bluuebunny/crossref_metadata_2025_split huggingface dataset:
Minimal Residual Disease by Flow Cytometry in Children with Acute Lymphoblastic Leukemia. Single-Center Experience
Authors: Soria Marcela Eugenia , Gutierrez Marcela Ema , Gaillard Maria Isabel , Cores Maria , Gil Gimena , Moran Lorena Elizabeth , Rivas Pereira Maria Fernanda , Prada Silvina , Drelichman Guillermo , Feldman Leonardo , Aversa Luis Alberto 
Published: October 2019

Abstract
Abstract 4238

Introduction:
Minimal residual disease (MRD) allows detection of blasts in patients in morphologically complete remission. Detection of MRD in patients with ALL is an independent risk factor associated with lower event-free survival (EFS).

Objectives:
1) Evaluate MRD in bone marrowaspirateby flow cytometry at days 15 and 33 of induction in children with ALL. 2) Determine MRD association with hematologic features. 3) To correlate MRD association with overall survival (OS) and event-free survival (EFS).

Design and Statistical Analysis:
Descriptive, retrospective study. Univariate analysis was performed by T- test for continuous varibles and byχ2or Fisher tests for categorical variables. Survival was estimated with Kaplan Meier method. Regression multiple was used for multivariate analysis. p values < 0.05 were deemed as statistically significant. SPSS 15.0 program was used for the analyses.

Patients:
From January 2006 to December 2009, 84 patients with newly diagnosed ALL, aged 1 –18 years, were admitted at our institution. They were treated according to ALLIC / BFM / GATLA 2002 protocol. Immunophenotypic studies were performed by standard Four-color flow cytometry (FC) using FACSCalibur BD cytometer, MDR follow up panels were tailored depending on aberrant finding at diagnosis. 300.000–500.000 events were acquired using the CellQuest Pro and Paint -a-gate software for data analysis. MDR status has been defined as positive if at least 30–50 clustered events displaying leukemia –associated immunophenotypic characteristics (0,01%).

Results:
The mean age at diagnosis was 7.7 years (r: 2.1–18). Mean follow-up 33.9 months (r: 1–66). 38% were female. Immunophenotype: Pro B 5%, common B 87%, Pre B 2%, and T 6%. Good response to prednisone was achieved by 92% of patients. With regard to risk groups, the distribution was: 28% standard, 55% intermediate and 17% high. MRD at day 15 was evaluable in 66 patients (78%), being positive in 35 of them (45%). MRD at day 33 was evaluable in 75 patients (89%), being positive in 10 (12%).

Conclusion:
Positive MRD at day 15 was associated with: increased WBC count at diagnosis, high-risk group, higher relapse rate, lower EFS and OS. Otherwise, positive MRD at day 33 was associated: increased WBC count at diagnosis, T immunophenotype, poor response to prednisone, high risk group, lower EFS and OS.
Our data indicate that positive MRD at days 15 and 33 result an independent variable of poor prognosis in children with ALL.

Disclosures:
No relevant conflicts of interest to declare.
High Response Rate and Prolonged Survival in Patients with Primary Refractory Acute Myelogenous Leukemia Treated with Decitabine Salvage Therapy
Authors: Fernandez Ana , Shah Dhaval , Liu Delong , Ahmed Nasir , Lulaj Elizabeth , Seiter Karen 
Published: October 2019

Abstract
Abstract 4280
The prognosis for patients with AML who fail front line cytarabine/anthracycline-based chemotherapy is poor. Fewer than 10% of patients are expected to respond to second line chemotherapy and survival averages a few months. Decitabine is currently being used as a front line agent for patients with AML who are unfit for intensive chemotherapy generally because of advanced age. However in published studies the complete response rate in this population is only 25% (Cashen et al, JCO 2010). We recently treated 9 patients with primary refractory AML who failed cytarabine/anthracycline based chemotherapy with decitabine 20 mg/m2 × 5 days every 4–5 weeks. The median age was 59 years (range 40–83). There were 5 men and 4 women. Prior AHDs and comorbid conditions included MDS in 2 patients (both were treated with azacytidine), HIV in one patient, polycythemia vera in one patient, and breast ca treated with chemotherapy and radiation in one patient. Cytogenetics were normal in 2 patients, other intermediate in 3 patients and poor risk in 4 patients. Initial therapy of AML was cytarabine 3 gm/m2 × 5 and mitoxantrone 80 mg/m2 × 1 in 4 patients, 3+7 in 4 patients, and CPX-351 in 1 patient. 2 patients additionally had 2 courses of gemtuzumab ozogamicin with no response after front line chemotherapy and before decitabine. The median initial blast percentage prior to decitabine was 22% (range 7–73%), and after decitabine 5% (range 1–44%). 5 patients (56%) had a CR, 2 patients (22%) had a PR and 2 patients (22%) had progressive disease, including one who was extensively pretreated with azacytidine for prior MDS. The median number of decitabine cycles was 6 (range 1–12). Median time to progression was 11 months (range 1–20 months, with 2 patients remaining in CR at 10 and 14 months). 3 patients underwent allogeneic stem cell transplant after response to decitabine. One remains in CR 10 months post initiation of decitabine, and 6 months post bmt done in PR. One patient died of complications of GVHD 20 months post initiation of decitabine and 12 months post BMT, and one patient died of relapsed disease 18 months post initiation of decitabine and 10 months post BMT. The median survival is 12 months (range 10–20 months). We saw a high response rate to decitabine in patients with primary refractory AML. Although this could have occurred by chance due to the small sample size, we propose that decitabine might be more effective in patients who have undergone cytoreduction with intensive chemotherapy than in patients who have not received such therapy. Alternatively, lack of response to cytarabine/anthracycline therapy could select out a group of patients more likely to respond to decitabine.

Disclosures:
Off Label Use: Use of decitabine in AML. Seiter:Eisai: Speakers Bureau.
Recurrent Oncogenic Mutations in CCND3 in Aggressive Lymphomas
Authors: Schmitz Roland , Jhavar Sameer , Xiao Wenming , Liu Xuelu , Powell John , Wright George W. , Chan Wing C. , Jaffe Elaine S. , Gascoyne Randy D. , Campo Elias , Rosenwald Andreas , Ott German , Delabie Jan , Rimsza Lisa M. , Staudt Louis M. 
Published: October 2019

Abstract
Abstract 435
The recent development of high throughput mRNA sequencing technologies has allowed major advances in the characterization and quantification of transcriptomes in human cancer. These advances include gene mutation detection, gene fusion discovery, analysis of alternative splicing events, and unbiased gene expression profiling.
To comprehensively discover pathogenic sequence variants in lymphomas, we used Illumina technology to sequence mRNA of 206 lymphoma biopsies and cell lines, including 64 diffuse large B-cell lymphomas (DLBCL) of the activated B-cell-like (ABC) subtype, 71 DLBCL of the germinal center (GCB) subtype, and 41 Burkitt's lymphomas (BL). This effort uncovered a large number of mutations that were verified by Sanger sequencing.
Using this methodology, we discovered somatic mutations in CCND3, encoding Cyclin D3, in 38 % of BLs, 14 % of ABC DLBCLs, and 10 % of GCB DLBCLs. These mutations stabilized the Cyclin D3 protein by altering a phosphorylation motif at Thr283 that is required for proteasomal degradation. Knockdown of CCND3 by RNA interference confirmed the oncogenic addiction to this gene in the mutated cell lines. In parallel, an RNA interference screen revealed that the cell lines that are dependent on CCND3 are also reliant on cyclin-dependent kinase 6 (CDK6), which together with Cyclin D3 regulates the G1/S transition of the cell cycle. Ectopic expression of the mutant CCND3 isoforms accelerated lymphoma cell line proliferation thereby demonstrating the oncogenic potential of these mutations.
Remarkably, treatment of the CCND3/CDK6 addicted cell lines with a small molecule inhibitor of CDK 4/6 caused G1 arrest followed by apoptosis suggesting novel therapeutic strategies for these aggressive lymphomas.

Disclosures:
No relevant conflicts of interest to declare.
Mobilization of Autologous Peripheral Blood Stem Cells with Spectra Optia v5.0: A Single Center Experience with An Automatic Interface-Controlled Apheresis System
Authors: Eleutherakis-Papaiakovou Evangelos , Terpos Evangelos , Kastritis Efstathios , Christoulas Dimitrios , Migkou Magdalini , Dimopoulos Meletios Athanasios , Papadimitriou Christos 
Published: October 2019

Abstract
Abstract 4396
Available manual apheresis systems generally produce stem cell yields of consistently high quality that can be safely used for autologous transplantation. However, manual apheresis needs continuous interface monitoring/adjustement, suffers from interface instability in poor mobilizers, high collection variability, high platelet loss and failure to electronically document parameters. A new advanced apheresis system, Spectra Optia v5.0, featuring optical sensors, which provide real time automatic interface and product line control, was designed to override these disadvantages.
In our study, we evaluated data of 10 stem cell leukaphereses performed in 8 patients with various malignancies using Spectra Optia during 2011 to test its feasibility and effectiveness. We compared our data with those obtained from 225 patients that had undergone a stem cell collection for autologous transplantation in our Department between 2004 and 2011, using the COBE Spectra machine. The use and function of automatic interface control of Spectra were satisfactory. Due to the application of lower inlet volumes/min, as compared to corresponding volumes with the COBE Spectra machine, our apheresis with Spectra Optia usually took a longer time (median 447 min versus 317 min, p<0.005). Regarding other collection parameters, such as the percentage of CD34+ cells in the final leukapheresis product, total yield of CD34+ cells, and product volume data were comparable for both devices (0.74% versus 0.97% CD34+, p=0.103; 6.85 versus 7.1 × 106 CD34+/kg, p=0.752; and 403.5 versus 353 ml graft volume, p=0.094, respectively). Time to engraftment was also comparable for both apheresis devices. More specifically, time interval to neutrophil counts >500/μl, neutrophil counts >1500/μl did not significantly differ (10 days versus 9 days, p=0.386 and 11 days versus 10 days, p=0.229, respectively). However a delay in platelet recovery with Optia device need to be confirmed with additional data from apheresis procedures (median time to platelet counts >25000/μl was 12 days versus 11 days for COBE Spectra, respectively; p=0.037). Platelet loss with Optia was less than with COBE Spectra (1278 versus 2415 × 103/μl, p=0.014). No significant differences were observed for product hematocrit between Optia and COBE Spectra (5.7% versus 6.6%, p=0.392).
In conclusion, the automatic Spectra Optia aphereses were associated with similar and equally variable stem cell collections as aphereses with COBE Spectra. Further data are needed to clarify the potential benefit of lower platelet loss using Optia. We continue to use this procedure in our center and updated results will be presented in the meeting.

Disclosures:
No relevant conflicts of interest to declare.
The Outcome of Autologous Stem Cell Transplantation and the Risk of Relapse in Myeloma and Lymphoma Patients Mobilized Using Plerixafor Compared to Standard Mobilization Regimen
Authors: Zielinska Patrycja , Krawczyk-Kulis Malgorzata , Markiewicz Miroslaw , Koclega Anna , Liwoch Robert , Dzierzak-Mietla Monika , Bialas Krzysztof , Kyrcz-Krzemien Slawomira 
Published: October 2019

Abstract
Abstract 4399
Autologous stem cell transplantation (ASCT) is a method of choice in many hematological malignancies enabling the patients (pts) to achieve remission or to prolong life. Commonly used mobilizing regimens include high dose chemotherapy followed by G-CSF administration. Unfortunately up to 35% pts fail to mobilize the successful number of stem cells (SC).
16 pts aged 48 (23–69) were mobilized with plerixafor in our clinic over one-year period. All of the them failed to mobilize to collect satisfactory number of SC after standard mobilizing regimen. The protocol included G-CSF (10μg/kg) administered subcutaneously for 4 days, followed by plerixafor administration (240μg/kg s.c.). Only two pts failed to collect satisfactory number of SC after plerixafor. 14/16 collected the median of 2.24×106/kg (0.8–15,8), all of them underwent the ASCT. The outcome of ASCT in this group was compared to the posttransplant period in pts mobilized with standard mobilizing regimen (control group; CG).
The amount of CD34+cells x106/kg was significantly higher in the CG (2.24 vs. 4.4, p<.05). The proportion of CD34+ cells among total nucleated cells was significantly lower in the plerixafor group compared to the CG (0.22 vs 1.72, p<.004). The high nucleated cell count translated into high volume of SC product (median 1650mL; 800–2800mL). The volume of frozen SC product was significantly higher in the plerixafor group compared to the CG (1650 vs. 800mL, p<.00). The length of hospital stay (median: 26 days in both groups), incidence of serious infections (41% in plerixafor group vs. 40% in the CG), use of i.v. antibiotics (median: 15 vs. 16 days), number of RBC and platelet (PLT) transfusions (median: 2 vs. 3 units for both RBC and PLT), time to ANC engraftment (ANC>0.5G/l) −10 vs. 14 days, and to PLT engraftment (PLT>20G/l) – 11vs. 14 days, were not different significantly. 43% pts relapsed in plerixafor group with a median follow-up of 14 months and 37% pts in the CG with a median follow up of 11 months (p>.05).SC mobilization with plerixafor and G-CSF provides solution for majority of pts requiring ASCT and failing mobilization with G-CSF in combination with chemotherapy. However, due to high leukocytosis, this protocol may require modification of SC collection and freezing procedures in order to avoid large volumes. In general, pts mobilized with plerixafor had similar post transplant outcome as pts who were mobilized using standard regimen.

Disclosures:
No relevant conflicts of interest to declare.
Remobilization with High Dose Methotrexate and Cytarabine in Patients with Non-Hodgkin Lymphoma and Multiple Myeloma Previously Failing Mobilization with Chemotherapy and Cytokine Treatment
Authors: Park Seong Joon , Yoon Dok Hyun , Kim Shin , Suh Cheolwon 
Published: October 2019

Abstract
Abstract 4402

Background
High-dose chemotherapy supported by autologous stem cell transplantation is a widely used therapeutic modality especially in patients with non-Hodgkin lymphomas (NHL) and multiple myeloma (MM). However, mobilization failure rates following first attempt are estimated to be between 5% and 30% and remobilization is required in those patients. Although plerixafor, a selective CXCR4 antagonist, is a promising agent for these purpose, mobilization with chemotherapeutic agents would be a hands-on option.°° Therefore we investigated the feasibility of high dose methotrexate and cytarabine with G-CSF as a remobilization regimen in those who failed a prior mobilization and collection with chemotherapy and G-CSF.

Patients and methods
Mobilization failure was defined as a collection of less than 5 × 106 CD34+ cells after 3–5 apheresis procedures. Methotrexate (3,500 mg/m2 in a 120 min infusion) on day 1 and cytarabine (3,000 mg/m2 infusion for 120 min) on day 4 and day 5 were followed by G-CSF. Supportive care and leucovorine rescue were done as standard protocol. Peripheral blood (PB) hematopoietic progenitor cells (HPCs) were counted with the Sysmex SE9000. PB progenitor cell harvest was initiated when HPC levels reached at least 5 per mm3.

Results
Total 8 patients (6 NHL and 2 MM, median age: 55 years) who have failed in previous mobilization with conventional chemotherapy and G-CSF were tried again with high dose methotrexate, cytarabine and G-CSF. (table 1) Successful collection of CD34+ cells (&gt; 5 × 106/kg) was achieved in six patients (75%). The total yield of CD34+ cells per kg of body weight harvested by 3 to 5 apheresis procedures was 6.28 × 106/kg (median; range, 1.53– 10.09 × 106/kg). (table 2)

Conclusion
High dose-MTX and cytarabine plus G-CSF may be a feasible chemotherapeutic regimen for stem cell mobilization in NHL and MM patients who have failed in previous mobilization with other type of chemotherapy and G-CSF.

Disclosures:
No relevant conflicts of interest to declare.
Ursolic Acid Induced Apoptosis Involving Upregulation of Expression of PTEN Gene and Inactivation of PI3K/Akt Pathway in K562 Cells
Authors: Wu Bin , Hu Rong , Wang Huihan , Wang Hongtao , Zhang Rong , Yang Wei , Liu Zhuogang 
Published: October 2019

Abstract
Abstract 4453

Object
Ursolic acid(UA), a pentacyclic triterpenoid derived from many kinds of medicinal plants, exhibits potent anticancer activity in many kinds of cancer cells. However, the anticancer mechanism of UA is not clearly understood.

Method
The toxic action of UA in K562 cells was assessed using CCK-8. K562 cells trested by UA were incubated with FITC-conjugated Annexin V and counterstained with PI in order to allow exclusion of necrotic cells, then were subsequently analyzed using a flow cytometer. Pro-caspase-3, cleaved caspase-3, Procaspase-9, cleaved caspase-9, Akt and Akt phosphorylation were analyzed by Western Blotting. The expression of PTEN gene is analyzed by Quantitative real-time RT-PCR.

Result
Here we showed that UA could cause growth inhibition, and induce apoptosis in human chronic myelogenous leukemia cell line K562 cells. The expression of PTEN gene is rejected in many kinds of cancers including leukemia, then the PI3K/Akt pathway is actived. So the PTEN gene and PI3K/Akt pathway become the central targer for cancer therapy. UA treatment can up-regulate the expression of PTEN gene, inhibite the activity of Akt kinase, change the mitochondrial transmembrane potential, then induce the reduce of cytochrome C and the active of caspase family.

Conclusion
These results suggest that UA might exhibit its strong antitumor effects via the up-regulation of PTEN gene and the inhibited the activity of PI3K/Akt pathway.

Disclosures:
No relevant conflicts of interest to declare.
Allogeneic Hematopoietic Stem Cell Transplantation Using Reduced-Intensity Conditioning in the Patients with Acute Leukemia
Authors: Kim Hyun-Kyung , Mun Yeung-Chul , Yoo Eun-Sun , Lee Kyoung Eun , Nam Eunmi , Lee Soon Nam , Seong Chu-Myong 
Published: October 2019

Abstract
Abstract 4508

Aim:
Allogeneic hematopoietic stem-cell transplantation (HSCT) following myeloablative regimens has been used to treat patients with acute leukemia. Reduced-intensity conditioning (RIC) has been developed for older patients which are associated with a high risk of transplant-related mortality (TRM). We investigated the outcomes of allogeneic HSCT with RIC in the patients of acute leukemia.

Method:
Among the patients with acute leukemia who have received allogeneic HSCT in Ewha Womans University Hospital between 1998 and 2010, outcomes of the patients who received HSCT with RIC were analyzed retrospectively.

Result:
Twenty-two patients were enrolled in this study including 8 patients of acute lymphoblastic leukemia (ALL) in first complete remission (CR), 14 patients of acute myeloid leukemia in first CR (n=9), second CR (n=2), and not in CR (n =3). Seventeen patients with hematopoietic cell transplant comorbidity index score 0 were included in this analysis. Twelve patients received fludarabine (150mg/m2) with melphalan (140mg/m2), and 10 patients received fludarabine (150mg/m2) with busulfan (6.4mg/m2) as conditioning regimen, and followed by transplantation from matched sibling (n=16), unrelated donors (n=5), or unrelated cord blood (n=1). All patients received graft-versus-host disease (GVHD) prophylaxis with methotrexate and cyclosporine (n=18) or FK506 (n=3) except one patient who received cyclosporine alone. Cumulative incidence of grade 2–4 acute and chronic GVHD were 18.2% and 45.5%. Donor chimerism in bone marrow at 30 day and in blood at 100 day were median 99.0% and 97.0% after HSCT. Rates of relapse was 22.7% after median 7.5 months of follow-up. Non-relapse mortality at 100 day and 1 year were 13.6% and 27.3%. Disease-free survival and overall survival were median 33.0 month and 42.7 month by Kaplan-Meier analysis.

Conclusion:
These results suggest that allogeneic HSCT using RIC for treatment of acute leukemia may be a potential therapeutic approach considering less toxic and acceptable treatment outcomes even in young adult ALL without comorbidity which has not been previously considered as candidate of allogeneic HSCT using RIC.

Disclosures:
No relevant conflicts of interest to declare.
The Impact of Co-Morbidities on the Outcome of Autologous Stem Cell Transplantation for Multiple Myeloma
Authors: Saad Ayman A , Sala Marc , Naik Niravkumar A. , Hari Parameswaran 
Published: October 2019

Abstract
Abstract 4511

Introduction:
Autologous stem cell transplantation (ASCT) is used as an upfront and salvage therapy for multiple myeloma (MM). We assessed the prognostic value of pre-transplant co-morbidity on the transplant outcome.

Method:
We analyzed the data of 162 patients with MM who underwent single ASCT between 2004 and 2009 in our center. Pre-transplant co-morbidity was scored according to the hematopoietic cell transplant co-morbidity index (HCT-CI). Data were tabulated and statistical difference was calculated using t-test and Chi square test. Correlation coefficiency test was used to correlate HCI-CI with data of continuous variables. Probabilities of overall survival were analyzed using Kaplan-Meier analysis.

Result:
The median follow up after transplant was 23 months (range: 0.7–78). The median age at transplant was 60 years (range: 34–77), males were 64%. A total of 130 patients (80%) had transplant at ≤ 12 months from diagnosis. The proportion of IgG, IgA and free light chain subtypes was 54, 24 and 17% respectively. Durie Salmon stage at diagnosis was stage I (8%), II (12%), III (60%) respectively. The median HCT-CI was 2 (range: 0–8). HCT-CI was 0, 1–2, >2 in 27%, 32%, and 41% respectively. There was no significant correlation between HCT-CI and time to engraft or length of stay. Patients with HCT-CI of 0–2 (group 1; n= 95) and >2 (group 2; n=67) had a re-admission rate (within 30 days) of 13.7% and 21% respectively (p=0.23). Group 1 had non-relapse mortality (NRM) of 0% at 18 months, while group 2 had NRM of 2% and 2.5% at 6 months and 18 months respectively (p= 0.02). There was no significant difference in the OS; not reached and 60 months in group 1 and 2 respectively (p=0.3).

Conclusion:
These data show that high pre-transplant HCT-CI is associated with higher NRM following ASCT for multiple myeloma. This finding needs to be validated in a large cohort of patients.

Disclosures:
No relevant conflicts of interest to declare.