bluuebunny/crossref_metadata_2025 huggingface dataset:
Conditioning with Treosulfan and Fludarabine for Patients with Refractory or Relapsed Non-Hodgkin Lymphoma (NHL),
Authors: Schmitt Michael , Trenschel Rudolf , Sayer Herbert G. , Schneider Catarina , Glass Aenne , Hilgendorf Inken , Treschl Anne , Junghanss Christian , Borchert Kersten , Koenigsmann Michael , Casper Jochen , Beelen Dietrich , Freund Mathias , Kahl Christoph
Published: October 2019

Abstract 4106

Background:

Treatment of refractory or relapsed non-Hodgkin lymphoma (NHL) remains challenging. Here, 88 patients with refractory or relapsed NHL received treosulfan and fludarabine as a reduced intensity conditioning for allogeneic hematopoietic stem cell transplantation (allo-HSCT).

Material and Methods:

73/88 intensely pre-treated patients experienced a relapse (R) with 18/88 early relapses (ER < 6 months from the last chemotherapy). At the time of allo-HSCT, 26 patients were in complete (CR) and 43 in partial remission (PR), twelve patients had progressive disease (PD) and seven stable disease (SD). 47 patients received an autologous graft followed by allo-HSCT.

Results:

After allo-HSCT, 69 of 88 patients were in CR, seven patients in PR, resulting in an overall response rate of 86.4% (76/88). Thirty-three patients achieved a CR from PR as well as six patients from PD and five from SD. 43/88 (49 %) patients were alive at the end of follow-up. Patients undergoing directly allo-HSCT without preceding auto-HSCT showed a better disease-free survival (DFS, p =.038) with a trend (p =.077) for better overall survival (OS). Patients with ER showed an OS of.35 ±.12 after three and seven years. Chronic graft-versus-host disease (cGvHD) had a positive impact on both OS and DFS (for limited cGVHD versus no cGvHD p =.002 and.004, respectively).

Conclusion:

Allogeneic stem cell transplantation following conditioning with treosulfan and fludarabine constitutes a good therapeutical option for patients with refractory or relapsed NHL and should be considered early in the course of salvage treatment.

Disclosures:

Schmitt: Medac: Participation in Conferences. Sayer:Medac: Travel grant to ASH 2009. Koenigsmann:Medac: Research Funding. Casper:Medac: Participation in Conferences. Beelen:Medac: Participation in Conferences. Freund:Medac: Honoraria, Research Funding.
NK Cell Activity Influences Long Term Outcome of Pediatric Leucemias After T Cell Depleted Stem Cell Transplantation,
Authors: Lang Peter , Pfeiffer Matthias , Teltschik Heiko-Manuel , Schlegel Paul G. , Feuchtinger Tobias F. , Klingebiel Thomas , Beck James F. , Handgretinger Rupert
Published: October 2019

Abstract 4141

T cell depletion with magnetic microbeads can effectively reduce GvHD rates after stem cell transplantation from both mismatched related donors as well as from matched or partially matched unrelated donors. However, T cell recovery is markedly delayed after this procedure and T cell mediated antileukemic effects may be reduced. Thus, we focused on the rapidly regenerating donor derived NK cell system and addressed the question, whether its functional activity would influence the probability of relapse in a long term analysis. Temporal development of NK cell activity was monitored in 47 pediatric patients with leukemias (ALL, AML, CML, JMML) and myelodysplastic syndromes after transplantation of T cell depleted stem cells from matched unrelated (n=18) and mismatched related (haploidentical, n=29) donors with a median follow up of 7.4 years (2.1–12). 38 patients had CR1-3, 9 patients had active disease at time of transplantation. EFS and relapse rate at 5 years for the entire group were 36% and 40%, respectively (EFS and relapse rate for ALL patients in CR1-3: 50% and 36%; EFS and relapse rate for AML/MDS patients: 22% and 30%). CD34+ selection with magnetic microbeads resulted in 8×103/kg residual T cells. No posttransplant immune suppression was given. 89% of the patients had no GvHD, 9% had GvHD grade I and only 2 patients had GvHD grade II or III. NK cells recovered rapidly after transplantation (300 CD56+/μL at day 30, median), whereas T cell recovery was delayed (median: 12 CD3+/μL at day 90). NK activity was measured as specific lysis of K562 targets several times after transplantation (mean: 3 assays per patient). Four temporal patterns of lytic activity could be differentiated: consistently low, consistently high, decreasing and increasing activity. Patients with consistently high or increasing activity had significantly lower relapse probability than patients with consistently low or decreasing levels (0.18 vs 0.73 at 5 years, p<0.05). The subgroup of patients with ALL showed similar results (0.75 vs 0.14 at 5 years, p<0.05). Speed of T cell recovery had no influence. These data suggest that both achieving and maintaining a high level of NK activity may contribute to prevent relapse. Thus, this model comprises direct functional data in the form of NK cell activation levels which are likely to be influenced by the presence of different cytokines in each patient. Our observations may have some clinical implications: immunomagnetic depletion of T cells prevents GvHD and can be performed in pediatric leukemias in remission without excessive increase in relapse rates. High levels of NK activity seem to be of importance. Since NK activity could be markedly increased by in vitro stimulation with Interleukin 2 (IL-2), in vivo administration should be considered.

Disclosures:

No relevant conflicts of interest to declare.
The Favorable Results of Allo-HSCT in TKI-Resistant CML,
Authors: Markiewicz Miroslaw , Dzierzak-Mietla Monika , Wisniewska-Piaty Katarzyna , Frankiewicz Andrzej , Koclega Anna , Zielinska Patrycja , Wieczorkiewicz-Kabut Agata , Sobczyk-Kruszelnicka Malgorzata , Bialas Krzysztof , Kyrcz-Krzemien Slawomira
Published: October 2019

Abstract 4145

Introduction:

Tyrosine kinase inhibitors (TKI) have revolutionized the treatment of chronic myeloid leukemia (CML). However, for patients who fail TKI or progress to advanced phase disease, allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains the only therapeutic option. The rationale of this study was to evaluate results of allo-HSCT in CML patients who have failed TKI treatment. Patients and Methods: 48 CML pts aged 33 (19–52) years who failed previous treatment with TKI (imatinib-37 pts, imatinib+dasatinib-5, imatinib+dasatinib+nilotinib- 3, dasatinib-2, imatinib+nilotinib-1) received allo-HSCT from HLA-matched siblings (15) or from 10/10 (21) and 9/10 (12) HLA-antigens-Matched Unrelated Donors (MUD) in Hematology and BMT Center in Katowice, Poland, from 10.2002 to 03.2011. The myeloablative preparative regimen consisted of treosulfan 3×14 g/m2 plus fludarabine 5×30 mg/m2 (30 pts) or busulfan 4×4 mg/kg plus cyclophosphamide 4×30 mg/kg (18 pts). Standard GVHD prophylaxis consisted of cyclosporine-A, methotrexate and pre-transplant ATG or thymoglobulin in allo-HSCT from MUD. Source of cells was bone marrow (27 pts) or peripheral blood (21 pts) with median 2.6 or 6.7 x10(6)CD34+cells/kg, respectively. Results: All pts engrafted. 100% donor chimerism has been achieved in 40 (83.3%) pts, 97–99% in 4 pts, progressing mixed chimerism was observed in 4 pts. The 5-year estimated overall survival rate was 79%. 9 pts died 9 (1–21) months following allo-HSCT due to infection (5), GVHD (3) or relapse (1). Bio-molecular relapse or progressing mixed chimerism was observed in 6 pts and was treated with 3 to 5 donor lymphocyte infusions (3 pts), post-transplant imatinib (3 pts) or nilotinib (1 pt). Acute GVHD grade I, II, III and IV was observed in 15, 13, 2 and 1 pt (serious aGVHD grade III-IV in 3 pts (6.25%) only); limited and extensive chronic GVHD in 17 (35.4%) and 9 (18.75%) pts, respectively. Other complications in survivors included CMV reactivation (12), hemorrhagic cystitis (3), H. zoster (2), P. jiroveci (1) and cataract requiring surgery (1). 39 pts (81.25%) are alive 44 (3–92) months post-transplant. Conclusions: Allo-HSCT with myeloablative treo/flu or bu/cy conditioning is a feasible and effective curative therapy in TKI-resistant CML. The alternative therapy with allo-HSCT may overcome TKI resistance, providing long-term remission or cure from CML in patients who failed TKI treatment.

Disclosures:

No relevant conflicts of interest to declare.
Development of the Functional Assessment of Cancer Therapy-Multiple Myeloma (FACT-MM) Scale and Validation in the Eastern Cooperative Oncology Group Trial E1A05,
Authors: Weiss Matthias , Jacobus Susanna , Wagner Lynne I , Cella David , Katz Michael S , Rajkumar Vincent , Fonseca Rafael
Published: October 2019

Abstract 4184

Introduction:

FACT-MM was developed with the aim to create a disease-specific, patient-reported outcomes (PRO) measure as part of the FACT measurement system to assess multiple myeloma (MM)-related symptoms. Literature review identified 52 MM specific symptoms and concerns. 13 MM expert clinicians rated these 52 items on relevance to health-related quality of life (HRQL) for MM patients and added 11 items for comprehensive PRO assessment in MM. These 63 candidate items were rated by 13 MM patients recruited through the International Myeloma Foundation website. Disease-related symptoms and concerns were described and provided by patients through free-text comments. Information from both the MM expert clinician and MM patient surveys including free-text items was analyzed and 14 highest ranked items were selected for the FACT-MM. The FACT-MM subscale (score 0–56), the FACT-G physical and functional well-being subscales (score 0–28), and the FACT-Neurotoxicity (FACT-Ntx) subscale (score 0–44) was administered to 48 E1A05 participants to assess disease and treatment-related symptoms of patients with newly diagnosed MM receiving 8 cycles of VRd versus Vd. Methods: In this study, the first three of seven sequential assessments were evaluated: baseline, cycle 5 and end of treatment (after cycle 8 or early diconstinuation). Instruments were scored as per the FACT scoring guidelines. Descriptive statistics were provided for each timepoint and changes in scores from baseline. The Wilcoxon rank sum test was used to assess differences in scores by ISS stage and ECOG PS. Pearson correlation coefficients were obtained between the scores. Cronbach's alpha was used to evaluate internal consistency of the FACT-MM. Results: At baseline and end of treatment, 46 and 41 patients, respectively, completed assessments. The FACT-MM demonstrated good to very good internal consistency (Cronbach alpha 0.79 – 0.89). The FACT-MM subscale was significantly correlated with the FACT physical and functional well-being subscales at baseline (r = 0.72), cycle 5 (r = 0.64; r = 0.49) and end of treatment (r = 0.72; r = 0.66). All scores declined modestly by cycle 5 and end of treatment from baseline. While sample size was limited, there appeared association with ECOG PS with higher scores for patients with PS 0 status. Conclusions: The 14-item FACT-MM scale is feasible for use to measure myeloma-related symptoms and has demonstrated acceptable psychometric properties based on findings from E1A05, an ECOG myeloma trial.

Disclosures:

Cella: Novartis: Research Funding. Fonseca:Consulting:Genzyme, Medtronic, BMS, Amgen, Otsuka, Celgene, Intellikine, Lilly Research Support: Cylene, Onyz, Celgene: Consultancy, Research Funding.
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 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 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 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 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 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.
Quality of Cord-Blood Units and Hematological Recovery After Intra Bone CBT Are Not Influenced by the Bank of Origin and by the Time of Cryopreservation
Authors: Kurita Naoki , Frassoni Francesco , Sacchi Nicoletta , Cossu Claudia , Serio Alberto , Cilloni Daniela , Podestà Marina
Published: October 2019

Abstract 4400

BACKGROUND:

Cord-blood transplantation (CBT) is an effective option for both malignant and non-malignant hematological diseases and its increasing number in the last years has enlarged the frequency of the use of cord-blood units (CBUs) trough international borders in order to maximize the opportunity of identifying optimal CB unit. Since there is no global standard regarding cord-blood quality control, the quality of CBUs can be affected by the bank of origin. In addition, few information are disposable on the cord blood engraftment capacity after long-term cryopreservation. In this study, we evaluated whether the origin and the age of CBUs may influence the quality of CBUs and post-transplant hematological recovery.

METHODS:

CBTs with single unit were performed at San Martino Hospital in Genoa for hematological malignancies using an intra-bone cord blood transplant technique. Just before infusion, a small sample of the CBU was obtained and post-thawing number of total nucleated cells (TNCs), CD34 positive cells, and colony-forming cells (CFCs) were characterized. Median pre-freezing number of TNCs and CD34-positive cells were 2.32 (1.15 – 4.52) × 109 and 9.3 (2.4 – 27.24) × 106, respectively. Recovery of neutrophil and platelet were compared according to the origin and the age of each CBU.

RESULTS:

95 CBUs were tested. The bank of origin of CBUs was Europe (n = 45), the United States (n = 40), and other regions (n = 10). Median age of CBUs was 4.9 (0.4 – 11.8) years; CBUs originated from European bank were significantly older than CBUs from other regions (5.9 years vs 3.8 years, p value of unpaired t-test < 0.001). Median post-thawing number of TNCs, CD34 positive cells, and CFCs were 2.3 (0.74 – 4.5) × 109 cells, 1.1 (0.23 – 2.7) × 107 cells, and 1.8 (0.0 – 12) × 106 cells, respectively. These values did not differ considering older (> 4.9 years) CBUs or younger CBUs (< 4.9 years) or the bank of origin. Rate and median time of neutrophil and platelet engraftment was as follows: younger CBUs: 24 and 34 days, older CBUs: 25 and 35 days, respectively (Figure 1,3). Hematological recovery was not influenced by the bank origin of CBUs as well, namely, rate and time of neutrophil and platelet engraftment was, 22 and 34.5 days (Europe), 22 and 35 days (the United States), 24 and 42 days (the other region), respectively (Figure 2,4).

CONCLUSION:

Although in our experience time of cryopreservation of CBUs was significantly dependent from bank of origin, the origin and the age of CBUs have little impact on post-thawing values of TNCs, CD34 positive cells, and CFCs. In addition, hematological recovery after transplant was not influenced by the origin or by the age of CBU. These data suggest that quality control of CBU is working well regardless of the bank origin.

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 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 (> 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 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 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 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.
The Outcome of Allogeneic Stem Cell Transplantation in Chronic Lymphocytic Leukaemia: A Single Centre 10-Year-Experience
Authors: Zielinska Patrycja , Krawczyk-Kulis Malgorzata , Markiewicz Miroslaw , Dzierzak-Mietla Monika , Koclega Anna , Liwoch Robert , Kopinska Anna , Kyrcz-Krzemien Slawomira
Published: October 2019

Abstract 4515

Chronic lymphocytic leukemia (CLL) is an incurable disease when treated with standard chemotherapy. The only possibility to provide cure is allogeneic stem cell transplantation (allo-SCT). CLL patients aged less than 55 account for about 15% of patients and these cases allo-SCT should be taken into consideration. The indications for allo-SCT are as follows: del17p, resistance to chemoimmunotherapy, Richter’s syndrome or recurrent disease. A retrospective analysis of allo-SCT in 18 patients (10 males, 8 females) with CLL transplanted in years 2000–2010 was performed. The aim of the study was to assess of long term follow-up outcome of allo-SCT in CLL patients. The median age at diagnosis was 41ys (range: 35–51). The sibling donor was available in 16 cases (2 pts were mismatched), unrelated donors were in 2 cases (1 mismatched). Most of the pts (16 out of 18) were MRD positive when allotransplanted. Median lymphocytosis preceeding allo-SCT was 5.9G/l. Peripheral blood was the source of stem cells in 9 cases (50%), and bone marrow in the remaining 9 cases, 2 pts were transplanted with stem cells from bone marrow and peripheral blood. 4 pts (22%) underwent the allograft procedure twice or more. Reduced intensity conditioning with alemtuzumab was performed in 9 pts (50%), myeloablative regimen in 4 cases and RIC with rituximab in one case.The median number of CD34+cellsx10^6/kg was 4.1 (range: 0.86–9.64). All but one patient engrafted (this pt was transplanted again successfully in one year time). Acute graft-versus host disease (GvHD) was noted in 46% of pts (only in 2 pts grade IV). Extensive GvHD was observed only in 2 pts. Donor lymphocyte infusion (DLI) was performed in 8 pts (44%). With a median follow-up of 73 months (range: 9–89) for surviving patients, the five-year Kaplan-Meier of overall survival (OS) and progression free survival (PFS) was 55,5% and 34%, respectively. At five years, the cumulative probability of non-relapse mortality was 15%. Allogeneic stem cell transplantation remains the effective treatment in CLL for selected group of patients.

Disclosures:

No relevant conflicts of interest to declare.
Effect Of Plerixafor Mobilization On Immune Reconstitution Post High Dose Chemotherapy and Autologous Stem Cell Transplantation
Authors: Solh Melhem M. , Smith Tori , Khaled Yasser , Copik Alicja
Published: October 2019

Introduction

Plerixafor is a reversible CXCR4 antagonist that has been approved by the food and drug administration for autologous hematopoietic stem cell mobilization in patients with multiple myeloma and non-Hodgkin’s lymphoma. Patients mobilized with Plerixafor were shown to have a higher proportion of primitive stem cells (CD34+/CD133+/CD38-), CD4+ T cells and Natural killer cells (CD3-/CD16+/CD56+) in the graft composition when compared to patients mobilized with chemotherapy plus G-CSF alone .We investigated the effect of Plerixafor on immune reconstitution at thirty and sixty days post autologous stem cell transplantation.

Methods

Patients eligible for autologous stem cell transplantation were enrolled on a single arm prospective immune reconstitution trial. A complete blood count, differential and lymphocyte flow cytometry panel (T cell, NK cell and B cell markers) was checked on Days 30 and 60 post autologous transplantation. Stem cell mobilization was carried per our institutional standards. All patients received subcutaneous G-CSF at a dose of 10 µg per kilogram body weight for four consecutive days. On Day4, patient with a peripheral CD34 count of ≤20/µl received plerixafor 0.24mg/kg. Collection was started on day 5 and continued till collection goal was reached or patient failed to get the minimal cell dose after 4 consecutive days of aphaeresis.

Results

49 patients were enrolled during the period from September 2010 till May 2012. Median age at time of transplantation was 54 years (range 21; 72 years). 35 patients had multiple myeloma and 14 had non-Hodgkin’s lymphoma. 16 patients received GCSF alone (group A) and 33 had plerixafor plus GCSF (group B) for mobilization. All patients achieved the minimum target of CD34 collection. The mean number of collection days was 1.9 and 1.4 days (p=0.05) with a total collection dose of 7.76 and 7.61 CD34 x106/Kg for groups A and B respectively. The percentage proportion of CD34 in the aphaeresis product was 0.73% and 0.75% (p=0.9) for group A and B. Total infused CD34 dose was similar in both groups (4.88 and 4.56 CD34x106/kg) with time to engraftment of 11.68 vs 11.69 days for neutrophils and 20.62 vs 21.39 for platelets in groups A and B respectively. There was no difference between day 30 absolute lymphocyte count (1.09 vs 1.44 x103/mm3 p=0.18); Absolute NK cell (0.31 vs 0.35 x103/µl; p=0.51); absolute T cell count (0.71 vs 0.96 x103/µl p=0.33) and absolute neutrophil count (2.98 vs 2.63 x103/mm3 p=0.37). The cell count recovery was also not significantly different when analyzed per disease (myeloma or non-Hodgkin’s lymphoma) and at day 60.

Conclusion

Our study shows that patient mobilized with plerixafor and G-CSF have similar immune reconstitution at 30 and 60 days post autologous transplantation compared to patients mobilized with G-CSF alone.

Disclosures:

No relevant conflicts of interest to declare.
Effect Of Allogeneic Blood Transfusion On Peripheral Blood Τregs and Cytokine Levels Of Patients That Underwent Joint Replacement Surgery
Authors: Spadidea Panagiota , Rodi Maria , Chatziantoniou Alexandros , Panagiotopoulos Elias , Mouzaki Athanasia
Published: October 2019

Purpose/Objective

Clinical and experimental studies have established that allogeneic blood transfusion (ABT) can cause immunosuppression. To identify immune parameters that contribute to this effect, we determined the effect of ABT on peripheral blood (PB) cytokine profiles and Treg numbers and function in a cohort of patients with no underlying pathologies.

Materials and Methods

Heparinized PB samples were collected from 46 patients (7M/39F, 28-88 yo) that underwent joint replacement surgery. The samples were collected immediately before surgery (BS), and after surgery (AS) on days 0, 7, 1 month, and 3 months to 1 year. Thirty six patients received ABT and 10 did not. PBMC were isolated, and the numbers and % of CD4+CD25+Foxp3+ Tregs and CD4+CD25high/+CD127low/- Tregs were determined by FACS. Tregs and T effectors (Teff) were isolated from patients on days 0-7 and Treg functional assays were performed by culturing Tregs with PHA-stimulated Teff at different ratios for 72h with CFSE, and analyzed by FACS for proliferation. Cytokine levels were determined in plasma by a cytometric bead array assay for IL-2, IL-4, IL-5, IL-6, IL-10, TNF-á, IFN-ã, and ELISA for TNF-á, TNF-RI(p55/p60) and II(p75/p80), TGF-â1 and â2.

Results

Both, CD4+CD25+Foxp3+ and CD4+CD25high/+CD127low/- Treg populations increased significantly on d0 AS and decreased on d7 AS to below BS levels, to achieve homeostasis >3 months in transfused patients. In contrast, Treg levels remained the same between BS and AS in non-transfused patients.

Functional assays showed that Tregs were functional post-ABT and could suppress Teff proliferation efficiently. In culture, isolated Tregs secreted TGF-â1.

All cytokines and TNF-RI and II plasma levels increased on d0 AS (IL-6, TNF-RI and II significantly), in transfused patients immediately AS, and decreased by d7 AS. In contrast, TGF-â1 levels decreased on d0 AS and increased by d7. No differences in cytokine or receptor levels were observed in non-transfused patients AS.

Conclusion

ABT induces the immediate production of plasma cytokines, especially pro-inflammatory IL-6 and TNF-RI and II that decreases by d7. In parallel, ABT induces the proliferation of Tregs. These Tregs are functional, and secrete TGF-â1 that reaches BS plasma levels by d7. The Tregs seem to be Th3 inducible Tregs. Homeostasis is reached by 3 months post-ABT.

Disclosures:

No relevant conflicts of interest to declare.
OSU-T315, An Integrin-Linked Kinase (ILK) Inhibitor, Induces Apoptosis By Targeting B Cell Receptor and CD49d Mediated AKT/ERK Activation In Chronic Lymphocytic Leukemia Cells
Authors: Liu Ta-Ming , Ling YongHua , Phelps Mitch A. , Woyach Jennifer A. , Hertlein Erin K , Towns William H. , MacMurray Jessica , Salunke Santosh B. , Chen Ching-Shih , Byrd John C. , Johnson Amy J.
Published: October 2019

Aberrant regulation of endogenous survival pathways plays a major role in pathogenesis of chronic lymphocytic leukemia (CLL). Signaling via conjugation of surface receptors within the tumor environmental niche is believed to activate survival and proliferation pathway in CLL. PI3K/AKT appears to be the pivotal axis to support CLL growth. Current studies identify Integrin-linked kinase (ILK) as a Phosphoinositide-dependent kinase-2 (PDK2), by which it mediates AKT activation and concomitant with tumorigenesis in many solid tumors. As a result of our search for novel therapeutic targets, we first report the potent inhibitor, OSU-T315, modeled from the scaffold that docks into AKT-binding site of ILK in therapeutic of hematological malignancy. OSU-T315 triggers apoptosis in CLL by targeting both intrinsic and extrinsic AKT and ERK activation (Figure 1A). OSU-T315 induces cytotoxicity to both CLL-derived cell lines Mec-1 and OSU-CLL, as well as in primary CLL cells, while illustrating alleviated toxicity in normal B and T cells (Figure 1B, C). In contrast to the highly successful BTK and PI3 kinase inhibitors which inhibit BCR signaling proximally, OSU-T315 specifically inhibits down-stream AKT and ERK signaling cascades without affecting proximal BCR or focal-adhesion signaling. In particular, external stimuli through BCR or CD49d, by which triggers survival signal and protects CLL from ankoisis via AKT/ERK activation, is potently inhibited by OSU-T315. Downstream anti-apoptotic molecule Mcl-1 is suppressed upon OSU-T315 treatment, which is later accompanied by increased Caspase 3/7 activity. Further study indicates that OSU-T315 mediated cell death is Caspase dependent. In vivo efficacy of OSU-T315 is supported by a therapeutic study in the TCL1 mouse model of CLL, from which demonstrates improvement in total counts of white blood cells and reduced splenomegaly, thereby contributing to enhanced overall survival following OSU-T315 oral treatment regimen (Figure 2). Together, our findings suggest the innovative dual-targeted approaches by suppressing both AKT and ERK pathway in CLL therapy through inhibiting ILK, and provide an alternative agent for potential therapeutic development in CLL.

Disclosures:

No relevant conflicts of interest to declare.