity in patients with Hodgkin lymphoma and multiple myeloma. A population pharmacokinetic (PopPK) analysis was performed to characterize the extent and variability of panobinostat exposure in patients with advanced cancer and to evaluate the influence of selected covariates on exposure.

Patients and methods

Panobinostat concentrations in plasma were evaluated in 500 patients enrolled in five Phase I studies and four Phase II studies as asingle-agent administration. Patients received oral panobinostat at doses of 10–80 mg on Days 1, 3, and 5, weekly [QW], or every other week [QOW], or on Days 1 and 4 QW in three Phase I oral studies. Intravenous (i.v.) panobinostat was delivered at doses from 1.2 to 20 mg/m2 in 4 schedules (Days 1–7; or Days 1–3 QW or QOW; or once weekly for 3 weeks) in two Phase I i.v. studies. Panobinostat was also orally administered at 20 mg, on Days 1, 3 and 5 QW, in patients with cutaneous T-cell lymphoma, chronic myelogenous leukemia, and multiple myeloma in four Phase II oral studies. Panobinostat PopPK was characterized by a compartmental non-linear mixed-effects model.

Results

Panobinostat concentration–time data were well described by a linear 3-compartment model with first-order absorption. The model estimated the population median (CV%) of the total clearance (CL) to be 28.6 L/h (78%) and the volume of distribution (V) 27.2 L (54.8%). Absolute oral bioavailability is 30%. The relative bioavailabilities of oral formulations 1 (Phase II formulation) and 2 (Phase I formulation) are 17% and 23%, respectively. The analysis showed no effect on panobinostat clearance by gender, race, co-medications, disease state, or renal function as measured by plasma creatinine clearance. Body surface area (BSA 1.4–3.4 m2) and age (16–88 years) in the ranges studied were non-linear covariates for panobinostat clearance and volume.

Conclusions

The pharmacokinetics of panobinostat are linear and time independent. BSA may have clinical utility in predicting panobinostat clearance. The PopPK model generated from this study allows for future pharmacokinetic–pharmacodynamic correlation and application in optimizing the use of panobinostat.

Disclosures:

Savelieva Praz: Novartis: Employment. Woo:Novartis: Employment. Weber:Novartis Pharma AG: Employment. Hirawat:Novartis: Employment. Paul:Novartis: Employment. Schindler:Novartis: Employment. Fisch:Novartis : Employment.
The JNK Pathway Is a Significant Determinant of Sensitivity to DNA Damaging Agents in Pre-B ALL.
Authors: Saunders Philip O. , Bradstock Kenneth F , Bendall Linda J.
Published: October 2019

Abstract 3786

Poster Board III-722

The JNK pathway is reported to facilitate AP1 binding and promote apoptosis depending on cell type and environmental conditions. We have previously reported RAD001 (16μM) induces JNK pathway activation in pre-B ALL cells. We sought to evaluate the impact of changes in JNK pathway activation on pre-B ALL viability in vitro. Using JNK inhibitor SP600125 titrated to inhibit c-Jun activation, we determined that cell death in pre-B ALL cells treated with RAD001 (16μM) alone was not JNK dependent. In contrast, combining RAD001 (16μM) with DNA damaging agents significantly enhanced JNK dependent death. This difference indicates that additional factors, including genotoxic stress, are required for JNK activation to induce pre-B ALL cell death. The JNK pathway is reported to suppress transcriptional activation of key mediators of the DNA damage response. We observed that JNK activation in cells treated with RAD001 (16μM) and DNA damaging agents was associated with suppression of p53 and p21 relative to DNA damage alone. This result was supported by the observation of enhanced p53 and p21 expression in pre-B ALL cells treated with DNA damaging agents in the presence of the JNK inhibitor SP600125. Analysis of DNA content and proliferation antigen expression in pre-B ALL cells treated with RAD001 (16μM) and DNA damaging agents revealed JNK activation was associated with a significant increase in the proportion of cells in S phase, relative to DNA damage alone, which caused a G1 and G2 cell cycle arrest. Further evidence that the JNK pathway impacts on the DNA damage response was provided by the observation that pre-B ALL cells treated with DNA damaging agents and JNK inhibitor SP600125 demonstrated reduced PCNA expression at G1 and G2 and reduced expression of mitotic antigen phospho-Histone–H3. This is consistent with enhanced regulation at G1-S and G2-M checkpoints. The results indicate changes in JNK pathway activation impact on the cell cycle response to DNA damage. In conclusion we have identified that the JNK pathway has a significant impact on the sensitivity of pre-B ALL cells to DNA damaging agents. JNK activation in the presence of genotoxic stress significantly enhanced pre-B ALL cell death, associated with suppression of key mediators of the DNA damage response, p53 and p21. We found that changes in JNK activation altered the cell cycle response to DNA damage. Further study is required to determine if changes in cell cycle regulation in the presence of DNA damage is causal to JNK dependent cell death. Additional studies to identify intracellular signal pathways which facilitate JNK dependent cell death are warranted. Our observations suggest combining agents which induce JNK activation with conventional chemotherapy or selected novel agents has the potential to enhance clinical responses in pre-B ALL.

Disclosures:

No relevant conflicts of interest to declare.
The PI3K Inhibitor GDC-0941 Induces Growth Arrest and Apoptosis of Acute Myeloid Leukemia Cells.
Authors: Sun Laura , Berry Leanne , Du Changchun , Ma Xiaoju Max , Shi Lily , Medeiros Bruno C. , Ebens Allen J.
Published: October 2019

Abstract 3787

Poster Board III-723

Acute myeloid leukemia (AML) is a biologically and molecularly heterogeneous malignancy characterized by an accumulation of myeloid-lineage cells in the blood and bone marrow. Phosphatidylinositol 3' kinase (PI3K) -mediated signaling is frequently dysregulated in cancer and controls fundamental cellular functions such as cell migration, growth, survival and development of drug resistance in many cancers, including AML, and therefore represents an attractive therapeutic target, even against the backdrop of AML disease heterogeneity. In AML, for example, activating mutations in Ras and Flt3 are common and a number of autocrine and paracrine signaling mechanisms have been proposed as important disease mediators; all converge on PI3K as an important signal transduction node to provide growth- and survival-promoting effects. Here, we demonstrate that PI3K p110-a,b,d catalytic subunits are all prevalent across a panel of ∼30 cell lines and that the majority of cell lines as well as primary patient samples show evidence of PI3K pathway activation. We demonstrate in vitro with both cell lines and patient cell isolates, that a potent and selective pan-isoform PI3K inhibitor, GDC-0941, modulates pharmacodynamic markers such as p-Akt, p-4E-BP1, and p-FOXO-3a, and that treated cells show a G0/G1 cell-cycle arrest. A strong induction of apoptosis is observed by annexin/PI staining with upregulation of cleaved caspases and PARP. Preliminary observations suggest significant induction of p27 and Bim as direct transcriptional targets of FOXO-3a, as well as downregulation of c-Myc levels; these targets may represent key mediators of observed cellular effects and are the subject of ongoing work. Additional studies are currently underway to identify clinical standard-of-care agents and new molecular entities (pipeline NMEs) that combine favorably with PI3k-inhibition in vitro, and the results of several in-vivo xenograft studies will also be presented.

Disclosures:

Sun: Genentech Inc.: Employment. Berry:Genentech: Employment, Patents & Royalties. Du:Genentech, Inc.: Employment, Equity Ownership. Ma:Genentech: Employment, Patents & Royalties. Shi:Genentech: Employment, Patents & Royalties. Medeiros:Genentech: Research Funding. Ebens:Genentech, Inc.: Employment, Equity Ownership, Patents & Royalties.
Significant Association Between Promoter Polymorphism of the Erythropoietin Gene and Myelodysplastic Syndrome.
Authors: Ma Wanlong , Kantarjian Hagop , Zhang Ke , Zhang XI , Wang Xiuqiang , Zhang Zhong , Yeh Chen-Hsiung , Sferruzza Anthony , O'Brien Susan , Estey Elihu , Albitar Maher
Published: October 2019

Abstract 3825

Poster Board III-761

The risk of developing myelodysplastic syndrome (MDS) increases in the presence of a combination of genetic susceptibility factors and environmental carcinogenic factors. While numerous toxic agents have been described, genetic susceptibility factors for MDS are not well defined. A recent study showed that the TT genotype of the SNP rs1617640, located in the promoter of the eosinophile peroxidase (EPO) gene, is significantly associated with diabetic retinopathy and end-stage renal disease in patients with diabetes. Experimental models showed that the GG genotype is associated with several-fold lower erythropoietin expression than is the TT genotype. Since EPO plays a major role in the growth and differentiation of hematopoietic cells, we explored the association of the rs1617640 SNP genotype with MDS. We compared the EPO promoter SNP genotype in 187 MDS patients, 256 AML patients, and 95 normal control individuals. The GG genotype was significantly more common in patients with MDS (25.1%) than in normal control subjects (6.3%) and AML patients (12.5%) (P=0.0003); the GT genotype was present in 38.4% of MDS patients, 43.2% of AML patients, and 46.2% of normal control subjects. Combined analysis of data from MDS and control subjects indicated an elevated risk of MDS in individuals with the GG genotype relative to those with a non-GG genotype (odd ratio [OR] =4.98; 95% confidence interval [CI]=2.04-12.13). The GG (vs non-GG) genotype was also associated with elevated MDS risk when analysis was limited to the MDS and AML groups (OR =2.35; 95% CI=1.43-3.86). In contrast, analysis of AML and control group data showed a statistically insignificant elevated risk for AML in individuals with the GG genotype (OR=2.12; 95% CI=0.86-5.24). The odds of MDS and AML did not differ significantly with respect to GT (vs non-GT) or TT (vs non-TT) genotypes when compared with control subjects. Clinical and follow up data were available for 112 MDS patients and 186 AML patients. There was no correlation between EPO promoter genotype and response to therapy or overall survival in MDS or AML. In the MDS group, the GG genotype was significantly associated with shorter complete remission duration, as compared with the TT genotype (P=0.03). We found no correlation between EPO genotype and cytogenetic abnormalities, performance status, or other laboratory variables. In conclusion, the EPO promoter GG genotype shows strong association with MDS. Since MDS is well documented to develop after exposure to toxic agents, the clinical value of screening for the EPO promoter GG genotype in individuals at high risk due to exposure to toxic agents, then storing hematopoietic stem cells from those with the GG genotype for possible future use, should be investigated.

Disclosures:

No relevant conflicts of interest to declare.
Gene Expression Profiling Reveals Activation of Multiple Oncogenic Pathways and Over-Expression of Survivin in the Pathogenesis of Extranodal Nasal-Type NK/T Cell Lymphoma.
Authors: Chng Wee-Joo , Selvarajan Viknesvaran , Huang Gaofeng , Zhou Jianbiao , Law Mark , Feldman Andrew L , Manual Salto-Tellez , Ng Siok-Bian
Published: October 2019

Abstract 3931

Poster Board III-867

Extranodal nasal-type Natural Killer/T-cell lymphoma (NKTCL) is a distinct clinicopathologic entity most commonly affecting Asians and Central and South Americans, and characterized by a clonal proliferation of NK or T cells with a cytotoxic phenotype. There is a strong association with Epstein-Barr Virus. The tumor is aggressive with patient usually surviving short duration even with chemotherapy. We performed the first comprehensive genome-wide gene expression profiling (GEP) of extranodal nasal-type Natural Killer/T-cell lymphoma (NKTCL) using formalin-fixed paraffin embedded (FFPE) tissue (n=25) and NK cell lines (n=5) and compared the results to the GEP of normal NK cells using the Illumina DASL whole genome array, with the objective of understanding the oncogenic pathways involved in the pathogenesis of NKTCL and to identify potential therapeutic targets. Quantitative-PCR validation of the GEP findings revealed over-expression of candidate genes BIRC5 (survivin), EZH2 and STMN1 in NK cell lines compared to normal NK cells, consistent with the GEP data. We then extracted a list of genes that are differentially expressed between NKTCL and normal NK cells and tissue controls. We than subjected this list of genes to pathway and network analysis using Metacore. This revealed a significant enrichment for cell cycle related genes and pathways. Furthermore, the network analysis results demonstrated a pro-proliferative and anti-apoptotic phenotype in NKTCL characterized by activation of Myc and nuclear factor kappa B (NF-KB), and deregulation of p53. This was further corroborated using immunohistochemistry on tissue microarray of NKTCL (n=33, including all cases with GEP performed). We observed a significant percentage of NKTCL showing overexpression for c-Myc (45.4%), p53 (87.9%) and NF-KB p50 (67.7%) on immunohistochemistry. Notably, overexpression of survivin was observed in 97% of the cases. Based on our findings, we propose a model of NKTCL pathogenesis where deregulation of p53 together with activation of MYC and NF-KB, possibly driven by EBV LMP-1, result in the cumulative upregulation of survivin. When KHYG and NKYS cell lines were treated with a compound IDR E804 which inhibited survivin, there is significant inhibition of cell growth as assess by MTS assay and induction of apoptosis as measured using Annexin V staining by flow cytometry. This suggests that compounds inhibiting survivin may be a potentially useful novel therapeutic approach in NKTCL.

Disclosures:

No relevant conflicts of interest to declare.
Prognostic Impact of Mid-Treatment PET in Patients with Diffuse Large B-Cell Lymphoma Who Achieved Metabolic CR at Post-Treatment PET
Authors: Yoo Changhoon , Kim Jeong-Eun , Sohn Byeong Seok , Yoon Dok Hyun , Kim Shin , Lee Dae Ho , Kim Sang We , Lee Jung Shin , Suh Cheolwon
Published: October 2019

Abstract 3949

Poster Board III-885

Background

Mid-treatment positron emission tomography (PET) has been found to predict clinical outcomes in patients with aggressive non-Hodgkin's lymphoma. Currently, risk-adapted therapy based on mid-treatment PET has been widely evaluated. This study was intended to assess the prognostic value of mid-treatment PET in patients who achieved metabolic complete response (mCR) at post-treatment PET.

Methods

From February 2002 to March 2009, total 130 patients in whom post-treatment PET showed mCR were included in this study. We performed retrospective analysis of progression-free survival (PFS) and overall survival (OS) according to the results of mid-treatment PET.

Results

Median age (range) was 51 (16-85) years old, and 70 (54%) patients were male. International Prognostic Index (IPI) was low (0-2) in 91 (70%) patients and high (3-5) in 39 (30%) patients. As a front-line chemotherapy, most frequently administered regimen was R-CHOP (76%). Eighty-seven (67%) patients were mCR, and 43 (33%) patients were metabolic partial response (mPR) in mid-treatment PET. With 24 months of median follow-up, 3 year-rates of PFS and OS in overall patients were 74% and 87%, respectively. Differences of survival outcomes between patients with mCR and mPR at mid-treatment PET were not statistically significant in terms of PFS (p=0.13) and OS (p=0.76).

Conclusions

In patients with metabolic CR at post-treatment PET, survival outcome was not influenced by the results of mid-treatment PET. Therefore, risk-adapted therapy solely based on mid-treatment PET might be inappropriate in the management of newly diagnosed DLBCL.

Disclosures:

No relevant conflicts of interest to declare.
Out of Hospital Treatment of Acute Pulmonary Embolism in Patients with a Low NT-ProBNP Level.
Authors: Agterof Mariette J , Schutgens Roger E.G. , Snijder Repke J , Epping G. , Peltenburg Henny G , Posthuma Ward , Hardeman J. A. , Van der Griend Rene , Koster Ted , Prins Martin , Biesma Douwe
Published: October 2019

Abstract 3996

Poster Board III-932

Background

NT-proBNP is emerging to be of great importance in risk assessment of patients with acute pulmonary embolism (PE), by identifying both high and low-risk patients. The aim of the present management study was to investigate the safety of out of hospital treatment of patients with acute PE based on a NT-proBNP < 500 pg/ml.

Methods

Hemodynamically stable outpatients with objectively diagnosed acute PE and a NT-proBNP level < 500 pg/ml, were considered eligible for participation in this multicenter prospective study. Patients were discharged immediately from the emergency room or within the first 24 hours of admission. The primary objective was the 10-days mortality rate. Secondary objectives were the incidence of re-admission to hospital due to PE or its treatment, the patient's satisfaction during the first ten days of treatment and the incidence of serious adverse events in a 3-month follow-up period.

Results

Approximately 40 percent of all patients presenting at the emergency room with acute PE fulfilled the inclusion criteria. In total, 152 patients (mean age 53.4 ± 14.3 years; 51% female) with PE were treated as outpatients. No deaths, major bleedings or recurrent venous thromboembolism occurred during the first 10 days or in a follow-up period of 3 months. Seven patients required readmission. Three patients were admitted because of complaints of their PE: two patients with anxiety and pain and one patient with dyspnoea and low oxygen saturation. There were no signs of progression of PE in these 3 patients. Four patients were re-admitted because of an illness unrelated to PE. According to the PSQ18 and anxiety score, patients were satisfied with their out of hospital treatment; they had no progression of anxiety during the first ten days of treatment.

Conclusion

Out of hospital treatment is safe in a group of low risk patients with non-massive PE, based on NT-proBNP levels < 500 pg/ml. Approximately 40 percent of PE-patients can be treated in an outpatient setting. Patients consider out of hospital treatment as comfortable and have no increase in anxiety scores.

Disclosures:

No relevant conflicts of interest to declare.
Fucosyltransferase VI Induces Platelet Activation: A Novel Property of a Plasma Glycosyltransferase.
Authors: Navarro José-Tomás , Tawfiq Shwan , Wohlgemuth Roland , Hoffmeister Karin M. , Sackstein Robert
Published: October 2019

Abstract 4016

Poster Board III-952

A number of glycosyltransferases are present in human plasma with the α(1→3) fucosyltransferase, Fucosyltransferase VI (FTVI), having the highest plasma concentration. Notably, elevated plasma levels of FTVI are associated with a variety of cancers and correlate with tumor load/progression. The well-known association of neoplasia with thromboembolic complications prompted us to examine whether FTVI has direct effect(s) on platelet function. We obtained human platelets from blood of healthy donors and separated from platelet-rich plasma by differential centrifugation. Freshly isolated platelets (x108/ml) were stirred and exposed at 37°C to varying concentrations (20, 40, 60 and 80 mU/mL) of glycosyltransferases FTVI, β-1-4-galactosyltransferase-I (βGalT-I), or α,2-3-N-sialyltransferase (α2,3-N-ST), or to 1 U/mL thrombin. Platelet aggregation and activation was assessed by aggregometry (light transmission) or by flow cytometry of FSC/SSC characteristics and of surface expression of P-Selectin, respectively. FT-VI reproducibly induced platelet aggregation and activation, whereas other glycosyltransferases (β4GalT-I and α2,3-N-ST) had no effect on platelets. FTVI activation of platelets was concentration-dependent, and the aggregation curve for FTVI was one wave, similar to that for thrombin. FTVI-induced platelet activation was independent of catalytic conversion of surface glycans, but was inhibited by FTVI denaturation, indicating that FTVI-induced platelet activation is a lectin-mediated process. To determine the membrane target(s) mediating FTVI-induced platelet activation, biochemical studies were performed after catalytic exofucosylation of the platelet surface. Flow cytometry after platelet exofucosylation showed formation of the carbohydrate structure sLex, detected by the mAb Heca452, but no formation of Lex (CD15). Western blot showed that enforced fucosylation induced sLex on a single platelet surface protein, and further biochemical studies revealed that this protein is GPIbα. These findings unveil a previously unrecognized property of FTVI as an activator of platelets, mediated via a specific lectin/carbohydrate interaction on GP1ba, and offer novel perspectives on the pathobiology of tumor-associated thrombogenesis.

Disclosures:

No relevant conflicts of interest to declare.
Low-Dose Rituximab in Idiopathic Autoimmune Hemolytic Anemia.
Authors: Barcellini Wilma , Zaja Francesco , Zaninoni Anna , Imperiali Francesca Guia , Battista Marta , Filí Carla , Russo Domenico , Di Bona Eros , Consonni Dario , Zanella Alberto
Published: October 2019

Abstract 4027

Poster Board III-963

Background

Conventional therapy of warm autoimmune hemolytic anemia (WAIHA) include administration of corticosteroids and immunosuppressive agents, or splenectomy, whereas no effective treatment exists for cold hemagglutinin disease (CHD). A substantial proportion of patients with WAIHA do not respond to or relapse after corticosteroid therapy and may experience clinically relevant side effects. Favorable responses to rituximab at standard doses (375 mg/m2 weekly for 4-6 courses) have been reported in both WAIHA and CHD, idiopathic or secondary, as well as in other autoimmune diseases, such as rheumatoid arthritis and primary immune thrombocytopenia. Recently, low dose (LD) rituximab (100 mg fixed dose weekly for 4 courses) has been proven effective in patients with autoimmune cytopenias, particularly immune thrombocytopenia.

Aims

To evaluate the safety, activity and the duration of the response of LD rituximab associated with standard oral prednisone (PDN) as first line therapy in newly diagnosed WAIHA and CHD, and as second line therapy in WAIHA relapsed after standard oral PDN.

Methods

In this single-arm prospective pilot study, LD-rituximab was administered at 100 mg fixed dose weekly on days +7, +14, +21, +28 along with standard oral PDN (1 mg/kg/die p.o. from day +1 to + 30, followed by quick tapering: 10 mg/week until 0.5/mg/kg/die, then 5 mg/week until stop). Complete and partial initial responses (iCR and iPR)) were defined as Hb ≥ 12 g/dL and ≥ 10 g/dL at month +2 from the beginning of therapy, respectively; sustained response (SR) was defined as Hb ≥ 10 g/dL at month +6 and +12, in the absence of any treatment.

Results

Twelve patients (6 female, 6 male; median age 49 yrs, range 28-65) were enrolled. A iCR and iPR were observed in 6 and 4 out of 12 patients, respectively; the median Hb level increased from 9.9 g/dL (range 6.1-12.3) at enrolment to 12.3 g/dL (range 9.5-14.9) at month +2. A SR at month +6 was observed in 7 out of 7 evaluable patients, and in the first 2 patients enrolled at month +12. No side effects or serious adverse events were observed. For 9 relapsed AIHA patients laboratory data (Hb, LDH, reticulocytes) and steroid administration were available before LD rituximab treatment: a lower cumulative dose (roughly 50%) of steroid was administered to patients during LD rituximab study compared with previous therapy, without significant differences in the general trend of Hb, LDH, and reticulocytes.

Conclusions

These preliminary results seem to indicate that the addition of LD rituximab to standard corticosteroid therapy is a feasible and active treatment in AIHA. Data on SR are intriguing, particularly regarding the possible steroid sparing effect of LD rituximab, but need to be confirmed in larger survey and after longer follow up.

Disclosures:

No relevant conflicts of interest to declare.
Knockdown of Matriptase-2 Increases Hepcidin Expression in a BMP-Dependent Manner in Zebrafish Embryos.
Authors: Gibert Yann , Lattanzi Victoria J. , Fraenkel Paula G.
Published: October 2019

Abstract 4050

Poster Board III-985

Hepatocytes produce hepcidin, a transcriptionally regulated peptide hormone, in response to iron overload, resulting in decreased intestinal iron absorption. Hepcidin levels are inappropriately low in patients with thalassemia major and hemochromatosis. We have been developing the zebrafish embryo as a model to identify hepcidin regulators, which may be exploited to prevent and treat iron overload syndromes. Recently, it has been shown in mammalian models that the membrane-bound serine protease, matriptase-2 (also known as TMPRSS-6), inhibits hepcidin expression by cleaving membrane-bound hemojuvelin (hjv), a bone morphogenic protein (BMP) co-receptor, which is mutated in patients with juvenile hemochromatosis. Previously, we demonstrated that hepcidin expression increases in response to iron loading or activation of the BMP signaling pathway, but that hjv is not required for hepcidin expression in zebrafish embryos. We hypothesized that hepcidin expression in zebrafish embryos is normally hjv-independent, either because of low levels of hjv expression in the liver or because of the presence of matriptase-2. We used whole mount in situ hybridization, quantitative realtime RT-PCR, morpholino knockdowns and overexpression experiments to address these questions. As hjv was undetectable in the zebrafish embryonic liver by in situ hybridization, we used flow cytometry to sort zebrafish embryonic hepatocytes and assess hjv expression by RT-PCR. We found that hjv was weakly expressed in the embryonic liver, but strongly expressed in zebrafish adult liver. We overexpressed zebrafish hjv in human hepatocytes and found that it potentiated the effects of BMP6 to activate a human hepcidin promoter and a BMP response element. In contrast, overexpression of zebrafish hjv in zebrafish embryos failed to increase hepcidin expression. Knockdown of matriptase-2 in zebrafish embryos caused developmental delay and a smaller liver, but increased hepcidin expression, relative to liver size. Furthermore, treatment with the BMP inhibitor dorsomorphin abrogated the increased hepcidin expression associated with matriptase-2 knockdown. Experiments are underway to determine whether zebrafish hjv has little effect on hepcidin expression during embryonic development because of the activity of matriptase-2.

Disclosures:

No relevant conflicts of interest to declare.
Does Transcription of Remote α-Globin Regulatory Elements Affect Their Function?.
Authors: Kowalczyk Monika S , Hughes Jim R , Sharpe Jacqueline A , Brown Jill M , Buckle Veronica J , Wood William G , Higgs Douglas R
Published: October 2019

Abstract 4060

Poster Board III-995

Expression of many tissue and developmental stage specific genes is controlled by remote regulatory elements. The mammalian a-globin gene cluster includes four previously characterised remote regulatory elements (MCS-R1 to R4) upstream of the a-globin genes. Naturally occurring deletions and experiments in transgenic mice, have shown that one or more of these elements are necessary for fully regulated expression of the a-like globin genes. At present it is unclear exactly how these elements interact with the a-globin genes to enhance their expression, although ultimately we know that they physically interact with the promoters forming a chromatin loop. Of interest, three of the MCS-R sequences are located within introns of a widely expressed gene C16orf35, which lies adjacent to the a-globin locus and we now know that such complex, intermingled arrangements are common in the mammalian genome. We have asked two questions: can the MCS-R elements be transcribed while activating the globin genes, and does transcription of the MCS-R elements play any role in their activation?

We have analysed nascent transcription of C16orf35 and a-globin, and shown that both genes are simultaneously transcribed in a high proportion of erythroid cells. Thus it appears that the MCS-R elements can interact with the globin genes while being transcribed. We next asked whether transcription of the C16orf35 gene (through the upstream regulatory elements) is necessary for their activation. The promoter region and a putative erythroid-specific promoter of the C16orf35 gene were deleted (singly and in combination) by homologous recombination in mice. Provisional analysis has shown no evidence of a-thalassaemia in these mice, suggesting that activation of the MCS-R elements occurs independently of their transcription. In addition to their importance in globin gene regulation, these findings have general implications for the relationship between structure and function in the mammalian genome and the mechanisms by which long-range elements interact with their cognate promoters.

Disclosures:

No relevant conflicts of interest to declare.
In Vitro Generation of Cytotoxic T Lymphocytes against MAGE A1 and MAGE A3 Derived From Healthy Donors.
Authors: Bao Lei , Stamer Mindy M , Dunham Kimberly , Krishnadas Deepa Kolaseri , Lucas Kenneth G
Published: October 2019

Abstract 4079

Poster Board III-1014

MAGE A1 and MAGE A3 are cancer testis antigens that are expressed on a number of malignant tumor cells, but not by normal cells, except for male germ cells which lack HLA expression. Therefore, MAGE cytotoxic T lymphocytes are strictly tumor-specific. Adoptive transfer of antigen specific cytotoxic T lymphocytes (CTL) provides immediate graft-versus tumor effects while minimizing risk for graft-versus-host disease. The aim of the current study was to find ideal conditions for expansion of CTL targeting tumor-associated antigens from peripheral blood mononuclear cells (PBMCs) of healthy donors to be used in allogenic cell therapy. In this study we investigated the ability to generate MAGE A1 and MAGE A3 specific cytotoxic T cells using autologous dendritic cells (DC) loaded with MAGE A1 and MAGE A3 overlapping peptides. CTL lines specific for MAGE A1 and MAGE A3 were established by stimulating CD8 T cells from healthy donors with autologous dendritic cells loaded with MAGE A1 or MAGE A3 overlapping pooled peptides in round-bottomed, 96-well plates. CD8+ T cells were restimulated with the same ratio of peptide pulsed DC on days 7 and 14 in the presence of IL-2 (50 U/ml), IL-7 and IL-15 (5 ng/ml). These microcultures were screened 10 days after the third stimulation for their capacity to produce interferon-gamma (IFN-gamma) when stimulated with autologous EBV-transformed B lymphocytes (BLCL) transduced with lentivirus(LV) encoding MAGE A1 or MAGE A3 and autologous BLCL transduced with LV encoding GFP. MAGE A1 and MAGE-A3 specific IFN-gamma producing cells were rapidly expanded in OKT3 and IL2. The specificity of the rapidly expanded MAGE A1 and MAGE A3 specific T cells was confirmed by IFN-gamma production as measured by intracellular cytokine staining and ELISA as well as antigen specific cytotoxicity by a standard 51chromium (51Cr) release assay. We successfully generated MAGE A1 and MAGE A3 specific CTL lines from healthy donors using this method. Specific CTL lines showed cytotoxicity in vitro not only to target cells pulsed with MAGE A1 or MAGE A3 peptides but also to target cells transduced with LV-MAGE A1 or LV-MAGE A3. Specific cytolytic activity was accompanied by IFN-gamma secretion. These data indicate that tumor antigen specific CTL can be expanded using overlapping peptides regardless of an individual's HLA specificity. The ability to generate tumor specific CTL from donors of various HLA backgrounds provide a rationale for utilizing MAGE A1 and MAGE A3 overlapping peptides for expansion of antigen specific T cells for adoptive T-cell therapy against MAGE A1 or MAGE A3 expressing tumors.

Disclosures:

No relevant conflicts of interest to declare.
Adult Philadelphia-Positive Acute Lymphoblastic Leukemia (Ph + ALL) Treated at Diagnosis with Imatinib Followed by 2nd Line TKI (Imatinib or Dasatinib) and Stem Cell Transplantation.
Authors: Carella A. M. , Catania G. , Pica G. , Beltrami G. , Nati S. , Vimercati R. , Congiu A. , Rossi E. , Spriano M.
Published: October 2019

Abstract 4109

Imatinib has become an integral part of front-line therapy for Ph+ ALL, with remission rate exceeding 90% irrespective of whether Imatinib is given alone or combined with chemotherapy. Treatment outcome with Imatinib-based regimens has improved survival compared with historic controls, but most patients who do not undergo allogeneic stem cell transplantation (SCT) eventually relapse. Second generation TKI, e.g. Nilotinib and Dasatinib, show activity against most of the BCR-ABL tyrosine kinase domain mutations involved in acquired Imatinib resistance, but clinical benefit is generally short lived. In the attempt to improve the prognosis of these patients, we started a new intensive protocol consisting of induction phase with Imatinib (600 mg po, daily) in combination with conventional chemotherapy; moreover, two intrathecal methotrexate will be administered at the day +8 and +22. As known, this approach will determine an high number of complete hematologycal (CHR) and cytogenetic remissions (CcyR). Soon after, Imatinib will be discontinued and a 2nd TKI (Nilotinib or Dasatinib) will be offered to the patients. The idea is to reduce/eliminate as soon as possible the mutations generating under Imatinib. The primary objective of the study will be to assess the activity of sequential TKI to induce molecular remission; secondary objectives will be DFS, relapse rate, OS. This protocol will be designed for patients >18 years. Recruitment started in June 2008. Minimal residual disease will be investigate by RQ-PCR at day +30, +60, +90, +120, +150 and so on. The protocol has been designed for 20 patients. Three evaluable patients entered until now this trial; the median age was 47 years (range, 39-64 years). All 3 patients achieved CHR and CCyR at a median of 26 days (range, 18-31 days) and 46 days (range, 39-59 days), respectively. Monitoring RQ-PCR, a marked clearance of leukemic cells was found at d+60; when CCyR was achieve, Imatinib was discontinued and 2nd TKI was started: 2 patients received Dasatinib 100 mg po daily and 1 patient Nilotinib 800 mg po twice daily. Between d+100 and d+120, RQ-PCR became negative in the peripheral blood of all patients. One patient with an HLA+ sibling donor is waiting to receive an allograft; the other 2 patients, who did not find an HLA+ donor, received an autograft with molecular negative PBPC. The conditioning regimen consisted of the BU-Cy regimen. Both patients are alive and in CMR at 3 and 10 months after autografting. More patients and more time will confirm the efficacy of this new approach.

Disclosures:

No relevant conflicts of interest to declare.