bluuebunny/crossref_metadata_2025_split huggingface dataset:
Heparanase Activation of the Coagulation System in a Mice Sepsis Model
Authors: Henig Israel , Axelman Elena , Brenner Benjamin , Vlodavsky Israel , Nadir Yona 
Published: October 2019

Abstract
Abstract 378

Background:
Heparanase that is abundant in platelets and neutrophils is implicated in cell invasion, tumor metastasis and angiogenesis. Recently, we demonstrated that heparanase is directly involved in the regulation of the hemostatic system. Heparanase was shown to form a complex and enhance tissue factor (TF) activity, resulting in increased factor Xa production (Nadir et al, Haematologica, 2010). In the present study, the effect of heparanase on sepsis was investigated. Methods: ICR mice, heparanase knock-out mice and heparanase over-expression mice were studied. Sepsis was induced by intra-peritoneal injection of lipopolysaccharides (LPS). Four hours after injection, blood was obtained via temple plexus puncture followed by mice sacrificing. Levels of thrombin-antithrombin (TAT), D-Dimer and IL-6 were tested using ELISA. Plasma factor VII levels were evaluated using the Western blot analysis and fibrinogen levels in the lung, kidney and spleen were estimated by immunostaining. Results: Intra-peritoneal injection of heparanase (0.5μg/mg) compared to PBS injection, increased the TAT (p<0.05) and D-Dimer (p<0.05) levels, which were similar to the ones induced by LPS (5μg/mg), although the IL-6 levels did not rise and mice did not show signs of illness. Moreover, heparanase injection half an hour following LPS administration resulted in reduced levels of TAT, D-Dimer, fibrinogen and of factor VII (p<0.001), increased levels of IL-1 (p<0.001) and profuse blood leak from puncture, compatible with severe disseminated intravascular coagulation (DIC). Similarly, injection of LPS to heparanase knock-out mice led to lower levels of TAT (p<0.001), D-Dimer (p<0.001) and IL-6 (p<0.001) compared to control mice, while injection of LPS to heparanase over-expression mice resulted in DIC. Conclusions: Heparanase is found to induce activation of the coagulation system without inflammatory response. It also contributes to laboratory enhancement of sepsis resulting in DIC. Inhibitors of heparanase may potentially attenuate morbidity and mortality in sepsis.

Disclosures:
No relevant conflicts of interest to declare.
Outcome of Complex Variant Philadelphia Positive Chromosome Translocations in the Era of Tyrosine Kinase Inhibitors,
Authors: Shah Bijal D. , Zhang Ling , Bulkeley William , Moscinski Lynn , Papenhausen Peter , Pinilla-Ibarz Javier 
Published: October 2019

Abstract
Abstract 3788

Background:
It is important to prospectively identify patients with CML who might not respond to conventional tyrosine kinase inhibitory therapy. Cytogenetic variants of the Philadelphia (cvPh) chromosome can be observed in 5–8% of patients diagnosed with Chronic Myelogenous Leukemia (CML), and usually involve at least one or more chromosome other than 9 and 22. Specifically, cvPh refers to patients with (V;9;22), where “V” indicates one or more additional translocation partners. Previous data suggest that patients with cvPh may respond poorly to therapy (Gorusu et al, 2007), however, many were treated prior to routine use of imatinib, nilotinib, or dasatinib.

Methods:
We conducted a retrospective review of 338 patients with CML treated at Moffitt Cancer Center to investigate the presence of cvPh and the outcome of these patients under treatment with TKI therapy.

Results:
Of 338 patients we identified 12 patients with cvPh (3.5%). Demographic data show male to female ratio of 5:7, and median age 50 (35–76). Two were in accelerated phase at the time of diagnosis, the remaining were in chronic phase. Baseline cytogenetics from time of diagnosis were not available in six patients. Median time to development of cvPh was 10 months (0–74). Two did not have clearly identified cvPh at diagnosis but both had complex karyotype. Eleven patients were treated with imatinib, among whom only one has shown a complete cytogenetic response (CCR). Two others continue on imatinib with unknown response status. Four patients were treated with dasatinib, including one in the frontline setting. No CCR's have been documented in this group, with two patients pending reevaluation. Seven patients were treated with nilotinib after failing a prior TKI, of whom only one has shown a CCR, with response evaluation pending in two patients. Two patients have undergone allogeneic stem cell transplant, with resolution of disease, including the nilotinib treated patient in CCR above. In this series of 12 patients, only three have shown resolution of variant translocation with a median followup of 45 months. All patients are alive at the last follow up.

Conclusion:
In this retrospective review, patients with cvPh appear to have a more resistant genomic type, and seems to be a bad prognostic factor for response to TKI therapy.

Disclosures:
No relevant conflicts of interest to declare.
Ponatinib Is Active Against the CUX1-FGFR1 Fusion Kinase and Against Imatinib Resistance Mutations of the FIP1L1-PDGFRα Fusion Kinase and of KIT,
Authors: Lierman Els , Smits Sanne , Cools Jan , Dewaele Barbara , Debiec-Rychter Maria , Vandenberghe Peter 
Published: October 2019

Abstract
Abstract 3848
Imatinib (IM) was initially developed as a small molecule inhibitor of the BCR-ABL1 kinase, but also potently inhibits other oncogenic kinases, such as PDGFRβ and PDGFRα fusion proteins. IM has revolutionized the treatment of chronic myeloid leukemia (CML) and other neoplasms, but the development of IM resistant mutations has emerged as an important problem, triggering a search for novel compounds that overcome resistance. A leading third generation candidate is ponatinib, a novel multikinase inhibitor with potent activity towards BCR-ABL1, KIT, FGFR1, PDGFRα and other kinases. Importantly, ponatinib also targets numerous IM resistant BCR-ABL1 kinase domain mutations including the panresistant T315I mutation.
We investigated the effect of this compound on IM resistant kinase mutations in lymphoid/myeloid neoplasms associated with eosinophilia and rearrangements of PDGFRα and FGFR1, as well as KIT associated malignancies. Ba/F3 cells were used expressing either FIP1L1-PDGFRα, the IM resistant FIP1L1-PDGFRα-T674I mutant, the panresistant FIP1L1-PDGFRα-D842V mutant, or the novel CUX1-FGFR1 fusion. In addition, several KIT mutants were investigated. The growth of FIP1L1-PDGFRα and IM resistant FIP1L1-PDGFRα-T674I mutant expressing cells was strongly inhibited by ponatinib with IC50 values of 0,6 nM and 9 nM respectively. Also the panresistant FIP1L1-PDGFRα-D842V mutant and the novel CUX1-FGFR1 fusion responded well to ponatinib treatment with 50% growth inhibition at 154 nM and 56 nM respectively. IL3-driven growth of Ba/F3 cells was resistant to ponatinib (IC50: 2 μM). Western blot analysis confirmed the direct effect of ponatinib on the auto-phosphorylation of the PDGFRα and FGFR1 fusion proteins, as well as on the downstream signaling protein STAT5. Finally, we investigated several KIT single and double mutants and preliminary data indicate an inhibitory effect of ponatinib towards several KIT mutants.
In conclusion, our results demonstrate the in vitro activity of ponatinib against IM resistant mutants of the FIP1L1-PDGFRα fusion kinase, against the CUX1-FGFR1 fusion kinase as well as against IM resistant KIT mutations. Our data indicate that ponatinib, which is currently under investigation in phase II clinical trials for IM resistant CML, may also be active against neoplasms driven by FGFR1, PDGFR or KIT kinase activity, and able to overcome IM resistance in these malignancies.

Disclosures:
No relevant conflicts of interest to declare.
Metascoring and Gene Expression Profiling in Clinical Routine in Multiple Myeloma,
Authors: Meißner Tobias , Seckinger Anja , Rème Thierry , Hielscher Thomas , Möhler Thomas , Neben Kai , Goldschmidt Hartmut , Klein Bernard , Hose Dirk 
Published: October 2019

Abstract
Abstract 3940

BACKGROUND.
Multiple myeloma is characterized by molecular heterogeneity transmitting to survival ranging from several months to over 15 years. Gene expression profiling allows assessment of biological entities, risk, and targets. Its translation into clinical routine alongside conventional prognostic factors has been prevented by lack of appropriated reporting tools and the integration with other prognostic factors into a single risk stratification (metascoring).

METHODS.
We present here a non-commercial open source software-framework developed in the open source language R (GEP-report) containing a graphic user interphase based on Gtk2. Affymetrix microarray raw-data and a documentation-by-value strategy to directly apply thresholds and grouping-algorithms from a reference cohort of 262 myeloma patients are used. Gene expression-based and conventional prognostic factors are integrated within one risk-stratification (HM-metascore) and the final report is created as a PDF-file.

RESULTS.
The GEP-report comprises i) quality control, ii) test of sample identity, iii) biological classifications of multiple myeloma, iv) risk stratification, v) assessment of target-genes, and vi) integration of expression-based and clinical risk factors within one metascore. This HM-metascore sums over the weighted factors gene-expression based risk-assessment (UAMS-, IFM-score), proliferation, ISS-stage, t(4;14), and expression of prognostic target-genes (AURKA, IGF1R) for which clinical grade inhibitors exist. It delineates three significantly different groups of 13.1, 72.1 and 14.7% of patients with a 6-year survival of 89.3, 60.6 and 18.6%, respectively.

CONCLUSION.
GEP-reporting allows prospective assessment of target gene expression and integration of current prognostic factors within one risk stratification (metascoring), being customizable regarding novel parameters or other cancer entities.

Disclosures:
No relevant conflicts of interest to declare.
Regulatory T Cells of G-CSF Mobilized Stem Cell Donors Qualify for Clinical Applications,
Authors: Ukena Sya N. , Velaga Sarvari , Lilia Goudeva , Ivanyi Philipp , Ganser Arnold , Franzke Anke 
Published: October 2019

Abstract
Abstract 4055
Recent clinical studies demonstrate the high potency of human regulatory T cells (Tregs) to control graft-versus-host disease following hematopoietic stem cell transplantation (SCT). Isolation of Tregs after recombinant G-CSF induced mobilization of stem cells would simplify the design of clinical trials in allogeneic SCT. However, there is growing evidence that G-CSF also exerts profound immune modulatory effects in the adaptive immune system. Therefore, we analyzed Tregs isolated by FACS based cell sorting from stem cell donors before (n=8) and after (n=13) G-CSF administration regarding their phenotype, stability, functional and expansion properties. Absolute CD4+ T cell (3.2 fold increase of mean after G-CSF; p<0.05) and CD4+CD25hi Treg cell numbers (4.1 fold increase of mean after G-CSF; p<0.01) were significantly increased after G-CSF administration. Analysis of the Foxp3 TSDR demethylation level displays a stable Foxp3 phenotype of G-CSF encountered Tregs (mean value 97.1% vs. 95.0 % after G-CSF administration). Moreover, the CD4+CD25hi Tregs of G-CSF treated SC donors suppress the proliferation of effector T cells with no significant differences to Tregs isolated from healthy donors before G-CSF treatment (mean values 42.1% vs. 49.9% after G-CSF administration at a Treg/ T effector cell ratio of 1:1). In vitro expansion of Tregs isolated after G-CSF application with anti-CD3 and anti-CD28 dynabeads in the presence of interleukin-2 led to comparable cell numbers as for the stimulation of control Treg cells. However, differences could be detected for the thymic derived marker molecule CD31 and those associated with activation (LAP, CD69, CD62L) and migration (CxCR3) as detected by FACS analysis. Our results show no significant differences regarding suppressive function and stability of Tregs isolated from stem cell donors before and after G-CSF administration. These data support the application of G-CSF mobilized Tregs for clinical trials which in turn opens up new possibilities for the adoptive transfer of Tregs in HSCT.

Disclosures:
No relevant conflicts of interest to declare.
Effects of Prophylactic Defibrotide for Veno-Occlusive Disease in Hematopoietic Stem Cell Transplantation
Authors: Park Meerim , Park Hyeon Jin , Eom Hyeon-Seok , Park Jeong A , Lim Yeon Jung , Yoon Jong Hyung , Lee Hye Won , Yun Tak , Park Byung-Kiu 
Published: October 2019

Abstract
Abstract 4517

Background:
Defibrotide has recently been successfully used to treat veno-occlusive disease (VOD). We report the use of intravenous defibrotide for VOD prophylaxis in 49 patients who received hematopoietic stem cell transplantation (HSCT). This group was compared with a control group of 49 patients who underwent HSCT without defibrotide prophylaxis.

Patients and methods:
Data were collected retrospectively on 98 patients undergoing HSCT at the National Cancer Center, Goyang, Korea, between August 2005 and July 2008. During this period, defibrotide was randomly supplied to 49 patients free of charge by the compassionate use program. For 49 patients, VOD prophylaxis with 200 to 400 mg of defibrotide (25 mg/kg/day in children weighing less than 30 kg) administered intravenously 3 or 4 times daily was initiated from the start of conditioning and continued until 28 days post-HSCT.

Results:
VOD was diagnosed in 5 of 98 total patients (5.1%) at a median of 11 days post-HSCT (range: 10–14 days). VOD was graded as mild in 2 patients, moderate in 1, and severe in 2. The incidences of engraftment syndrome, thrombotic microangiopathy and VOD were not significantly different between the defibrotide group and the control group, although absolute incidences of each syndrome were lower in the defibrotide group. Patients who received prophylactic defibrotide showed earlier platelet engraftment (p<0.01) and lower activated prothrombin time (p=0.02) after HSCT. There was no day 100 treatment-related mortality (TRM) in the defibrotide group. Day 100 TRM occurred in two patients who did not receive defibrotide for VOD prophylaxis during allogeneic HSCT. Each patient was diagnosed with mild or severe VOD, and the direct causes of death were infection or VOD. Of the patients who were diagnosed with VOD, the overall survival at 100 days post HSCT was significantly higher in the defibrotide group compared to the control group (100% vs. 33.3 ± 27.2%, p<0.01). Defibrotide was well tolerated and was not discontinued in any patients. There was no grade 3 or 4 toxicity related to defibrotide or any worsening of clinical bleeding.

Conclusions:
Our data showed that the use of prophylactic defibrotide is safe and may have beneficial effects of defibrotide on hepatic endothelial damage following HSCT, with no TRM in defibrotide group. Considering there is no consensus on when to start treatment if a patient shows clinical features suggesting VOD, it could be important to use prophylactic defibrotide in advance for improved VOD outcomes.

Disclosures:
No relevant conflicts of interest to declare.
Autologous Stem Cell Transplantation for NHL: 16 Years Follow- up
Authors: Guillermo Cecilia , Lipschutz Judith , Zunino Juan , Topolansky Laura , Rocca Alejandra , Stevenazzi Mariana , Diaz Andrea , Perdomo Susana , Perdomo Ana , Lavagna Graciela M , Isaurralde Hugo , Diaz Lilian , Nese Martha 
Published: October 2019

Abstract
Abstract 4520
Between June 1995 and June 2011, we performed 133 Autologous Stem Cell Transplantation (ASCT) in patients with NHL. Long term results of treatment and outcome were analyzed. The data presented were obtained from the Statistical Center of the Center for International Blood and Marrow Transplant Research. Median age was 48 years (range 18–65), 9% ≥ 60 years, 77 males and 55 females. The histology was: 98 (74%) B NHL, Follicular Lymphoma (FL) 26 (20%), Diffuse large B Cell Lymphoma (DLBCL) 47 (35%), Primary Mediastinal Large B Cell Lymphoma 12 (9%), Mantle cell Lymphoma (MCL) 6(4.5%), Marginal Zone Lymphoma 4 (3%), Small Lymphoplasmocytic 2 (1.5%), precursor B Lymphoplasmoblastic 1 (1%), and 35 (26%) T NHL, Peripheral T Cell Lymphomas NOS 12 (9 %), Anaplastic LC, T/N Cell systemic 9 (6.5%), Anaplastic LC, T/N Cell cutaneous 1(1%) and T/NK Cell Lymphoma 7 (5%), Angioinmunoblastic T Cell Lymphoma 2 (1.5%), Mycosis Fungoides 2 (1.5%), other T Lymphoma 2 (1.5%). We emphasize that 59 % of the patients had advanced disease at the moment of the transplant: 41% CR1, 11% CR2, 5% REL1, 8% primary induction failure, other 33% (CR3+, PR, and REL2+), and 2% missing/unknown. Condition regimens were BEAC 88%, BEAM 10% and CBV 2%. Stem cells source: 74% peripheral blood stem cells (PBSC) and 26% received bone marrow (BM) and PBSC.
Overall survival was 86% (95 CI, 80–92) at 1 year, 78% (95 CI, 70–85) at 3 years and 71% (95 CI, 62–79) at 5 years. Median follow up of survivors was 96 months (3–181). Primary cause of death was relapsed or progression.
We concluded that high dose therapy and Autologous Stem Cell Transplantation is an effective and feasible therapy for patients with high risk NHL. However relapse continues to be the most common cause of treatment failure. Newer strategies such monoclonal antibodody, require further investigation but early reports appear promising in reducing relapses in ASCT.

Disclosures:
No relevant conflicts of interest to declare.
Kinetics of Engraftment and Predictors of Outcome in Patients with Advanced Chronic Lymphocytic Leukaemia (CLL) After Treatment with Low Dose Total Body Irradiation (TBI) Followed by related or unrelated hematopoietic Stem Cell Transplantation
Authors: Hebenstreit Karin , Iacobelli Simona , Eisfeld Ann-Kathrin , Pfrepper Christian , Lange Thoralf , Krahl Rainer , Al-Ali Haifa K. , Becker Cornelia , Pönisch Wolfram , Behre Gerhard , Niederwieser Dietger 
Published: October 2019

Abstract
Abstract 4532

Introduction:
In an unicenter analysis we investigated the impact of allogeneic hematopoietic cell transplantation (RIC-HCT) after reduced intensity conditioning on the kinetics of engraftment and predictor of outcome in 50 patients with advanced CLL.

Patient and Methods:
Patients in advanced stage CLL (n=50) received Fludarabine 30mg/m2 on day -4 to -2 and 2Gy TBI on day 0 followed by cyclosporine and MMF from unrelated (n=40) or related (n=10) donors between June 1999 and March 2010 at the University of Leipzig. The majority of patients (n= 35) were male, had Binet C (n=37, 74%) at RIC-HCT. The median age was 58 (range 44–69) years. Of 48 patients for whom cytogenetic analyses were available, 22 (46%) had unfavourable cytogenetics including del 17 or del 11q. Three (6%) patients had CR, 27 (54%) PR, 7 (14%) progressive disease, 12 (24%) stable disease and one (5%) relapse at RIC-HCT. Resistance to first line therapy was present in 25 (50%) patients, whereas 12 (24%) were resistant to Fludarabine. Richter’s transformation was found in six patients (12%). Chimerism was detected in bone marrow and peripheral blood on T-lymphocytes and B-lymphocyte subpopulation after sorting at monthly intervals in the posttransplant period and than in 6 months interval.

Results:
Hematological toxicities after RIC-HCT were moderate. The majority of patients (96%) engrafted with neutrophiles &gt;500/μ L median at day 22 after HCT. Six (12%) and 15 (30%) patients maintained absolute neutrophil counts (ANC) &gt;0,5 × 109/L and platelet counts &gt;50 × 109/L, respectively. T-cell donor chimerism increased to &gt;95% at day 56 and B-cell donor chimerism to 94% at day +360, respectively. B-CLL cells disappeared completely on day +360 (median 0%). Overall survival (OS) at 4 years was 51%, Non relapse related mortality (NRM) 30%, Progression-free survival 33% and progression/relapse 37%. The most common causes of NRM were GvHD (n= 7; 14%) and sepsis (n=3, 6%). Factors significantly associated with increased risk of relapse/progression were intermediate/advanced disease vs. CR/PR1 (p=0.022) and lymphocytes ≥ 5 × 109/L vs. &lt; 5 × 109/L (18% vs. 58%, p= 0.00) at 12 months in univariate analysis.

Conclusion:
Full donor T-cell chimerism was reached early after HCT, while B-cell reconstitution was observed only 1.5 years after RIC-HCT despite the absence of evident disease by 360 days after RIC-HCT. Best predictor for Progression-free survival (PFS) was CR or PR1.

Disclosures:
Pönisch: Mundipharma: Honoraria, Research Funding.
Increased Production of the Inflammatory Cytokine IL12p70 by Activated Human Blood Dendritic Cells Is Associated with Increased Severity of Acute Graft Vs Host Disease Post Allogeneic Hemopoietic Cell Transplantation
Authors: Sartor Mary M , Lau Jenny , Gottlieb David , Bradstock Kenneth F 
Published: October 2019

Abstract
Abstract 4539

Introduction:
Dendritic cells (DC) are centrally involved in the initiation of acute graft versus host disease (GVHD) after allogeneic hemopoietic cell transplantation (alloHCT). We have shown that the activation status of peripheral blood CD11c+ myeloid DC, as assessed by CMRF-44 antigen expression, is highly associated with the occurrence and severity of acute GvHD (Transplantation 2007;83: 839–846). However, very little is known about the function of DC after human alloHCT. We examined the relationship between DC functional properties and the severity of acute GvHD.

Methods:
Peripheral blood CD11c+ myeloid DC from 12 patients were studied weekly up to 8 weeks post transplant for production of IL2, IL4, INFg, IL10 and IL12 using an intracellular cytokine flow assay. Mixed lymphocyte reactions using flow sorted patient DC and third party T cells were used to assess allogeneic immune responses induced by recipient DC in 5 patients.

Results:
IL12 was the only cytokine detected in post transplant DC. Five of 12 patients developed grade II-IV GvHD, the remaining 7 patients developed either no aGvHD or only grade I. In comparison with pre-transplant levels of expression of IL12, patients with grade II-IV GVHD had a significantly higher percentage of CD11c+ DC expressing IL12 (median 15.1%, range 11.2–20.9%) as compared to patients with grade 0-I GvHD in whom there was no change from baseline values (median 6.6% range 2.8–8.9%) p=0.0025. Increased expression of IL12 was observed in CD11c+ DC commencing at day 25 post transplant. Interestingly, analysis of 5 paired samples comparing sorted DC from normal donors with sorted DC from peripheral blood at day +30 post-transplant showed a marked reduction (measured by thymidine uptake from 60% to 90%) in the capacity of post-transplant donor DCs to stimulate 3rd party lymphocyte proliferation. None of these patients developed clinically significant aGvHD.

Conclusion:
Production of IL-12p70 by CD11c+ myeloid DC correlates with severity of GvHD despite impaired capacity of these cells to elicit third party proliferative responses.

Disclosures:
No relevant conflicts of interest to declare.