bluuebunny/crossref_metadata_2025 huggingface dataset:
Procoagulant Activity of Laminaria Japonica Derived Fucoidan (BAX513) Depends on the Interaction with the C-Terminus of Tissue Factor Pathway Inhibitor
Authors: Palige Michael , Redl Christoph , Knappe Sabine , Ehrlich Hartmut J. , Dockal Michael , Scheiflinger Friedrich
Published: October 2019

Abstract 4417

BAX513, a fucoidan derived from the brown seaweed Laminaria japonica, and other non-anticoagulant sulfated polysaccharides (NASPs) improve coagulation in hemophilic blood and plasma. Fucoidans are heterogeneous, polysulfated molecules with procoagulant activities in a wide concentration range. Tissue factor pathway inhibitor (TFPI) has been described as a potential target for the procoagulant activity of NASPs (Liu et al. Thromb Haemost 2006; 95:68). In the current study, we investigated the interaction of BAX513 with TFPI proteins to gain a detailed understanding of the mechanism of action of BAX513. We used calibrated automated thrombography to monitor the activity of BAX513 in normal, FX and TFPI-deficient plasma. TFPI plasma levels were varied by the addition of truncated TFPI (TFPI1-160) and TFPI-domain specific antibodies. Initiating thrombin generation by addition of FXa to plasma deficient in both, FX and FVIII-showed a BAX513-dose dependent increase of thrombin generation, which was completely abolished when TFPI-specific polyclonal antibodies were present. Furthermore, when full-length TFPI was inhibited in plasma and instead supplemented with increasing amounts of TFPI 1–160, BAX513 did not show any activity. The data are further supported by surface plasmon resonance experiments (BiaCore) exploring the BAX513-TFPI interaction. A high affinity interaction was only observed for BAX513 with full-length TFPI but not for BAX513 with TFPI1-160. Our findings support a mechanism of action in which BAX513 acts as a potent dose-dependent TFPI antagonist that requires the highly charged C-terminus of TFPI to unfold its full potential. Understanding the mechanism of action of BAX513 supports the development of BAX513 as a promising new therapeutic for hemophiliacs and FVIII-inhibitor patients.

Disclosures:

Palige: Baxter Innovations GmbH: Employment. Redl:Baxter Innovations GmbH: Employment. Knappe:Baxter Innovations GmbH: Employment. Ehrlich:Baxter Innovations GmbH: Employment. Dockal:Baxter Innovations GmbH: Employment. Scheiflinger:Baxter Innovations GmbH: Employment.
All-Trans Retinoic Acid Modulates the Balance of ADAMTS13 and VWF In Human Microvascular Endothelial Cells
Authors: Wang Anyou , Yu Ziqiang , Dong Ningzheng , Ma Zhenni , Zhang Jingyu , Ruan Changgeng
Published: October 2019

Abstract 4424

Introduction:

All-trans retinoic acid (ATRA) can modulate the hypercoagulable state of human microvascular endothelial cell, which has been confirmed by the reports that ATRA can chang the expression of some procoagulant or anticoagulant factor, such as upregulation tissue factor and downregulation thrombomodulin. While, the balance between ADAMTS13 and von Willebrand factor (VWF) of microvascular endothelial cell, very related with the formation of microvascular platelet thrombsis, play an important role on the controlling the hypercoagulable state of microvascular endothelial cell. And it has been reported that the balance interrupted by some inflammatory cytokines may contribute to microvascular platelet thrombsis. In this study, to better understand the antithrombotic property of ATRA, we investigated whether ATRA may affect the balance.

Materials and Method:

We observed the effects of ATRA (compared to TNF-α) on the expression of ADAMTS13mRNA in human microvascular endothelial cell (HMEC-1 cell line) by real-time polymerase chain reaction amplification (RT-PCR). The levels of ADAMTS13 and VWF antigen in the conditioned medium of HMEC-1 cells were also detected by western blot or enzyme-linked immunosorbent assay (ELISA), and the proteolytic activity of ADAMTS13 was also determined by using GST-VWF73-His peptide as a specific substrate.

Result:

In a dose-dependent manner, ATRA significantly upregulated the expression of ADAMTS13mRNA which could be almost up to 4.63 times with 10uM ATRA, while TNF-α inhibited ADAMTS13mRNA expression which could drop to 41% of the control group with 10ng/ml TNF-α. ATRA also reverse the inhibition of TNF-α to ADAMTS13mRNA expression. The results were confirmed from the levels of ADAMTS13 protein and its activity, while ATRA had no significant affection on triggering release of VWF known as a specific substrate of ADAMTS13.

Conclusion:

This study provides the evidence that ATRA modulates the balance of ADAMTS13 and VWF in human microvascular endothelial cell, which may be a very relevant compartment for the antithrombotic property of ATRA.

Disclosures:

No relevant conflicts of interest to declare.
Preliminary Study of the Relationship Between EPO Receptor and Autoantibody on the Membrane of Erythropoietic Cells of the Patients with BMMNC- Coomb's Test(+) Hemocytopenia
Authors: Fu Rong , Liu Hui , Shao Zonghong , Li Lijuan
Published: October 2019

Abstract 4436

Objective

To observe the relationship between EPO receptor (EPOR) and autoantibody(IgG) on the membrane of erythropoietic cells of the patients with bone marrow mononuclear cell Coomb's(BMMNC-Coomb's) test(+) immuno-related pancytopenia(IRP),and then explore the probable autoantigens of autoantibodies in IRP.

Methods

42 newly diagnosed IRP patients (14 with IgG autoantibody on erythropoietic cells and 28 without this autoantibody) and 11 healthy donors as controls were enrolled in this study. EPOR on their nuclear erythrocyte were tested with flow cytometry. Their clinical data were also analyzed.

Results

(1) The level of EPOR of IgG(+) arm(1.59±0.87)% was significantly lower than that of IgG(-) arm (4.58±4.09)%(P<0.01), and the level of EPOR of IgG(-) arm was significantly higher than that of normal controls(2.27±1.76)%(P<0.05) G The level of EPOR of IRP patients was negatively correlated with their IgG autoantibody(r=-0.543,P=0.000) and its regression equation was Y(EPOR)=0.040-0.335X(IgG); (2) Reticulocyte percentage of IgG(+) arm (1.55±0.64)% was significantly lower than that of IgG(-) arm (2.41±1.42)%(P<0.05). The level of EPOR of IRP patients was positively correlated with reticulocyte percentage(r=0.346,P=0.029); (3)The serum level of indirect bilirubin(IDBIL) of IgG(+) arm was normal. While there were six patients (21.4%) whose serum level of IDBIL were higher than those of normal controls in IgG(-) arm.

Conclusion

The autoantibody(IgG) of some IRP patients might block or competitively inhibit the EPOR on the membrane of erythropoietic cells. It could inhibit the proliferation and differentiation of erythropoiesis in early phase.

Disclosures:

No relevant conflicts of interest to declare.
Dasatinib Compared with Imatinib In Newly Diagnosed Chronic Myelogenous Leukemia In Chronic Phase (CML-CP):Results of Japanese Subset Analysis In DASISION Trial.
Authors: Ogura Michinori , Nakamae Hirohisa , Fujisawa Shin , Ishizawa Kenichi , Taniwaki Masafumi , Utsunomiya Atae , Matsue Kosei , Tamura Kazuo , Usuki Kensuke , Tanimoto Mitsune , Ishida Yoji , Akiyama Hideki , Sakai Rika , Kasai Masanobu , Hino Masayuki , Ueda Kyoji , Seriu Taku , Bradley-Garelik M. Brigid , Zhu Chao
Published: October 2019

Abstract 4484

Background:

Dasatinib is a highly potent BCR-ABL kinase inhibitor. The previous report from the global DASISION trial showed dasatinib 100 mg once daily resulted in significantly higher and faster rates of complete cytogenetic response (CCyR) and major molecular response (MMR) compared with imatinib; both treatment arms were well-tolerated (N Engl J Med. 2010;362:2260-70). The objective of this subset analysis was to assess the efficacy and safety of dasatinib compared with imatinib in the Japanese population.

Methods:

Forty-nine Japanese patients (total 519 pts) with newly diagnosed CML-CP were randomly assigned to receive dasatinib 100 mg QD or imatinib 400 mg QD. Confirmed CCyR (cCCyR; CCyR on 2 consecutive assessments at least 28 days apart) was the primary efficacy endpoint with MMR as an important secondary endpoint. The safety profiles were also evaluated.

Results:

Minimum follow-up time and median treatment duration were 12 months and 15 months, respectively. Twenty-six patients with median age 56 (range, 21–70) years were treated with dasatinib and 23 patients with median age 52 (range, 22–77) years were treated with imatinib. Overall 89% of patients receiving dasatinib and 83% of patients receiving imatinib continue to receive treatment. The cCCyR rate by 12 months (primary endpoint), CCyR rate by 12 months and MMR rate at any time in dasatinib arm were higher than those in imatinib for Japanese patients (96% vs 70%, 96% vs 78%, and 73% vs 48%, respectively). Grade 3/4 cytopenias in dasatinib arm and imatinib arm were as follows: anemia (8% vs 4%), neutropenia (27% vs 39%), and thrombocytopenia (8% vs 9%). Non-hematologic and drug-related adverse events occurring in ≥10% of patients are shown as Table. No deaths were reported in either group. Drug-related serious adverse events were rarely reported and all events were not severe (Grade 1–2, including vomiting, hypoxia and cardiomyopathy in dasatinib arm).

Conclusion:

Dasatinib showed higher rates of cCCyR and MMR compared with imatinib. Both treatments were well tolerated. Given the predictive value of 12 months cCCyR, dasatinib may improve long-term outcomes in Japanese patients with newly diagnosed CML-CP.

Disclosures:

Ueda: Bristol-Myers K.K.: Employment. Seriu:Bristol-Myers K.K.: Employment. Bradley-Garelik:Bristol-Myers Squibb: Employment. Zhu:Bristol-Myers Squibb: Employment.
Pregnancy out Come In the Perspective of Chronic Myeloid Leukemia Patient on Imatinib Therapy: An Experience From India.
Authors: Mukhopadhyay Soma , Mukhopadhyay Ashis , Dasgupta Swati , Dey Subhasish , Majumdar Deboshree , Sarkar Sanjoy , Gharami Firoj , Konar Sukanta , Ray Ujjal
Published: October 2019

Abstract 4493

Objective:

Now that imatinib is being used to treat thousands of chronic myeloid leukemia (CML) patients for more than 10 year it is highly probable that many patients will get pregnant during its use. Company warns against any such use. But the fact remains that there is need for planned pregnancies in indicated cases. So we selected few cases both male and female for such pregnancies by interrupting treatment and following the pregnancy closely. Their outcome was studied so that we have an idea about what best could be suggested in such instance.

Patients and Methods:

From November 2002 to May 2010, 634 patients with CML in any stage of the disease were treated with imatinib at our tertiary cancer research institute. Among them 274 were female and 221 were of 17 to 50 years age group. We report information of 8 accidental pregnancies and 10 planned pregnancies involving 18 patients (10 females and 8 males) where pregnancy had occurred while receiving imatinib for the treatment of CML.

Observations:

Among 10 pregnancies reported in female sufferers there were two spontaneous and one elective abortion all in unplanned group and seven live births including one twin. There was one case of hypospadius which could be surgically corrected. Among the eight male patients whose wives conceived four pregnancies were planned, there was one spontaneous abortion, two elective abortions. The conceptions resulted in the birth of five healthy babies (two females). There was no other identifiable congenital anomaly.

Conclusion:

In the patients, who do become pregnant while on treatment, balancing the risk to the fetus of continuing imatinib versus the risk to the mother of interrupting treatment remains difficult. From the fetal perspective, imatinib should be discontinued due to the potential risk of serious developmental abnormalities; from the maternal perspective also there is chance of drug interruption induced cytogenetic relapse. CML patients, in the child bearing age group can do bear children. There was no major foetal anomalies which was documented. In the case of female patients the drug was withheld during the first trimester for concerns regarding potential teratogenecity. In both male and female, in the case of a planned pregnancy the drug should preferably be stopped one month before the time of expected conception.

Disclosures:

No relevant conflicts of interest to declare.
Relapse of Lymphoma After Allogeneic Hematopoietic Cell Transplantation: Management Strategies and Outcome.
Authors: Wudhikarn Kitsada , Cao Qing , Brunstein Claudio , Bachanova Veronika , Burns Linda J , Weisdorf Daniel J.
Published: October 2019

Abstract 4503

Background&Objective:

The management of lymphoma which has relapsed after allogeneic hematopoietic cell transplantation (HCT) is difficult. Therapeutic options may include donor lymphocyte infusion (DLI), withdrawal of immunosuppression, rituximab, chemotherapy, radiation, immunotherapy and experimental treatments, but response and survival are uncertain.

Result:

We analyzed 72 patients with relapsed or progressive lymphoma after allogeneic HCT (1991-2007); 44 non-Hodgkin's lymphoma (7 mantle cell, 5 indolent, 19 diffuse large B [DLBCL], and 13 T/NK cell) and 28 Hodgkin's lymphoma (HL). At HCT, 62 patients (86%) were in remission (22 CR, 40 PR); nine (13%) had progressive disease and one had stable disease. Conditioning was myeloablative (n=17) or reduced intensity conditioning (RIC) (n=55). Relapse or progression occurred at a median of 99 days (0-1898 days; 25–75% 43–194 days). At relapse, 41 (57%) had extensive nodal disease and 56 (78%) had extranodal organ involvement. Management of relapse included: no therapy (n=5, 7%), reduction of immunosuppression (RIS) (n=58, 81%), chemotherapy alone (n=14, 19%), immunotherapy alone (n=20, 28%), combined chemo-immunotherapy (n=7, 10%) donor lymphocyte infusion (DLI) (n=7, 10%), second allogeneic HCT (n=2, 3%), radiation (n=23, 32%) and other therapy (n=7, 8%). Twenty-four patients achieved a remission (15 CR, 9 PR) to the initial salvage therapy: RIS (n=13), combined chemo-immunotherapy(n=5), chemotherapy alone (n=2), radiation (n=3) or DLI (n=1). Overall, 40 (56%) patients responded (30 CR, 8 PR, 2 stable disease) with a median post-relapse overall survival of 28 months (range 0–147 months). At 3 years following relapse, 44% (95% CI, 32–56%) survive. Favorable prognostic factors for improved 3-year survival after relapse include stage I-III at relapse, single site of relapse and most importantly, later relapse (>100 days after HCT [3 year survival 53% vs. 36%, p=0.02]).

Conclusion:

The overall prognosis of relapsed lymphoma after allogeneic HCT differs based upon extent of disease at relapse and time of relapse. Appropriate therapeutic approaches (RIS, combined chemo-immunotherapy, radiation or DLI) in these high risk, post transplant relapsed patients can yield promising outcome. Prospective study of post-relapse therapy to determine the most effective management strategies are warranted.

Disclosures:

No relevant conflicts of interest to declare.
CMV Specific T Cells Prevent Progression From Low to High Level Viremia In D+R+ but Not D-R+ Patients.
Authors: Hoegh-Petersen Mette , Roa Lina , Liu Yiping , Zhou Feng , Ugarte-Torres Alejandra , Louie Polly , Fonseca Kevin , Khan Faisal , Storek Jan
Published: October 2019

Abstract 4532

Background.

Cytomegalovirus (CMV) reactivates (becomes detectable in blood) in most seropositive hematopoietic cell transplant (HCT) recipients. In some reactivating patients, low level viremia (<50,000 DNA copies/ml plasma, ie, less than our institutional threshold for preemtive therapy) progresses to high level viremia or CMV disease, which is potentially fatal. We hypothesized that low level viremia progresses in patients with low specific T cell counts and spontaneously resolves in patients with high specific T cell counts.

Methods.

In 30 CMV seropositive HCT recipients monitored weekly for reactivation by real-time PCR, blood was drawn for specific T cell counts within 4 days from the first episode of low level viremia. Fourteen patients received grafts from seropositive donors (D+R+), and 16 patients from seronegative donors (D-R+). Mononuclear cells were stimulated overnight with CMV lysate, pp65 overlapping peptides, no stimulus (negative control) or staphylococcal enterotoxin B (positive control). T cells producing IFNγ, TNFα and/or IL2 were enumerated by flow cytometry.

Results.

Among D+R+ patients, counts of CMV lysate and pp65 specific CD4 T cells producing IFNγ and TNFα (and not IL2) were higher in patients with spontaneous resolution than patients with progression (p=0.02 for CMV lysate, p=0.004 for pp65). Also, there was an inverse correlation between pp65 specific CD8 T cells producing IFNγ and TNFα and peak viremia (r=-0.94, p=0.005) in D+R+ patients who progressed to high level viremia/disease. In contrast, among D-R+ patients, CMV lysate and pp65 specific T cell counts were similar in patients with spontaneous resolution and patients with progression, and there was no correlation between specific T cell counts and peak viremia.

Conclusion.

CMV specific T cells play a role in preventing progression from low to high level CMV reactivation/disease in D+R+ patients. Other immune mechanisms (eg, NK cells?) play the role in D-R+ patients.

Disclosures:

No relevant conflicts of interest to declare.
Transient but Reversible Activation Defect of T-Cell Responses Against Cytomegalovirus and Adenovirus through Immunosuppresion with Calcineurin Inhibitors.
Authors: Neinhaus Cornelia , Opherk Kathrin , Kayser Simone , Leibold Joseph , Feucht Judith , Lang Peter , Handgretinger Rupert , Feuchtinger Tobias F.
Published: October 2019

Abstract 4542

Immunosuppressive treatment is widely used, especially after allogeneic stem cell transplantation (HSCT) to prevent or treat graft versus host disease (GvHD). Common drugs are Ciclosporin A or Tacrolimus in combination with steroids. However, immunosuppressive treatment and the underlying conditions are associated with an increased risk of viral reactivations with persistent pathogens like cytomegalovirus or adenovirus. In the absence of a protective immune response, virus infection remains a life-threatening complication after HSCT. Here we investigated the antiviral T-cell response (n=12) after ex vivo exposition with Ciclosporin A, Tacrolimus, Prednisolone or Mycophenolate at time of immunosuppression, 24 hours and 72 hours later. Analysis has been done with IFNgamma Elispot assays, confirmed by intracellular cytokine staining in flow cytometry and analysis of T-cell proliferation detected by CFSE. The antiviral T-cell response is suppressed after 24 hours using normal serum concentrations (100-200ng/ml) of cyclosporine A. T-cell annergy, induced by cyclosporine, could be reversible, after 72 without immunosuppression. Tacrolimus has a stronger immunosuppressive effect on T-cell activation within the same time and even low levels of 1ng/ml induce T-cell suppression after 72 hours. Peak levels of calcineurin inhibitors even suppressed the T-cell response to superantigens like staphylococcal antigen B or PHA. As expected, Prednisolone had a short and dose dependend effect on T-cell activation. Mycophenolate has a mild effect on the activation of virus-specific T-cells. However, all three drugs induced a significant reduction in Ag-specific T-cell proliferation. In conclusion, Interferonγ detection in virus-specific T-cells is a good diagnostic tool for clinicians to monitor the risk of viral complications in immunosuppressed patients. Tacrolimus, Cyclosporin A, Prednisolone and Mycophenolate induce an activation defect in Ag-specific T-cells with decreasing severity. The effect is reversible and corresponds to high or low serum levels.

Disclosures:

No relevant conflicts of interest to declare.
Thoax CT Scan as Invasive Fungal Infection (IFI) Screening In Neutropenic Patients, a Prospective-Retrospective Experience of It Impact on Therapeutic Intervention.
Authors: Sevilla Sebastian , Kusminsky Gustavo Daniel , Damiano Mario Atilio , Rizzo Miguel , Trucco Jose
Published: October 2019

Abstract 4554

Introduction:

Persistent fever in high risk neutropenic patients (HRNF) after day 5 of empiric treatment is a sign of high susceptibility for IFI with elevated morbidity and mortality. Diagnostic tools in this setting are inaccurate to determine the occurrence of IFI and most patients start with empiric antifungal agents. Drugs are usually associated with increasing costs and toxicity. It is challenging to establish the population of patients in whom in spite of persistent fever and neutropenia, avoidance of antifungal treatment is a reasonable strategy.

Methods:

We have prospectively allocated 229 HRNF patients in different empiric antimicrobial regimens over a 4.5 year period. In a retrospective revision, there were 33 patients with persistent fever on day 5 of empirical antimicrobial treatment and no evident new infection episode or clinical impairment. In 28 patients, a thorax CT scan was performed as part of the evaluation of persistent fever. The clinical outcome was evaluated regarding the presence or absence of pulmonary infiltrates in the CT scans. Initial empiric antifungal treatment, transfusions, days in hospital, days with neutropenia, antimicrobial treatment, and days with fever were evaluated.

Results:

Nineteen patients (68%) of 28 presented with pulmonary infiltrates. All of them received antifungal treatment. In 9 patients with normal CT scan antifungal treatment was deferred. The difference of the decision in not giving antifungals according CT scans was highly significant (p <0,0001). Transfusions of red blood cells and platelets were significantly less in the group of normal scans (p 0,0004 and 0,005 respectively). Antimicrobial treatment, days in hospital and days with fever were not significantly different in both groups. There was one death in the normal scan group due to relapse. Mortality was not significantly different in both groups.

Conclusion:

In HRNP, normal thorax CT scan changed the clinical decision in not starting antifungal treatment in spite of persistent fever. There was no difference in mortality with patients under antifungal treatment, allowing continuing with this strategy in more patients in the future.

Disclosures:

No relevant conflicts of interest to declare.
Reduced Intensity Conditioning Allogeneic Hematopoietic Stem Cell Transplantation for Low Grade Non Hodgkin's Lymphoma: Preliminary Results at Hospital De Clínicas De Porto Alegre (HCPA), Porto Alegre, RS, Brazil.
Authors: Araujo Beatriz Stela Gomes de Souza Pitombeira , Paz Alessandra Aparecida , Ferreira Clarissa Barros , Boffo Manoela Santos , Ottoni Erica Lammerhirt , Fischer Gustavo Brandão , Daudt Liane Esteves , Silla Lucia
Published: October 2019

Abstract 4557

INTRODUCTION:

Advanced low grade non-Hodgkin Lymphomas have many and varied treatment options. For patients with relapsed or refractory disease, outcome following only chemotherapy protocols remains poor. The standard Conditioning Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) might be an alternative for relapsed low grade non-Hodgkin lymphomas, but its high Transplant Related Mortality (TRM), varying from 24% to 54%, accounts for a lower overall survival in this group. Reduced Intensity Conditioning (RIC) HSCT has been shown to be a good alternative to myeloablative regimens, with lower TRM, Graft versus Lymphoma Effect, and appears to be associated with improved outcome, presenting a 3 years disease free survival and overall survival from 32% to 66% and from 32% to 73%, respectively, depending on factors such as disease stage before transplantation.

PATIENTS, METHODS AND RESULTS:

Sixteen heavily pretreated patients with low grade non-Hodgkin Lymphoma (NHL) were submitted to RIC HSCT with matched related donors at the Bone Marrow Transplantation Unit at HCPA between the period of September 2002 to October 2009. The mean age was 43 years and were 7 patients male and 9 female. Fifty percent (8) of the patients had refractory disease pre-HSCT, 43.8% (7) were at third complete remission (CR) and 6.2% (1) at second CR. Median of CD34/kg infused was 10.7 × 106 and the median of days for engraftment was 14.53. Conditioning regimens were Fludarabine + Cyclophosphamide in 3 patients, Fludarabine + Melphalan in 7, Fludarabine + Cyclophosphamide + Rituximab in 2, Fludarabine + Cyclophosphamide + Melphalan in 1, Cyclophosphamide + Total Body Irradiation (TBI) in 1 and Fludarabine + Cyclophosphamide + TBI in 2. Acute Graft versus Host Disease (aGVHD) occurred in 50%, with 37.5% of those were steroids resistant; and 62.5% of the patients evolved to chronic GVHD, with 80% of them classified as extensive. Chimerism from 9 patients was analyzed, and 89% of them were complete. Mean overall survival was 2.95 years; 62.5% of patients were alive within 1 year and 40% within 3 years. TRM was 6.66%. Eight patients died, five of them were transplanted with refractory disease. Only 2 patients died of relapse, 5 of infections and 1 of aGVHD.

CONCLUSION:

Low grade NHL patients treated at HCPA with RIC HSCT had an overall survival and TRM comparable to the observed at other centers, appearing as a good alternative for this heavily treated group of patients with less toxicity than myeloablative regimens.

Disclosures:

No relevant conflicts of interest to declare.
The Effect of KIR B/X Genotype and Missing KIR Ligands In Haploidentical Hematopoietic Stem Cell Transplantation.
Authors: Sun Aining , He Jun , Zhou Lina , Depei Wu , Li Weiyang , Bao Xiaojing
Published: October 2019

Abstract 4578

Objective:

To study the prognostic impact of missing ligands for inhibitory killer-immunoglobulin receptor(KIR) and donor KIR genotype in haploidentical hematopoietic stem cell transplantation(HSCT).

Methods:

HLA genotype of 14 patients (ALL 8 cases, AML 3 cases, CML 1 cases, NHL 1 cases and granulocytic sarcoma 1 case) and their donors was determined by polymerase chain reaction sequence oligonucleotide probes (PCR-SSOP) and sequence specific primers (PCR-SSP). The KIR genotype was determined by PCR-SSP.

Results:

1. Donors were divided into those with (n=9) KIR B/X and those with (n=5) KIR A/A. We found that the 2-year OS rate was higher with KIR B/X versus KIR A/A donors (33.3% vs 20%, P=0.308). Of the B/X group, most donors (77.8%) expressed activated KIR (aKIR) with 2DS4 or 2DS1. The 2-year OS rate was higher for those (n=5) with 1–2 aKIR than those (n=4) with 3–4 aKIR group (40% vs 25%). The GVHD incidence was slightly lower with 1–2 aKIR versus 3–4 aKIR donors (40% vs 50%).

2. Patients were divided into those with (n=11) and those without (n=3) missing 1 or more HLA class I ligands for donor inhibitory KIR. The 2-year OS rate for patients with and without missing KIR ligands was 36.4% and 0%,respectively(P=0.680)

Conclusion:

In haploidentical HSCT, donor and recipient should be performed the high resolution HLA-Cw genotyping and KIR genotyping (A/A and B/x). Using donors with KIR B/X genotype or missing KIR ligands may be associated with improved OS and decreased GVHD.

Disclosures:

No relevant conflicts of interest to declare.
The Use of Double-Unit Umbilical Cord Blood Transplantation In Adults with Hematologic Disease.
Authors: Ma Xiao , Depei Wu , Sun Aining
Published: October 2019

Abstract 4579

Objective:

In this study, we explored the efficiency and toxicity of 38 cases of double-unit umbilical cord blood transplantation (CBT) in adults with hematologic disease.

Methods:

The tolerance; transplant related complications; survival rate and disease free survival rate were observed and analyzed. A nonmyeloablative conditioning regimen included cyclophosphamide, fludarabine and 2Gy TBI. Cyclosporine combined mycophenolate mofetil and ATG were used to prevent graft versus host disease (GVHD).

Results:

All these 38 patients tolerated the therapy well while two patients had graft failure. Severe acute GVHD was presented in 6 patients. Chronic GVHD was occurred in 16 patients. Fatal infection complications were occurred in 5 patients (including CMV idiopathic pneumonia in 2 patients) and 4 patients relapsed after transplantation. Neutrophil engraftment obtained on day +17 and platelet reconstitution occurred on day +42 on median. In the follow-up duration of 17 months on median, the expected 2-year relapse mortality was 17.95%; non-relapse mortality was 25.90%; overall survival was 60.80%, and disease free survival was 52.11%. Compare with 39 cases of haplo-identical HSCT in our department, significant delays in engraftment and lower severe GVHD occurred after CBT. There was no apparent difference between the risk of relapse and DFS in both groups.

Conclusion:

The use of double-unit CBT after reduced intensive conditioning therapy in adults with hematologic disease is an effective and safe treatment. Fatal infection and relapse are the main reasons of failure.

Disclosures:

No relevant conflicts of interest to declare.
Bortezomib and High-Dose Melphalan as a Novel Conditioning Regimen for Patients with POEMS Syndrome Undergoing Autologous Peripheral Blood Stem Cell Transplantation.
Authors: Tang XiaoWen , Shi Xiaolan , Hu Xiaohui , Xue Shengli , Qiu Huiying , Han Yue , Yan Lingzhi , Zhou Huifen , Liu Yuejun , Gu Bin , Sun Aining , Depei Wu
Published: October 2019

Abstract 4599

Background:

POEMS syndrome is a multisystem disorder associated with plasma cell dyscrasias which characterized by polyneuropathy (P), organomegaly (O), endocrinopathy (E), serum M-protein (M), and skin changes (S). Recently, we successfully treated two POEMS syndrome patients by using bortezomib-based regimen(VDD) and followed by autologous peripheral blood stem cell transplantation (APBSCT) with bortezomib combined with high-dose melphalan(Mel) conditioning regimen.

Methods:

According to the latest Mayo Clinic criteria, two patients’ diagnosis of POEMS syndrome could be comfirmed. Both of them had no response to the classical treatments, but achieved near complete remission(CR) after 4 cycles and 1 cycle of VDD respectively(Bortezomib 1.3–1.6 mg/m2/w×4 weeks; 40 mg of Liposomal doxorubicin on the fourth day of the first week; and 10 mg of dexamethasone during the initial 4 days of first cycle. Each course was at 21 days interval). Then, autologous peripheral blood stem cell collection was performed sucessfully (2.16×106/kg and 3×106/kg respectively) after mobilization by cyclophosphamide (3g/m2/d for 1 day) with subcutaneous G-CSF. APBSCT were performed approximately 1 month after stem cell collection. Two patients were conditioned with Bortezomib 1.6mg/m2(d-13,d-6,d+1,d+7)+Mel 200mg/m2(d-3)and Bortezomib 1.3 mg/m2(d-8,d-1,d+6,d+13)+ Mel 200mg/m2(d-2) respectively.

Results:

No toxic death or serious adverse effects occurred during APBSCT. Neutrophil and platelet recoveried at +10d, +27d and +7d, +20d respectively. Only case 1 patient developed mild hypoxemia during neutrophil engraftment period, but was successfully treated with corticosteroid and antibiotic therapy. The case 2 presented mucositis, diarrhea and nausea during neutropenic peroid. A follow-up period were 12 and 6 months respectively. Both of them achieved continuous CR. There was a dramatic improvement in clinical symptoms and serum VEGF levels in all two patients post VDD treatment including organomegaly (splenomegaly), M-protein, pericardiac/pleural effusion,ascites and polyneuropathy.

Conclusion:

This is the first report of POEMS patients treated by APBSCT with bortezomib and high dose Melphalan conditioning regimen. Our results suggest that Bortezomib is a new effective and relative safe therapeutic option for POEMS syndrome not only in the conventional treatment but also in APBSCT procedure.

Disclosures:

No relevant conflicts of interest to declare.
Epigenetic Downregulation of Secreted Frizzled-Related Proteins In Philadelphia Positive and Philadelphia Negative Myeloproliferative Neoplasms
Authors: Bennemann Karla , Schubert Claudia , Wilop Stefan , Brummendorf Tim H. , Galm Oliver , Jost Edgar
Published: October 2019

Abstract 4647

Introduction:

Hypermethylation of CpG islands in promoter regions is associated with transcriptional inactivation and represents an important mechanism of tumor suppressor gene silencing and plays a role in the pathogenesis of hematopoietic malignancies. Myeloproliferative neoplasms (MPN) constitute a heterogenous group of hematopoietic malignancies and comprise chronic myeloid leukemia (CML), polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF). BCR/ABL-negative MPN (Ph-MPN) are often characterized by an autoactivating mutation in the JAK2 tyrosine kinase (JAK2V617F) as well as additional mutations in the JAK-STAT pathway. Secreted Frizzled-related proteins (SFRPs) are antagonists of the Wnt signaling pathway. Consistent with the central role of the Wnt pathway for stem cell maintenance and differentiation of hematopoietic progenitors. Upregulation of Wnt-pathway has been described in haematopoietic malignancies and epigenetic downregulation of SFRPs has been shown in acute leukemia.

Methods:

We determined the promoter methylation status of SFRP1, -2, -4 and -5 in human cell lines and in 60 samples from MPN patients by methylation-specific PCR (MSP). The JAK2V617F mutation was assessed by allele-specific PCR.

Results:

Aberrant methylation among primary MPN samples was 5 % (3/60) for SFRP-1, 24 % (14/60) for SFRP-2, 2% (1/60) for SFRP-4 and 0% (0/60) for SFRP-5. The SFRP-2 promoter was methylated in 0% of CML (0/12), 27% of PV (3/11), 30% of ET (3/10) and 26% of PMF patient samples (7/27). There was no correlation between SFRP-2 hypermethylation and MPN subtype. We detected a trend for a correlation between methylation of the SFRP-2 promoter and presence of JAK2V617F mutation (p = 0,1).

Discussion:

Our data indicate that epigenetic dysregulation of the Wnt signaling pathway is a common event in Ph-MPN with aberrant methylation of at least one SFRP being detected in 30% of the primary patient samples with the most predominant member of the family, SFRP2 being methylated in about one third of the cases. Aberrant methylation of SFRPs may contribute to the pathogenesis of MPN by influencing stem cell maintenance and proliferation of hematopoietic progenitors.

Disclosures:

No relevant conflicts of interest to declare.
Anaphylactic Reaction After Replacement Therapy In Haemophilia B Patient with Inhibitor to Factor IX
Authors: Scaraggi Rosanna , Giordano Paola , Ettorre Cosimo Pietro , Malcangi Giuseppe , Marino Renato , Lassandro Giuseppe , De Mattia Domenico , Scaraggi Francesco Antonio
Published: October 2019

Abstract 4662

Replacement therapy is a very hard challenge in haemophilia B with inhibitor. We describe the case of a child with severe haemophilia B and with a family history positive for development of inhibitor to factor IX (FIX) and for occurrence of allergic reaction after replacement therapy. Genetic analysis demonstrated an almost complete deletion of FIX gene. The child received replacement therapy first in his life when he was 5 years old because of the occurrence of a large haematoma of the left thigh. He was treated with recombinant FIX concentrate at the dosage of 30 IU/kg daily for three days and no inhibitor to FIX was evidenced after this therapy. Two months later the patient was treated with a single dose of recombinant FIX concentrate at the same dosage for the occurrence of a post-traumatic joint bleeding of the right knee. An inhibitor towards FIX (1.7 B.U.) was detected two weeks after this treatment and confirmed in a subsequent analysis performed after ten days (1.4 B.U.). One month later the patient was hospitalized for a post traumatic hemarthrosis of the right shoulder. In this occasion it was planned to treat the patient with recombinant FIX under careful monitoring in intensive care unit considering of the inhibitor. After the slow endovenous infusion of 200 IU of recombinant FIX concentrate we stopped immediately the administration because the child presented cough, mild respiratory failure, tachycardia, tongue and lips oedema, lips cyanosis, diffuse vasodilatation, psychomotor agitation. He received also hydrocortisone, antihistaminic by intramuscular injection and oxygen by facial mask. The presence of inhibitor towards FIX and anaphylactic reaction occurrence preclude forever any replacement therapy with FIX both recombinant and plasmatic (PCC and/or aPCC). After this episode the patient needed another hospitalization for a tonsil bleeding with severe anaemization. We treat him with recombinant activated FVII (rFVIIa) first at the dosage of 270 mcg/kg in bolus, and after at the dosage of 90 mcg/kg every three hours for one day with complete bleeding remission. We encourage the careful monitoring of inhibitor towards FIX in haemophilic B patients especially with large FIX gene deletion. It could prevent severe anaphylactic reaction during replacement therapy. Considering the previous anaphylactic reaction, in this child rFVIIa represents the only therapeutic option for bleeding management.

Disclosures:

No relevant conflicts of interest to declare.
Essential Role for Mir-181a1/b1 In T-Cell Acute Lymphoblastic Leukemia
Authors: Fragoso Ana Rita , Mao Tin , Wang Song , Schaffert Steven , Min Hyeyoung , Pear Warren S. , Chen Chang-Zheng
Published: October 2019

Abstract 470

MiRNA-mediated gene regulation represents a fundamental layer of post-transcriptional control of gene expression with diverse functional roles in normal development and tumorigenesis. Whereas some studies have shown that over-expression of miRNA genes may contribute to cancer development and progression, it is yet to be rigorously tested by the loss-of-function genetic approaches whether miRNA genes are required for cancer development and maintenance in mice. Here we show that mir-181a1/b1 coordinates Notch and pre-TCR signals during normal thymocyte differentiation and plays an essential role in development and onset of T-cell acute lymphoblastic leukemia (T-ALL) induced by some Notch mutations. Using gain-of-function and loss-of-function approaches, we demonstrated that mir-181a1/b1 controls Notch and pre-TCR receptor signals during the early stages of T cell development in the thymus by repressing multiple negative regulators of both pathways, including Nrarp, PTPN-22, SHP2, DUSP5, and DUSP6. These results illustrate that a single miRNA can coordinate multiple signaling pathways by modulating the timing and strength of signaling at different stages. Intriguingly, synergistic signaling between Notch and pre-TCR pathways is necessary for the development of T-ALL, and miR-181 family miRNAs are aberrantly expressed in T-ALL patients. These observations raise the possibility that mir-181a1/b1 might contribute to the onset or maintenance of T-ALL by targeting similar pathways in tumor cells as it does in normal thymic progenitor cells. In support of this notion, we found that loss of mir-181a1/b1 significantly delayed the onset and development of T-ALL induced by intracellular domain of Notch1 (ICN1) and caused a 32% increase in the median survival time from 41 days to 54 days in T-ALL mice. Importantly, we noted that loss of mir-181a1/b1 more efficiently repressed the leukemogeneic potential of cells with lower levels of ICN1 expression, suggesting that mir-181a1/b1 may be more effective in inhibiting T-ALL development induced by a Notch mutant with weaker signal strength. Indeed, we demonstrated that loss of mir-181a1b1 essentially blocked T-ALL development induced by the weaker Notch mutant and dramatically decreased mortality from 60% to 10% in these T-ALL mice. Since human Notch mutations identified in T-ALL patients generally have weaker signaling strength and lower oncogenic potential than that of ICN1, our findings indicate that mir-181a1/b1 may play an essential role in development of normal thymic progenitors and Notch-induced T-ALL and may be targeted to treat T-ALL patients harboring Notch mutations.

Disclosures:

No relevant conflicts of interest to declare.
Incipient Release of Leukocytes After Allogeneic Hematopoietic Cell Transplantation Is Indicated by Increasing Serum Levels of Uric Acid Before Detection of First Leukocytes
Authors: Haen Sebastian P. , Eyb Vicky , Kanz Lothar , Bethge Wolfgang A.
Published: October 2019

Abstract 4701

Uric acid serves as an endogenous danger signal to the immune system upon the release from dying cells activating both innate and adaptive immunity. Multiple cytokines and danger signals are released into the blood modulating cellular and humoral immune responses during haematopoietic cell transplantation (HCT). Here, we investigated serum levels of uric acid during haematopoietic regeneration after allogeneic HCT to evaluate whether uric acid can be used as marker for hematopoietic engraftment indicating immunologic activity below the detection limit of differential blood counts (50/μ l). Differential blood counts and serum levels of uric acid, creatinine and urea were retrospectively analyzed in 47 consecutive patients (22 men and 25 women, median age 48 years, range 19 to 73 years) during 50 allogeneic HCT (three patients received a second transplantation due to recurrent disease). Serum levels of uric acid, creatinine and peripheral blood counts were determined daily. All patients received allopurinol for 5 days from the beginning of the conditioning which was discontinued afterwards. The mean nadir in uric acid levels was observed 13.4 days after the last dose of allopurinol (range 2 to 32 days) and 9.1 days after transplantation (range 2 to 21 days) irrespective of the conditioning regimen used. In all patients a significant decrease of uric acid serum levels after the transplantation was observed (Student's t-test, p < 0.001). A re-increase of uric acid was observed 1 to 10 days (mean 4.1 days, SD 2.4) before detection of white blood cells (leukocytes > 50/μ L) in 35 out of 50 transplantations (70%, group 1). In three cases (6%), the increase of uric acid levels was observed concomitantly with increasing leukocytes (group 2). In the remaining 12 cases, this elevation was only observed later (1 to 6 days after re-increase of leukocytes, mean 2.8 days, SD 1.5, group 3). The difference between group 1 and 3 was statistically significant (Student's t-test, p < 0.001). No correlation with patient age, conditioning regimen, HLA compatibility or additional medications could be observed. Therefore, a re-increase of uric acid can be indicative for incipient haematopoietic engraftment after allogeneic HCT and may serve as indicator for consumption of the ealiest differentiated cells in peripheral blood even before leukocytes can be detected.

Disclosures:

No relevant conflicts of interest to declare.
Studies on Preparation of Artesunate-Loaded mPEG-PLGA-Nanoparticles and Its Inhibition on K562 cells
Authors: Xiaoli Liu , Wang Shuang , Du Qingfeng , Xu Na , Yang Jun , Wang Yu , Zhang Jinfang , Xu Lulu , Li Xiang , Li Guofeng
Published: October 2019

Abstract 4712

The aim of the present work was to investigate the characterization of Artesunate-loaded methoxy poly (ethylene glyeol) -poly (lactic-co- glycolic acid) copolymer (mPEG-PLGA) nanoparticles and the anti-tumoral activity of the Art-Nps on human leukemia cells K562. The Artesunate (Art) poly (ethylene glyeol-lactic-co-glycolic acid) nanoparticles (Art-Nps) were prepared by modified-Spontaneous emulsion solvent diffusion method. The shape of the nanoparticles was observed by SEM. The mean diameter and the size distribution of nanoparticles were determined by laser light scattering. The drug loading efficiency, encapsulation rate and releasing behavior of Art -NPs in vitro were examined by HPLC. The effects of Art-Nps on the proliferation of K562 cells were studied by MTT assay and Hoechst staining. Artesunate loaded mPEG-PLGA nanoparticles were spheric with the mean size of (156.7±1.01)nm, zeta potential was -(26.23±1.86) mV, and the average drug loading and encapsulation efficiency were (14.5±0.2)% and (86.5±0.5)%, respectively. In vitro release behavior could be described by the Higuchi equation: Q=4.11t1/2+27.05, r=0.983. MTT assay showed different concentrations. different times of Art-Nps could inhibit the proliferation of K562 cells (P<0.01), and both have a synergistic effect (P=0.002), it showed that concentration was dependented with time, and the inhibition rate after 72h was exceeded to the the control group (P<0.05), it was showed the Art-Nps had sustained-release effect. Art-NPs of the cells (treated by 12.5μ g/m. 25μ g/ml. 50μ g/ml) resulted in significantly higher apoptosis than blank groups. The Art-NP obtained were characted with a small size and high drug loading and entrapment efficiency, in vitro release showed a good sustained-release nature, and it can inhibit the proliferation of human leukemia K562 cells in vitro, extending the time on leukemia cells. This study was provide an experimental basis for develop a new intravenous artesunate formulations.

Disclosures:

No relevant conflicts of interest to declare.
A Single Dose Filgastrim In the Treatment of Chemotherapy Induced Neutropenia
Authors: Jagarlamudi Kuppuswamy , Ahluwalia Meena , Patel Jay , Patel Arunbhai G.
Published: October 2019

Abstract 4774

Background:

Chemotherapy induced neutropenia (CIN) was a challenge to physicians until the discovery of growth factors. Neutropenia predisposes the patient to infection with increased morbidity, mortality, and cost of hospitalization. Not only does CIN leads to a delay in chemotherapy but also a dose reduction, which results in decreased efficacy and impact on survival. Filgrastim is a granulocyte colony stimulating factor that leads to the proliferation, differentiation and function of neutrophils. With an increase in the neutrophil count and function, Filgrastim reduces the incidence of neutropenia and the complications associated with febrile neutropenia (FN), making it possible to deliver chemotherapy on time, without dose adjustment. ASCO guidelines recommend that Filgastrim be given at 5 mcg/kg/day starting from days 1–3 after chemotherapy for patients with the risk of FN greater than 20% until the absolute neutrophil count is near-normal. It was recently reported that two doses of Filgrastim were comparable to higher number doses to prevent the nadir. We hypothesized that only a single dose is sufficient to prevent the nadir and delay in treatment.

Patients and Methods:

A retrospective analysis of chart data from all patients treated at The Brooklyn Hospital Center ambulatory clinic was reviewed after the initiation of chemotherapy. Data collected included the white cell count, neutrophil count, dates of chemotherapy and Filgrastim administration. Patients who received one dose of Filgastrim within 2–3 days, but not less than 24 hours prior to receiving subsequent chemotherapy comprised the study population. Data was analyzed for each individual cycle of chemotherapy.

Results:

The study population consisted of 11 patients with a total of 33 cycles of chemotherapy. Of the 11 patients, 9 had breast cancer, 1 had embryonal cancer, and 1 had colon cancer. Chemotherapy varied from Etoposide and Cisplatin, FOLFOX with Bevacizumab, and Adriamycin with Cyclophosphamid (AC) followed by Paclitaxel where the AC was given as a dose dense regimen. All the cycles had adequate response in neutrophil count to a single dose of Filgrastim when it was delivered within 2–3 days, but not before 24 hours of receiving chemotherapy. The mean absolute neutrophil count prior to Filgrastim was 0.8 compared to 6.0 (p=0.0001) after a single dose. There was no delay in treatment noticed as a result of the single dose of Filgrastim in all the 33 cycles.

Conclusion:

Filgrastim as a single dose is sufficient to treat CIN and prevent delay in subsequent treatment.

Disclosures:

No relevant conflicts of interest to declare.