bluuebunny/crossref_metadata_2025 huggingface dataset:
Multi-Course of Moderate Dose Cytosine Arabinoside and Fludarabin in the Consolidation Therapy of Patients with De Novo Acute Myeloid Leukemia
Authors: Qiu Huiying , Depei Wu , Sun Aining , Jin Zhengming , Miao Miao , Tang XiaoWen , Fu Chengcheng , Han Yue , He Guangsheng
Published: October 2019

Abstract 4251

To investigate the long-term outcome of multi-course of moderate dose cytosine arabinoside and fludarabin in the consolidation therapy of patients with de novo acute myeloid leukemia (AML). Sixty-seven consecutive de novo AML patients, 38 males, 29 females, including M1 (5 cases), M2a (34 cases), M4 (11 cases), M5 (13 cases), M6 (2 cases), unclassified AML (1 cases), HAL (1 case), were enrolled in this retrospective analysis between 2006 to 2011. Their median age was 32.28 years (range 13–64 years). Cytogenetic analysis showed that 27 patients with normal karyotype, 11 patients with t(8; 21), 4 patients with inv(16), 16 patients with other karyotype. Forty-two cases were classified as low and intermediate risk groups based on karyotype analysis and fusion gene detection. Thirty-eight cases and twenth-nine cases achieved complete remission (CR) after one course and two course induction chemotherapy (such as cytarabine with idarubicin or daunorubicin and mitoxantrone etc), respectively.

All patients received at least two-course consolidation therapy with moderate dose cytosine arabinoside (2g/m2) and fludarabin (30mg/m2) for 3 days (FA). Nineteen cases received two courses, 20 three courses, 20 four courses, 8 five courses and1 six courses. Quantitative RT-PCR and flow cytometry were used to detect minimal residual disease. Afterwards 3 patients underwent allogeneic stem cell transplantation. The median time for hematological recovery of neutrophils©ƒ500/microl and platelets©ƒ20,000/microl was 12 and 16 days, respectively. The median length of hospitalization was 18 days. Non-hematological toxicity consisted of mild gastrointestinal reaction and mild hepatic dysfunction, forty percent of patients experienced fever. There was no treatment-related mortality during consolidation. After follow up of 4–54 months, 10/19 (52.6%) cases were relapse-free survival (RFS) after two courses FA consolidation, 15/20 (75%) cases in three courses, 15/20 (75%) cases in four courses, respectively. 8 cases were all in RFS after five courses consolidation. 1 case was also in RFS after six courses FA consolidation. Three-year overall survival, event free survival and RFS was 67%, 67%, and 66%, respectively. Our results suggest that multi-course FA consolidation is effective in de novo AML patients with low-risk and moderate-risk diseases. To reduce the recurrence rate the number of courses should be increased. Non-hematological toxicity was mild and the toxicity is acceptable.

Disclosures:

No relevant conflicts of interest to declare.
Invasive Fungal Infections in Adults with Newly Diagnosed AML Undergoing Intensive Chemotherapy: Characterization, Risk Factors, and Outcomes in a Single Institution
Authors: Lu Kit , Neofytos Dionissios , Blackford Amanda , Seung Amy , Karp Judith E.
Published: October 2019

Abstract 4254

Background:

Invasive fungal infections (IFI) are a significant cause of morbidity and mortality in patients with acute myelogenous leukemia (AML) undergoing chemotherapy treatment. However, the epidemiology, risk factors, and outcomes of IFI in these patients have been poorly described.

Methods:

A single-center retrospective analysis was performed to study the epidemiology, risk factors, clinical outcomes, and mortality predictors of IFIs in AML patients undergoing intensive, multi-agent induction timed sequential therapy (TST) between January 2005 and June 2010. Newly diagnosed AML patients, with exception of acute promyelocytic leukemia, were studied. IFIs were defined using the European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) criteria.

Results:

254 consecutive patients (57% male; median age 54, range 20–78) were analyzed. 123 (48%) of patients had an IFI; of which, 15 patients (12%) had a proven candidal infection, and 108 patients (88%) had a mould infection (5 proven/probable (5%) and 103 (95%) possible mould infections). 237 (93%) patients received antifungal therapy during their treatment course (median day 8, range day -15 to 30). Of those, 63 (27%) received monotherapy (46% liposomal amphotericin, 44% voriconazole) and the rest received multiple antifungal agents.

Significant risk factors and trends for developing +IFI shown from univariate analyses are listed in Table 1. Prolonged neutropenia, duration of mucositis, and concurrent bacterial infections did not significantly affect the development of +IFI. Using multivariate analyses, mortality was impacted by patients’ baseline organ function (p=0.002 [1.3,3.1]), specifically, by bilirubin < 2 mg/dL (p=0.003 [1.38,4.57]) and creatinine < 1.5 mg/dL (p=0.01 [1.23,5.4]).

Patients with +IFI did not significantly impact overall survival. However, patients with candidal IFIs had a significantly lower overall survival compared to patients with mould IFIs and no IFI (HR 2.01 [1.06, 3.8]) (Figure 1). Candida colonization prior to or during the first week of chemotherapy, and development of mucositis were associated with the development of candidal IFIs (p=0.07).

Conclusion:

IFIs, particularly mould infections, remain a significant problem in patients with AML. Although overall survival did not appear to be significantly affected by mould infections, patients with candidal infections were more likely to die. Identification of risk factors for each type of IFIs may help to develop effective targeted preventive antifungal strategies.

Disclosures:

No relevant conflicts of interest to declare.
Clofarabine in Acute Myeloid Leukemia. the Spanish Experience
Authors: de la Fuente Adolfo , Deben Guillermo , Bethencourt Concha , Negri Silvia , Vilariño Dolores , Gonzalez Sonia , Olave Teresa , Martinez-Sanchez Pilar , Moscardo Federico , Caballero Mar , Fernandez Martinez Rosa , Bergua Juan , Gordillo Marina , Garcia Belmonte Daniel , Fernandez Fernandez Angeles , Solorzano Silvia , Santos Ana Belen , Roman Alejandro , Tormo Mar , Calabuig Marisa , Sastre Moral Jl , Lopez Lorenzo Jose Luis , Fernández-Abellán Pascual , Tomas Jose Francisco
Published: October 2019

Abstract 4279

Introduction:

Clofarabine (CLF) is a purine nucleoside analog approved by FDA and EMA in refractory pediatric ALL patients. Phase I and II studies have reported Clofarabine activity in AML.

Aims:

To evaluate the experience in Spain with (CLF) in the treatment of adult AML patients, analyzing effectiveness and toxicity profile.

Method:

This is a multicenter retrospective study including AML patients treated in Spain with CLF by compassionate use. Main points were complete remission as IWRv2003 criteria and toxicity as CTCAE v3.0 of NCI scale.

Result:

Between July 2007 ans April 2011 a total of 76 AML patients were treated with CLF based chemotherapy in Spain. We obtained clinical data from 64 cases. Median age at CLF treatment 52.4 years (18–77). Male/Female: 31/33. Previous Myelodisplastic syndrome 12 (18%). Cytogenetic data was available in 58 patients (90%), and 28 of them had high-risk cytogenetic.

CLF treatment: 56 patients received CLF as salvage therapy (23 in AML relapse and 33 in refractory disease), median previous lines 2 (0–7), were the remaining 8 patients were untreated patients. CLF was administered in combination with AraC in 94% patients and the 91% received a five days schedule. The most frequents CLF dose/day were: 40 mg/m2/day in 39 patients, 30 mg/m2/day in 11 patients, and 20 mg/m2/day in 9 patients.

Response and outcome: Nineteen (35.8%) patients achieved complete remission and 64.1% had resistant disease. Mean survival time 14.3 ± 1.4 months.

The statistical analysis shows a significant difference in the CR rate between first line therapy (CR 82%) and savage therapy in relapsed (CR 45%) and salvage therapy in refractory (CR 18%), (p0.012). Neither adverse cytogenetics nor previous MDS did influence CR. (p0.57 and p0.53 respectively).

Toxicity: All patients presented grade IV hematological toxicity with grade IV neuthopenia, grade IV trombopenia and transfusion depend anemia. The incidence of extra hematological toxic effects were low, creatinine >3 mg/dL: 4 cases (6%); bilirrubine > 3mg/dL: 9 cases (14%). Eleven (17%) patients died during Induction.

Conclusions(opcion1):

The revised experience in Spain with CLF as compassionate used in AML confirms the effectiveness of Clofarabine as a salvage regimen. Of note was that CLF could potentially overcome some adverse known factors as cytogenetics.

Disclosures:

Off Label Use: Clofarabine in AML is an off-label use.
Wogonoside Exerts Anti-Leukemia of AML Cells Via Restricting Phospholipid Scramblase 1 Gene Expression and Promoting Its Translocation Into Nuclear
Authors: Chen Yan , Chen Bao-An , Guo Qing-long
Published: October 2019

Abstract 4282

Objective:

To evaluate the antileukemic effect of wogonoside and reveal the underlying mechanism.

Method:

In this study trypan blue dye exclusion assay, MTT assay, and soft agar colony formation assay were used to analysis growth inhibition of wogonoside the on AML (acute human promyelocytic) cell lines. Propidium iodide (PI)-staining and cell cycle-regulatory proteins detecting by western blots were applied to exam whether wogonoside could induce cell cycle arrest. Then a series of experiment were used to assess the ability of wogonoside to overcome the AML associated differentiation block, by using Giemsa staining, Nitroblue tetrazolium (NBT) reduction assay, and cell-surface differentiation antigens expression analysis. Real time PCR, western blots, cycloheximide inhibition test and RNA interference, nuclear and cytoplasmic fractionation, immunofluorescent staining were used to investigate the underlying mechanism. In this point we mainly focus that wogonoside exerts antileukemic by modulating of PLSCR1 gene expression, as well as influence its subcellular localization to play a role in regulating gene transcription.

Result:

It was demonstrated that wogonoside have the capacity to decrease the growth of myeloid cell lines by induction of G0/1 phase cell cycle arrest and differentiation. This effect is mediated by the increasing in mRNA and up-regulating protein expression of phospholipids scramblase 1 (PLSCR1). Meanwhile wogonoside promoted PLSCR1 traffic into the nucleus, which let PLSCR1 to play a role in regulating cell cycle and differentiation-related genes transcription including p21, p27, c-myc and IP3R1.

Conclusion:

Wogonoside induced AML cell lines to undergo differentiation and G1 phase arrest by restricting phospholipid scramblase 1 gene expression and promoting its translocation into nuclear.

Disclosures:

No relevant conflicts of interest to declare.
The Analysis of Fe3O4 Nanoparticles Combined with Daunorubicin for Neurotoxicity of Rat Brain In Vivo
Authors: Xu Peipei , Chen Bao-An , Wang Xue-mei , Li Jingyuan , Cai Xiao-hui , Jiang Hui
Published: October 2019

Abstract 4287

Introduction□F

The safety of these nano-systems for human body should be carefully studied for the potential clinical application. Here in this report, we firstly study the effect of anticancer drug daunorubicin (DNR) and Fe3O4 nanoparticles (NPs) for the neurotoxicity of rat brain. The microdialysis experiment gave the precise and real-time method to investigate the effect.

Method:

SD rats(240–250 g) were randomly assigned to three groups containing three rats in each group. Microdialysis Probes were deployed corpus striatum. Then the animals were solitary breed to recovery the wound for about 24 h. The different drugs systems were administered by vena caudalis injection. Amino acids were measured in dialysates by HPLC.

Results:

The results indicated that the anticancer drug DNR had the serious bad effect for the rat brain to cause the neurotoxicity by influencing the concentration changes of amino acids in which some excitatory amino acids (such as Asp) and the inhibiting amino acid (such as Gly) especially decreased and the inhibiting amino acid GABA increased. For the system of nanocomposites for DNR conjugated with Fe3O4 NPs, the side effect of DNR for these amino acids was visibly cut down, and the time for DNR to affect the side neurotoxicity for the rat brain was shorten.

Conclusion□F

So these nanocomposites for DNR conjugated with Fe3O4 NPs had the better biocompatibility and biosecurity in the application of cancer therapy in vitro and in vivo. These nanocomposites system had the potential prospect to be further studied and utilized in the clinic therapy.

Disclosures:

No relevant conflicts of interest to declare.
Should Generic Versions of Low Molecular Weight Heparins Be Considered As Generic or Biosimilar Drugs. A Regulatory Dilemma
Authors: Fareed Jawed , Jeske Walter , Hoppensteadt Debra , Wahi Rakesh , Hull Russell D
Published: October 2019

Abstract 4328

The patent protections for the low molecular weight heparin (LMWHs) enoxaparin, dalteparin and tinzaparin have expired and their generic respective versions are available. Despite widespread clinical use the global regulatory pathways to develop and approve these products remain unclear. In South America and Southeast Asia generic versions of LMWHs are available without adequate regulatory oversight. The EMEA has considered these agents as biosimilars and require both pharmaceutical and clinical data for approval. Unlike the EMEA, the US FDA requires data on the basic, chemical, and pharmacologic profiles without any clinical validation. In July 2010, the FDA approved a generic version of enoxaparin for all of the clinical indications for which the branded enoxaparin is approved. On the other hand, Initially the FDA had approved the different branded LMWHs as distinct drug entities, requiring clinical trials for each individual clinical indication. Generic enoxaparins from various manufacturers are marketed under different trade names even when they are manufactured by the same company. Although in the pharmacopeial assays these agents exhibit comparability, in the pharmacodynamic studies assay based differences have been reported. In a study conducted on two batches of Sandoz’s enoxaparin and Lovenox®, while no differences were noted in the molecular profile and anti-FXa activity, significant differences were noted in other assays such as thrombin generation inhibition and PF4 titration. Moreover, pharmacodynamic differences in primates after sc administration were also noted which were more obvious in the release of tissue factor pathway inhibitor and thrombin activatable fibrinolysis inhibitor assays (TAFI).As the manufacturing process of these agents may differ from manufacturer to manufacturer it is likely that certain compositional variations are not detectable by chemical methods. Thus the requirement of clinical trials to assure the safety and efficacy of these agents is an important consideration. Of concern is the disharmonization of pathways between different regulatory bodies which contribute to the complexity of this issue. It is proposed that the EMEA, FDA and other regulatory bodies develop some common global guidelines for the rationale development of these drugs.

Disclosures:

No relevant conflicts of interest to declare.
Study of Octaplex Dosing Accuracy: An In Vitro Analysis
Authors: Patriquin Christopher Jordan , Chin-Yee Ian H. , Kovacs Michael J. , Lazo-Langner Alejandro , Keeney Michael , Hsia Cyrus C.
Published: October 2019

Abstract 4335

Background:

Supratherapeutic INRs are common with warfarin therapy, and increase serious bleeding risk. Octaplex is a prothrombin complex concentrates (PCC) that is recommended for urgent warfarin reversal. However, disagreement exists regarding proper dosing strategies (fixed versus weight-based).

Objectives:

We sought to measure the in vitro effect of Octaplex on INR and factor levels, and to characterize this relationship.

Methods:

Plasma samples from eligible patients on warfarin with stable INRs for ≥4 weeks were collected. Plasma volumes were calculated to approximate 1000, 2000 and 3000IU doses of Octaplex and these were added to the samples. INR and factor levels were measured pre- and post-Octaplex.

Results:

Twenty-three of the 30 subjects enrolled had complete data for analysis. INRs corrected <1.5 in all samples post-1000IU, and decreased further with subsequent doses (p<0.001). Similar changes occurred in factors II, VII, and X (p<0.01). Linear correlations were seen between INR and factors II, VII and X. Factor IX did not increase incrementally or show a correlation with INRs. Weight-based dosing was then estimated. All INRs were <1.2 (0.9–1.2), and factor levels >0.50IU for II, VII and X (0.96–1.52, 0.51–1.45 and 0.81–1.38, respectively). Factor IX did not uniformly correct >0.50IU (0.31–1.31).

Conclusion:

We confirmed in vitro that 1000IU of Octaplex was able to correct INR to <1.5 but factor activity for II, VII and X was not uniformly >0.50IU until 2000IU, and >1.00IU not until 3000IU. Our study suggests that INR correction alone may not appropriately reflect sufficient factor activity, and lends support for weight-based PCC dosing.

Disclosures:

No relevant conflicts of interest to declare.
ABO Genotype-Phenotype Discrepancy Due to Tri-Allelic ABO Genotypes and Its Resolution by Cloning-Sequencing and Chimerism Analysis
Authors: Lee Min-Young , Park Tae Sung , Lee Jeong Nyeo , Kim Hye Ran , Oh Seung Hwan
Published: October 2019

Abstract 4337

We investigated the molecular genetic basis of an ABO blood group discrepancy found incidentally in an individual with no transfusion or transplantation history. During routine preoperative blood grouping, red blood cells (RBCs) had a weak or mixed field reaction (double populations in a gel column) with anti-A, while serum typing showed the anti-B antibody. We sent the sample to a reference laboratory for ABO genotyping using sequencing analysis, and her genotype was reported to have O/O alleles. To resolve the genotype-phenotype discrepancy, we repeated the direct sequencing analysis and finally found a heterozygous insertion/deletion (indel) on exon 6, involving two major alleles and one minor allele. With cloning and sequencing analysis, we isolated the O01, O02, and A102 alleles, respectively. In addition, tri-allelic or tetra-allelic patterns in short tandem repeat (STR) loci were noted. Chimerism is rare, but it can cause genotype-phenotype discrepancies in blood grouping, parentage, or other genetic laboratory tests. The transfusion of chimeric RBCs was even reported to induce acute intravascular hemolysis. It is very difficult to correctly find heterozygous peaks without assuming their existence, especially when caused by a chimera. Consequently, separation by cloning or enrichment of minor alleles such as using the cold polymerase chain reaction is sometimes needed. A familial study of STR markers could also be a confirmatory test. Here, we report an ABO genotype-phenotype discrepancy due to tri-allelic ABO genotypes and its resolution by cloning sequencing and STR analysis.

By cloning-sequencing analysis, the two major alleles O01 and O02 and one minor allele A102 were separated. A blank means the same base as the reference sequence. Bases in parentheses indicate minor peaks.

Disclosures:

No relevant conflicts of interest to declare.
Increased Levels of PTX3 in Human Plasma in Septic Patients with DIC
Authors: Kawasugi Kazuo , Yamamoto Tadashi , Shirasaki Ryosuke , Tashiro Haruko , Shirafuji Naoki
Published: October 2019

Abstract 4347

Sepsis is a life-threatening condition that is characterized by a whole-body inflammatory state (called a systemic inflammatory response syndrome, SIRS). Long pentraxin PTX3 is an inflammatory mediator and a component of the humoral arm of innate immunity produced by neutrophils, macrophages, myeloid dendritic and endothelial cells. During sepsis a massive inflammatory activation and coagulation/fibrinolysis dysfunction occur. However, little is known about PTX3 in septic patients with DIC. Therefore, we measured PTX3s in the plasma from septic patients with DIC (n=20). Also, we investigated PTX3 in the plasma from septic patients without DIC (n=8) and acute promyelocytic leukemia (APL)-induced DIC (n=5). PTX3 in the plasma from human were measured using enzyme-linked immunosorbent assay (ELISA) kits (Perseus Proteomics Inc, Japan).

The thrombin antithrombin complexes (TAT) levels were higher in both DIC patients as reported by others. We detected high levels of PTX3 in the plasma of all septic patients with DIC. Also, plasma levels of PTX3 were positive in septic patients without DIC. However, plasma levels of PTX3 were significantly higher in septic patients with DIC than in septic patients without DIC. Plasma levels of PTX3 ware significantly correlated with severity of septic DIC (platelet count and TAT level). We did not find any difference plasma levels of PTX3 in the APL patients with DIC. Moreover, high-mobility group box 1 (HMGB1) concentrations in the human plasma were determined using ELISA kit (Shino-Test, Japan). Plasma levels of HMBG1 were positive for 14 cases in 20 septic patients with DIC. Also, we did not find any difference plasma levels of HMGB1 in the APL patients with DIC.

These results suggest that assess of PTX3 and HMBG1 in the plasma may be helpful in the making the diagnosis in septic patients with DIC. However, PTX3 seemed to be a better biomarker than HMGB1. It appears that PTX3 may contribute to the severity of septic DIC.

Disclosures:

No relevant conflicts of interest to declare.
Detection of Circulating ADAMTS13 Immune Complexes by Antibody Class-Specific ELISA
Authors: Plaimauer Barbara , Ferrari Silvia , Gruber Bernadette , Rottensteiner Hanspeter , Turecek Peter L , Ehrlich Hartmut J. , Scheiflinger Friedrich
Published: October 2019

Abstract 4361

A severe functional deficiency of the von Willebrand factor (VWF) cleaving protease ADAMTS13 results in the accumulation of uncleaved, highly adhesive VWF multimers in plasma leading to increased platelet aggregation and thrombus formation in the microcirculation. Anti-ADAMTS13 autoantibodies that either neutralize ADAMT13’s activity or enhance protein clearance are the major cause of severe ADAMTS13 deficiency in acquired thrombotic thrombocytopenic purpura (TTP). Circulating anti-ADAMTS13 immune complexes that may play a role in disease pathogenesis have been described in plasma of TTP patients. We have developed a new ELISA assay to specifically determine the levels of ADAMTS13-specific circulating immune complexes (CIC) in which the antigen portion is immobilized by a polyclonal anti-ADAMTS13 IgG and the immunoglobulin component detected by a class-specific antibody. An inverse correlation between levels of circulating anti-ADAMTS13 autoantibodies and ADAMTS13-specific CIC has been observed in plasma of acquired TTP patients. The correlation between free anti-ADAMTS13 titer and the level of CIC and insight into the immunoglobulin (sub) classes involved may provide valuable prognostic information about disease severity and progression, and aid patient treatment monitoring.

Disclosures:

Plaimauer: Baxter Innovations GmbH: Employment. Ferrari:Baxter Innovations GmbH: Employment. Gruber:Baxter Innovations GmbH: Employment. Rottensteiner:Baxter Innovations GmbH: Employment. Turecek:Baxter Innovations GmbH: Employment. Ehrlich:Baxter Innovations GmbH: Employment. Scheiflinger:Baxter Innovations GmbH: Employment.
Neurotrophic Bone Marrow Cellular Nests Prevent Spinal Motoneuron Degeneration in Amyotrophic Lateral Sclerosis Patients
Authors: Blanquer Blanquer Miguel , Iniesta Francisca , Gómez Espuch Joaquín , Ramón Villaverde , Pérez Espejo Miguel Ángel , Ruíz López Francisco José , García Santos José María , Bleda Patricia , Izura Virginia , Sáez María Vicenta , de Mingo Pedro , Vivancos Laura , Carles Rafael , Majado Maria Juliana , Sánchez Salinas Andrés , Martínez-Lage Juan Francisco , Martínez Salvador , Moraleda José M.
Published: October 2019

Abstract 4393

Background:

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease, characterised by loss of motoneurons (mns), and it has no cure. Cell therapy has neurotrophic effects in animal models and has been proposed as a disease-modifying treatment. Our aim was twofold: firstly, to assess the safety of intraspinal infusion of bone marrow mononuclear cells (BMNC) and, ultimately, to look for histopathological signs of cellular neurotrophism.

Methods:

We conducted an open single arm phase I trial. After six months observation, autologous BMNC were infused into the posterior spinal cord funiculus. Safety was the primary endpoint and was defined as the absence of serious transplant-related adverse events. In addition, forced vital capacity (FVC) and ALS-FRS, MRC and Norris scales were assessed six and three months prior to the transplant and quarterly afterwards for one year. Pathological studies were performed in case of death.

Findings:

Eleven patients were included. We did not observe any severe transplant-related adverse event but there were 43 non-severe events. Twenty-two (51%) resolved in ≤2 weeks and only four were still present at the end of follow-up. All were CTCAE grade ≤2. No acceleration in the rate of decline of FVC, ALS-FRS, Norris or MRC scales was observed. Four patients died on days 359, 378, 808 and 1058 postransplant for reasons unrelated to the procedure. Spinal cord pathological analysis showed a greater number of montoneurons in the treated compared to the untreated segments (4.2+/−0.8 mns/section and 0.9+/−0.3 mns/section, respectively). In the treated segments, motoneurons were surrounded by CD90+ cells and did not show degenerative ubiquitin deposits.

Interpretation:

This clinical trial confirms not only the safety of intraspinal infusion of autologous BMNC in ALS patients but also provides evidence of their neurotrophic activity.

Disclosures:

No relevant conflicts of interest to declare.
OPTIMIZATION of the Cultivation of Mesenchymal STEM CELLS for CLINICAL USE
Authors: de Souza Valim Vanessa , Oliveira Fernanda , Lima da Silva Maria Aparecida , Amorin Bruna , Moraes Lauro , Pezzi Annelize , Dahmer Alice , Carvalho Regina , Lemos Natália , Baggio Letícia , Kersting Nathália , Sehn Filipi , Melchiades Gabriela , Daudt Liane , Silla Lucia
Published: October 2019

Abstract 4394

Introduction and objectives:

Studies with mesenchymal stem cells (MSCs) have shown its benefits in hematology, mainly for Graft-versus-host disease (Lancet 371:1579–86, 2008), with three unsettled matters: (1)MSCs expansion in medium supplemented with Fetal Calf Serum (FCS) and its risk of xenoreaction (Blood 89:776–9, 1997); (2)The optimal number of cells needed for therapy is not yet defined, but there is an empirical indication for 2×106células/Kg with the need to optimize expansion, number and time wise; and (3)the utilization of third party donors. This study was designed to determine the superiority or no-inferiority of the Platelet Lysates (PL) over FCS on the expansion of MSC, the optimal cell plating density and days between each pass, and to investigate if in our conditions total nucleated cells (TNC) obtained from the washouts of HSCT explants can expand to be used at clinical grade.

Methods and Results:

TNC were removed from the filters and bags used in the HSCT (Cytotherapy 11:403–13, 2009) and after the first passage were plated in different concentrations (2000/cm2, 3000/cm2, 4000/cm2, 5000/cm2, 6000/cm2 and 7000/cm2) with 10% FBS or 10% PL, and the number of days reach 80% of confluence was observe (Transfusion, 49:2680–5, 2009). Next, cultures with the same plating density were fed either with 10% PL or 10% FCS and were trypsinized after seven days and counted to analyze how much they have grown in that time period. And finally, cultures were allowed to growth up to Passage 3 (P3) to test the ability to obtain clinical grade number of cells. The proliferation of mesenchymal stem cells in the presence of PL and SFB was averaged 11.88 and 2.5 times, respectively, in a period of 7 days (p = 0.005). The highest concentration of plating cells using PL, took less time (6 days) to reach confluence as compared with the three lower (7.55 to 8.55 days) (p = 0.005), and at P3 with PL we obtained from 10×109 up to 10 × 1011 cells.

Conclusion:

This study suggests that the PL is the best choice as a supplement to expand MSC, and allow the proliferation of a sufficient number of MSC at P3 for clinical use obtained from the washouts of HSCT explants.

Disclosures:

No relevant conflicts of interest to declare.
Distinctive Spectra of Cytogenetic Alterations in Over Five Hundred East Asian Patients with Acute Leukemia
Authors: Choi Hyun-Jung , Kim Hye Ran , Shin Myung-Geun , Shin Jong-Hee , Park Bock-Hee , Ryang Dong-Wook , Suh Soon-Pal
Published: October 2019

Abstract 4408

Background:

Racial difference of spectra of chromosomal aberrations in various hematological malignancies was previously reported between Asian and western countries. Therefore, characterization of the cytogenetic profile is still a matter of interests for identifying cytogenetic risk factors in the current risk-adapted chemotherapy. The aim of this study was to investigate the spectrum of chromosomal aberrations in East Asian (Korean) acute leukemia patients, and to propose a panel of leukemic fusion genes suitable for the development of new molecular detection systems.

Patients and Methods:

We prospectively analyzed bone marrow samples from 502 patients with acute leukemia referred for screening leukemic fusion genes by simultaneous analysis of conventional cytogenetics, FISH and multiplex RT-PCR system in Chonnam National University Hwasun Hospital (Hwasun, Korea) for last 5 years.

Results:

A total of 334 patients were diagnosed with AML, 155 with ALL and 13 with mixed phenotype acute leukemia. Twenty types in 28 fusion genes were detected in 42.4% of the total patients by multiplex RT-PCR system and in 33.9% by conventional cytogenetics including FISH. In the group of AML, the common fusion genes were 57 PML/RARa, 38 AML1/MGT8, 12 CBFb/MYH11, 11 MLL1, 5 BCR/ABL1, SET/CAN, PLZF/RARA and TLS/ERG in 2 cases each, and one of each AML1/MDS1, TEL/ABL and TEL/MN1. In the group of ALL, the following fusion genes were detected: 33 BCR/ABL1, 22 TEL/AML1, 10 MLL1, 7 E2A/PBX, 2 SET/CAN and SIL/TAL, one TEL/ABL. In addition, cytogenetically cryptic translocations were detected in 14.8% patients with normal karyotype or numerical aberrations by conventional cytogenetics. Among 288 patients with negative results by multiplex RT-PCR system, one child with early pre-B-ALL harbored MLL1/AF4, and 8 patients had 3 types of clinically significant chromosomal aberrations such as t(3;3)(q21;q26.2), t(8;14)(p24.1;q32) and i(17)(q10), which were all confirmed by conventional cytogenetics.

Conclusions:

The current study demonstrated the spectrum and frequency of chromosomal aberrations in Korean patients with acute leukemia, which are different from Caucasian data. Also these results may offer important implications for the development of new multiplex molecular detection system.

Disclosures:

No relevant conflicts of interest to declare.
Dasatinib As a Late Second Line Therapy in Chronic Myeloid Leukemia Patients: A Retrospective Study From a Tertiary Care Centre in India
Authors: Seth Tulika , Sharma Vandana , Sharma Sanjeev , Mahapatra Manoranjan , Mishra Pravas , Kumar Rajat , Pati Hara prasad
Published: October 2019

Abstract 4438

Introduction-

Chronic myeloid leukemia in Indian patients manifests a decade earlier than reported in the West. However, patients present later to the hospital, with advanced disease as evinced by large spleen size, fever and other symptoms and a high Sokal score. The cost of medicines and lack of health insurance means that most will receive Imatinib mesylate (Glivec) from the GIPAP program only, or locally produced Imatinib. The usual option for imatinib failure is to increase the dose to 600 or 800mg. Switching patients to second line therapy is difficult due to financial constraints; patients who fail Imatinib are offered allogeneic stem cell transplantation.

Methods-

We present a single-institution retrospective data of Dasatinib as second line therapy in CML patients resistant to imatinib. In a total of 1200 CML patients followed in our clinic, 24 patients were treated with dasatinib over a period of 4 years. 17 of whom were in chronic phase (switched after failure of Interferon and Imatinib -2, not in CHR at time of switch- 4), accelerated phase-5 and blast crisis-2 patients, at the time of starting dasatinib. They either refused transplant, did not have donor or had co-morbid factors (3-diabetes mellitus and CVD, 1- chronic renal disease along with diabetes) which made transplant difficult.

Results-

The median age of patients was 45 years (26–65 years) with 12 males and 12 females. They had received imatinib for a median of 50 months (20–98 months) before starting dasatinib. After a median follow up of 12 months (6.6 – 39.6 months), there was 2 deaths (CML-BC-1, CML-AP-1). Non-hematologic toxicity was generally mild to moderate (grades 1 or 2); dyspnea, gastrointestinal disorders (diarrhea, nausea) and fluid retention were seen. Pleural effusion occurred in two patients.

Conclusions-

Toxicities with Dasatinib were generally reversible and could be managed effectively with dose adjustments. Our limited experience showed this to be effective and well tolerated even in late failure patients with high disease burden.

Disclosures:

No relevant conflicts of interest to declare.
Treatment of Consecutive Patients with Chronic Myeloid Leukemia In the Cooperating Centers From the Slovak Republic After 2000 – A Report From the Population Based CAMELIA Registry
Authors: Demeckova Eva , Mistrik Martin , Demitrovicova Ludmila , Chudej Juraj , Markuliak Imrich , Tothova Elena , Muzik Jan , Dusek Ladislav
Published: October 2019

Abstract 4439

Background:

Most results on the treatment of chronic myeloid leukemia (CML) with imatinib were obtained from clinical trials that may differ from the routine practice. We report the results of treatment of consecutive CML patients at ten major centers during 2000–2010.

Patients and methods:

Data reporting was retrospective in 2000– 2004 and prospective from 2005 on. A total of 295 patients (137 women and 158 men; median age 49 [range, 15–81]) with Ph+ CML were registered. The median follow-up was 45.4 months (0–113,5).

Results:

Most patients were treated with first- (169; 57.28%) or second-line (84; 28.5%) imatinib, part of patients underwent allogeneic hematopoietic stem cell transplantation (AHSCT) (28; 9,5%), but 4,7 % were treated with other modalities (14 patients; median age 66 [range, 32–83]). The probability of overall survival (OS) according to Kaplan and Meier at five years was 88.9%, 77.5% and 68.7% for chronic phase patients treated with first-, second-line imatinib and first-line AHSCT, respectively, but only 25.2% for patients receiving other modalities. OS was dependent on the disease phase and Sokal, Hasford and EBMT risk scores (p< 0.001; each). Only 46.2% of deaths in patients treated with other modalities were attributable to CML. Elderly patients over 65 years achieved similar response rates and progression-free survival as the younger. There was a trend for inferior results of AHSCT performed after failure of imatinib (p=0.075), probably due to differences in EBMT risk scores (p< 0.001).

Conclusions:

Ability to achieve results comparable to previous clinical studies in our CML cohort was influenced by centralized care. Decisions not to initiate imatinib or to delay AHSCT may have a negative impact on OS, but comorbidities may limit the treatment potential of imatinib in the elderly.

Disclosures:

No relevant conflicts of interest to declare.
Efficacy and Safety of Dasatinib in Chronic Myeloid Leukemia Patients in Chronic Phase (CML-CP) with Imatinib Resistance or Intolerance
Authors: Akagi Tomoaki , Murai Kazunori , Shimosegawa Kenji , Ishizawa Kenichi , Sugawara Tomohiro , Yokoyama Hisayuki , Noji Hideyoshi , Ogawa Kazuei , Tazima Katsusi , Ooba Koji , Sakamoto Junichi , Koumei Kubo , Sakamaki Hisashi , Harigae Hideo , Ishida Yoji
Published: October 2019

Abstract 4444

Background:

Dasatinib is highly potent BCR-ABL inhibitor, with 325-fold higher potency than imatinib and a 16-fold higher potency than nilotinib in vitro. Across a series of phase II and III trials with more than 2 years of follow-up, dasatinib has demonstrated durable efficacy in patients with CML following resistance or intolerance to imatinib. We conducted a phase II study to evaluate the efficacy and safety of dasatinib in patients with imatinib resistance or intolerance in the Japanese population.

Methods:

54 CML- CP patients (median age 63.5 years) with resistance, intolerance or a suboptimal response to imatinib were submitted to dasatinib 100mg once daily. Efficacy and safety were assessed using rates of major molecular response (MMR)/ complete molecular response (CMR) or drug-related adverse events (AEs) respectively. All analyses were based on the intension-treat principle.

Results:

Until now, the results of 32 patients, who passed more than 12 months, have been analysed. The median time since of prior imatinib therapy was 19.2 months with 57.4 % being imatinib-resistant and 42.6 % imatinib-intolerant. The incidence of MMR (primary endopoint) and complete molecular response at 12 months was 59.3 % (95% confidence interval, 45.0 – 72.4 %), and 24.1 % patients, respectively. Progression to the accelerated or blastic phase had not been observed. All patients with baseline samples tested for mutations, and different mutations were not observed. Non-hematological AEs were mostly grade 1/2. Grade 3/4 non-hematological AEs were infrequent, including increased AST (1.9%), and increased creatinine (1.9%), respectively. Grade 1/2 fluid retention adverse events were shown in 10 patients, including 2 patients with edema, and 9 patients with pleural effusion. Grade 3/4 hematological toxicities including anemia (1.9%), neutropenia (14.8%), and thrombocytopenia (9.3 %) were shown, and 13 patients have permanently discontinued treatment due to adverse effects.

Conclusions:

Dasatinib treatment results in high rate of molecular response in CML-CP patients with resistance or intolerance to imatinib, and approximately 60% of evaluated patients who switched to dasatinib achieved MMR at 12 months. Importantly, among all patients who achieved molecular response, 24.1 % patients achieved CMR. Dasatinib at a dose of 100 mg once daily was generally well-tolerated, and had better efficacy for patients with CML-CP post imatinib failure.

Disclosures:

No relevant conflicts of interest to declare.
Long-Term Effects of Kidney Shielding During Total Body Irradiation: A Single Institution Experience
Authors: Kobayashi Takeshi , Kakihana Kazuhiko , Ohashi Kazuteru , Sakamaki Hisashi , Karasawa Katsuyuki , Akiyama Hideki
Published: October 2019

Abstract 4475

Introduction:

Long-term follow-up of patients undergoing allogeneic hematopoietic stem-cell transplantation (HSCT) revealed a high incidence of chronic kidney disease (CKD), prompting us to introduce kidney shielding during total body irradiation (TBI) (kidney dose of 10 Gy).

Subjects and methods:

To evaluate the effects of the shielding maneuver, we evaluated patients with acute lymphoblastic leukemia in first complete remission who had received allogeneic transplantation between 1990 and 2008. All patients had the same preparative regimen, with the exception of kidney shielding, which started in March 1999.

Result:

Seventy-two patients were included, 21 without shielding (GNS) and 51 with shielding (GS). The historical comparison led to more high-risk patients in the GS with regard to graft-versus-host disease. The incidence of acute GVHD graded more than II was higher in the GS (p=0.015), but this trend was not observed for chronic GVHD. The incidence of acute kidney injury diagnosed according to the RIFLE criteria was very similar in the two groups, whereas the incidence of CKD at 2 and 5 years after transplantation was significantly lower in the GS group (p=0.046). Five patients in the GNS developed renal failure that eventually required dialysis (9 to 17 years after transplantation), whereas no patients in the GS have required dialysis to date. The incidence of relapse demonstrated no significant differences between the two groups, irrespective of Philadelphia chromosomal status. There were no cases of relapse that started in the kidney or surrounding tissue. Overall survival was lower in the GS, but the difference was not statistically significant (p=0.286).

Conclusion:

Protective shielding during TBI is able to suppress the incidence of CKD at 5 years after transplantation, without any significant effect on the risk of relapse.

Disclosures:

No relevant conflicts of interest to declare.
Characterization Of An Acquired IgG Autoantibody To Bβ and γ Chains Of Fibrinogen Resulting In Delayed Fibrin Polymerization and Severe Bleeding
Authors: Lulla Premal , Dasgupta Swapan , Rice Lawrence , McCarthy John J. , Thiagarajan Perumal
Published: October 2019

A 88 year-old woman was referred to our center for 3 weeks of gastrointestinal bleeding and a spontaneous right rectus femoris hematoma. The prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT) and reptilase time (RT) were markedly prolonged (Table 1). Assays for coagulation factors V, VIII, IX and X were normal. A fibrinogen concentration of < 50 mg/dl was measured by Clauss method while fibrinogen concentration measured by radial immunodiffusion was 180 mg/dl. Following infusions of cryoprecipitate there was accelerated clearance of fibrinogen as measured by the Clauss method. An inhibitor of fibrinogen function was suspected.Table 1TestTime (sec)1:1 mix (sec)Normal Range (sec)PT>902212-15aPTT70.73923-36TT>120>12015-19Reptilase Time>120>12014.8-20.4

When the patient’s plasma was tested towards immobilized fibrinogen in ELISA plates, a concentration dependent specific binding of IgG was seen while under similar conditions control IgG had no significant binding (Figure 1). The IgG antibody was polyclonal comprising of both kappa and lambda subtypes.

The IgG fraction was purified on a Protein A column and the isolated IgG fraction prolonged the thrombin time of normal plasma in a concentration dependent manner. The effect of the isolated IgG fraction on fibrin polymerization was measured by monitoring the absorbance at λ390 in a spectrophotometer. The IgG inhibited fibrin polymerization in a concentration dependent manner. Control IgGs had no significant effect on polymerization under these conditions (Figure 2).

The immunologic specificity of antibody was tested in immunoblots of fibrinogen subjected to SDS-PAGE. The patients IgG reacted with fibrinogen in unreduced buffers. Under reducing conditions, the IgG reacted with Bβ and γ chains of fibrinogen while the control IgG did not (Figure 3).

The patient was treated with rituximab and steroids. After four, weekly doses of rituximab, fibrinogen levels by Clauss method normalized (450 mg/dl). There was no detectable antibody to fibrinogen and no further bleeding was noted.

Conclusion

This is a description of a rare case of a polyclonal autoantibody to Bβ and γ chains of fibrinogen that inhibited fibrin polymerization leading to a severe bleeding diathesis.

Disclosures:

No relevant conflicts of interest to declare.
Thrombomodulin Alfa Treatment For Disseminated Intravascular Coagulation In Acute Promyelocytic Leukemia: A Retrospective Analysis Of The Open-Label, Multicenter Study Cohort
Authors: Matsushita Tadashi , Watanabe Jyunichi , Honda Goichi , Mimuro Jun , Takahashi Hoyu , Tsuji Hajime , Eguchi Yutaka , Kitajima Isao , Sakata Yoichi
Published: October 2019

Background

Patients with acute promyelocytic leukemia (APL) patients present disseminated intravascular coagulation (DIC) that can result in life-threatening hemorrhagic complications. In 2008, the anticoagulant thrombomodulin alfa (TM-α, recombinant human soluble thrombomodulin) was approved for the treatment of DIC in Japan. After commercial launch, only small case series have been described in APL patients, and data from larger patients cohort are awaited. We accessed the clinical safety and beneficial effects of TM-α treatment in DIC patients with APL, by evaluating data on the open-label, multicenter, post-marketing surveillance study cohort.

Patients and Methods

A retrospective evaluation was carried out on a cohort of 172 patients with APL from the open-label, multicenter, post-marketing surveillance study of TM-α in Japan. Of all 172 patients, 31 were relapse/refractory APL patients and 141 were newly diagnosed. The newly diagnosed APL patients were generally received all-trans retinoic acid (ATRA) and anthracycline-containing induction therapy followed by consolidation therapy as previously reported by the Japan Adult Leukemia Study Group (Yanada et al, Eur. J. Haematol. 2007). The relapse/refractory APL patients were received arsenic trioxide, tamibarotene, and/or gemtuzumab ozogamicin. Early death (death within 30 days from the start of antileukemic treatment), severe hemorrhagic events, and the improvement of coagulopathy were analyzed.

Results

The study population consisted of 97 males and 75 females, with a median age of 58 (range 10 - 88). TM-α was administration by 380 ± 52.5 U/kg/day and the duration of dosing was 7.0 ± 6.3 days. In most patients (81 %), TM-α was started on or before the start of antileukemic treatment. Within the first 30 days, 12 patients (7 %) had severe hemorrhage and there were 24 (14 %) early deaths. Six of those (3.5% of 172 patients) were due to hemorrhage. TM-α treatment rapidly and significantly improved coagulopathy regardless of concomitant ATRA treatment (Table 1).

Conclusions

Although anticoagulation therapy during APL chemotherapy may induce severe hemorrhagic complication, our findings suggest that supportive care with TM-α ameliorates coagulopathy and reduces the risk of hemorrhagic early deaths in patients with APL.

Disclosures:

Watanabe: Asahi Kasei Pharma Corporation: Employment. Honda:Asahi Kasei Pharma Corporation: Employment.