bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gene-Expression Profile of Steroid-Refractory Acute Graft-Versus-Host Disease.
Authors: Verner Jan , Kabathova Jitka , Zdrahal Zbynek , Tomancova Alexandra , Volejnikova Jana , Brychtova Yvona , Pospisilova Sarka , Mayer Jiri 
Published: October 2019

Abstract
Abstract 4549
Steroid-refractory acute graft-versus-host disease (aGVHD) remains to be the major complication of allogeneic hematopoietic stem cells transplantation (allo-HSCT). aGVHD is characterized by tissue injury and is associated with high morbidity and mortality, most notably in cases of severe course and/or resistance to corticoids. aGVHD appears usually within the first 100 days after transplantation. This study should lead to identification of molecular patterns characterizing patients’ sensitivity or resistance to corticoid therapy. We performed an oligonucleotide microarray analysis (whole genome Human OneArray, Phalanx) of 13 patients with hematological malignancies who received allo-HSCT. Peripheral blood samples were collected at the time of aGVHD manifestation and subsequently mononuclear cells were isolated (Ficoll-Paque gradient). In total, five patients with resistance, and eight patients with sensitivity to corticoids were included in the analysis. The significance analysis of micorarrays (SAM) revealed a set of differentially expressed genes: GBP5 (guanylate binding protein 5), PRG3 (proteoglycan 3), CDK5RAP2 (CDK5 regulatory subunit associated protein 2), and, especially, ARG1 gene (arginase, liver) whose expression was significantly higher in the group of resistant patients (with 4.89-fold change). As published earlier, arginase is released from human granulocytes during purulent inflammatory reactions and induces a suppression of T-cell proliferation and cytokine synthesis (Munder et al., 2006). These findings might confirm our hypothesis that expression levels of ARG1 gene could reflect the severity of aGVHD. ARG1 gene was detected apparently in a fraction of immature forms of granulocytes which are released into the peripheral blood after allo-HSCT. Thus, both immature granulocytes and mononuclear cells are then isolated using Ficoll-Paque gradient. To validate arginase as a marker of unfavorable grade of aGVHD, it is necessary to examine expression profile of ARG1 gene in neutrophils. Therefore, it is important to separate both mature and immature forms of neutrophils (using different method than Ficoll gradient), as well as examine gene expression profile of a down-regulated CD3ζ chain in T-cells. Down-regulation of CD3ζ chain is due to arginase-mediated depletion of arginine in the T-cell environment (Munder et al., 2006). We assume that ARG1 gene expression profile might have a potential to influence patients′ response to corticoid therapy of unfavorable grade of aGVHD. This study is in progress and further analyses will be focused on monitoring of ARG1 and CD3ζ chain expression levels in a larger cohort.
Supported by IGA MZ CR NS9683-4/2008 and MSMT CR MSM0021622430.

Disclosures:
No relevant conflicts of interest to declare.
Comparative Analysis of Single-Institute Transplantation From Unrelated Cord Blood and Related Adult Donors In Adults with Hematologic Malignancies.
Authors: Sun Zimin , Liu Huilan , Geng Liangquan , Wang Xingbing , Ding Kaiyang , Zhou Chengyang , Tang Baolin , Tong Juan , Yao Wen , Wang Zuyi 
Published: October 2019

Abstract
Abstract 4570
We compared the clinical outcomes of allogeneic hematopoietic stem-cell transplantation among 126 adults with hematologic malignancies: 40 received unrelated cord blood transplantation (CBT) and 86 received related bone marrow transplantation (BMT) and/or peripheral blood stem cells transplantation (PBSCT). All patients received myeloablative transplantations which performed from 2001 through 2010.Recipients of unrelated CBT were younger (median, 23.5 vs. 30 years of age; P<0.001) and had more advanced disease at the time of transplantation (68% vs. 34%, P<0.001) than recipients of related BMT/PBSCT. All related transplants were HLA matched, whereas 77 percent of CBT grafts were HLA mismatched (P<0.001). The median number of nucleated cells infused was 0.37×108 per kilogram of the recipient's body weight for CBT and 5.39×108 per kilogram for BMT/PBSCT recipients (P<0.001). Neutrophil recovery was significantly delayed after CBT (median, 19 vs. 12 days post transplantation, P<0.001), however, overall engraftment rates were almost the same for both grafts. Multivariate analysis demonstrated no apparent differences in transplantation-related mortality (TRM) (33% in CBT and 21% in BMT/PBSCT recipients), relapse rate (15% in CBT and 15.1% in BMT/PBSCT recipients), the incidence of grades III to ‡W acute graft-versus-host disease (aGVHD) (7.5% in CBT and 5.8% in BMT/PBSCT recipients), 2 years disease free survival (DFS) (52.5% in CBT vs. 62.8% in BMT/PBSCT recipients) and over survival (OS) (62.5% in CBT vs. 67.4% in BMT/PBSCT recipients) between both groups, but extensive chronic GVHD (cGVHD) was lower in CBT recipients (2.5% vs. 17.4%). These data suggest that unrelated CBT could be as safe and effective as matched related BMT or mobilized PBSCT for adult patients with hematologic malignancies.

Disclosures:
Sun: International Cooperation Research Fund of Anhui Provincial Scientific and Technologic Committee (08080703026): Research Funding; Fund of the Key Medical Project of Anhui Provincial healthy department (2010A005): Research Funding; Anhui Provincial “115” Industrial Innovation Program: Research Funding.
Encouraging Results of Reduced-Intensity Conditioning (RIC) Followed by Hematopoietic Stem Cell Transplantation (HSCT) from an Alternative Donor for Acute Lymphoblastic Leukemia (ALL) In Children.
Authors: Inoue Masami , Yasui Masahiro , Tokimasa Sadao , Sawada Akihisa , Sato Maho , Kondo Osamu , Yamada Kayo , Kawa Keisei 
Published: October 2019

Abstract
Abstract 4572

Background:
RIC has been applied mainly for elderly patients and patients with comorbidity to expand opportunity for such adult patients to undergo HSCT. The burden of late effects after HSCT followed by myeloablative conditioning is not negligible in pediatric patients. RIC should be introduced to avoid not only treatment related morbidity (TRM) but also late effects for children even if they are not ineligible for myeloablative transplantation. However, enhanced graft-versus-leukemia (GVL) effect by choosing an alternative donor in the setting of RIC-HSCT for pediatric ALL hasn't been clarified, so far.

Methods:
We retrospectively analyzed 22 children with ALL who underwent RIC followed by HSCT from an alternative donor between 2001 and 2008 in our institute.

Results:
The median age of 22 patients at HSCT was 9 years (range one year - 19 years). 8 patients in CR1, two in CR2 and 12 beyond CR2 underwent HSCT from an alternative donor followed by Flu 180mg/m2 and LPAM 140 mg/m2 containing RIC. 8 patients who relapsed after myeloablative HSCT and 5 patients with Ph1-ALL were included. Unrelated bone marrow (BM) in 7 patients and unrelated cord blood in 4 were transplanted. 11 patients were transplanted from a haplo-identical parent (8, BM; 3, selected CD34 positive peripheral blood mononuclear cells). Engraftment was confirmed in 21 transplants out of 22 (95.4%). Death of TRM within 100 days after transplant was observed in 3/22 (13.6%). Acute graft-versus-host disease (GVHD) grade IV wasn't observed. Acute GVHD grade II and III occurred in 8/21 (38.1%) and 6/21 (28.6%), respectively. Chronic GVHD was seen in 8/16 (50%). 14 patients (63.6%) out of 22 are surviving after RIC-HSCT for median 36 months (range, 24 – 100 months). 9 patients (90%) out of 10 in CR1/CR2 and 5 (41.7%) out of 12 beyond CR2 are surviving.

Conclusion:
These results suggest RIC-HSCT from an alternative donor for pediatric ALL can be performed safely and similar outcome can be expected compared to myeloablative transplantation.

Disclosures:
No relevant conflicts of interest to declare.
Allogeneic Stem Cell Transplantation for Patients with AML and FLT-3 Mutations.
Authors: Schleuning Michael , Judith Dirk , Heshmat Mansoor , Burlakova Irina , Jedlickova Zuzanna , Salm Elisabeth , Baurmann Herrad , Schwerdtfeger Rainer 
Published: October 2019

Abstract
Abstract 4573
Patients with acute myeloid leukemia (AML) with FLT-3 mutations have an extremely high risk of relapse after conventional chemotherapy. The role of allogeneic stem cell transplantation (SCT) for this patient cohort has been discussed controversially in recent years. This retrospective analysis reports our cumulative experience in a cohort of 42 consecutive patients (age 17–70, median 51 years) allografted for FLT-3 positive AML in a single centre. In more than 80% a FLAMSA-RIC based conditioning regimen was used, in 5% BCNU/Melphalan/Fludarabine, and in 14% conventional radiation- or Busulfan- based regimens. Most patients received mobilized peripheral blood stem cells as graft and 10 patients had a sibling and 32 an unrelated donor (MUD), respectively. Half of the patients were allografted in complete remission and twenty with active, mostly refractory disease. With a median follow-up for surviving patients of nearly 2 years (range 64 – 1746 days) the Kaplan-Meyer procedure estimates a 48% probability of survival at 2 y after transplantation. Interestingly, there is no difference what so ever in survival if patients had an identical sibling donor or a MUD. Similarly, neither patient age below or above the median, nor the applied conditioning regimen did affect the outcome. The only significant variable for improved survival was being in complete remission at transplantation with a 2-year overall survival probability of 60% as compared to 30% for patients with active disease. Thirteen patients (31%) relapsed after allografting, which is substantially lower as to what is reported after conventional chemotherapy. Three of these patients could be salvaged by a second transplant, whereas 10 patients finally died from leukaemia. Non relapse mortality was 24% with 2 patients dying from acute GVHD, 7 from infections and 1 from suicide, despite being well physically. In conclusion, our data support the notion that allogeneic SCT is a highly effective treatment option for patients with AML and FLT-3 mutations and that, if the patient is eligible, it should be undertaken whenever possible in 1. complete remission. However, even patients with primary induction failure have a reasonable chance to be salvaged by allogeneic SCT.

Disclosures:
No relevant conflicts of interest to declare.
Successful Rescue of Childhood Acute Leukemia In Non-Remission by Non-T-Cell Depleted HLA-Haploidentical Hematopoietic Cell Transplantation.
Authors: Kobayashi Shogo , Kikuta Atsushi , Ito Masaki , Sano Hideki , Mochizuki Kazuhiro , Akaihata Mitsuko , Ohto Hitoshi , Hosoya Mitsuaki 
Published: October 2019

Abstract
Abstract 4582
Refractory acute leukemia who do not achieve second or subsequent complete remission (CR) have an extremely poor prognosis. Non-T-cell depleted (TCD) HLA-haploidentical hematopoietic cell transplantation (haplo-HCT) is a form of adoptive cellular therapy that has a high degree of efficacy in hematologic malignancies. The major problems of non-TCD haplo-HCT are severe graft-versus-host disease (GVHD), graft failure (GF) and high-risk of early death. We conducted non-TCD haplo-HCT for children with acute leukemia in non-remission as a novel therapeutic strategy.
Five consecutive children under 15 (0-13) years old with hematological malignancies underwent non-TCD haplo-HCT from family donors between November 2002 and July 2010 at Fukushima Medical University Hospital. All patients were in non-remission at transplantation. One donor was mother and the other 4 donors were fathers of the patients. One patient was in 3 loci, and 4 patients were mismatched 4 loci of the allele level for HLA-A, -B, -C and -DRB1. The blast count in the marrow before transplantation was 80%, 75%, 34%, 10% and 96%, respectively. There were two acute myelogenous leukemia and three acute lymphoblastic leukemia. There were two refractory diseases to chemotherapy and three relapses after HSCT. All of them received myeloablative conditioning (2 Busulfan based, 3 TBI based). Type of graft were bone marrow in one and peripheral blood stem cell in 4. Total of 9.9 ×108/kg of nucleated bone marrow cells were infused in the patient received BMT. The median number of TNC and CD34+ cell doses were 11 (8-21.4)×108/kg and 7.9 (6.5-8.8)×106/kg in PBSCT, respectively. GVHD prophylaxis were combination of tacrolimus, methotrexate was given in one patient, combination of tacrolimus, methotrexate and prednisolone in one, combination of tacrolimus, methotrexate, prednisolone, and antihuman thymocyte immunoglobulin (ATG) in 3.
All patients achieved primary engraftment at median of 13 days (11 -15) and achieved complete remission. Five patients developed acute GVHD, with grade 1 in two patients, grade 2 in two patients, grade 3 in one patient. Chronic GVHD occurred in 2 of 4 evaluable patients. Infectious complications including 2 CMV reactivation, 1 CMV-IP, 1 invasive aspergillosis, 1 candida sepsis were observed. One patient died from CMV-IP in remission, remaining 4 patients survived disease-free + 4, + 9, + 10 and + 94 months after non-TCD haplo-HCT, respectively.
These preliminary results suggested that non-TCD haplo-HCT is effective strategy in children with refractory leukemia.

Disclosures:
No relevant conflicts of interest to declare.
Oral Supplementation with Omega3 Fatty Acids Inhibits NFkB Activation In Chronic Lymphocytic Leukemia (CLL) Cells.
Authors: Ballester Oscar F. F , Fahrmann Johannes , Witte Theodore , Ballester Gabriela , Hardman W. Elaine 
Published: October 2019

Abstract
Abstract 4607

Introduction:
Nuclear factor kappa B (NFkB) is a critical transcription factor involved in the growth and survival of CLL cells. NFkB is recognized as an important target for the development of novel therapies for the treatment of various malignancies. In vitro and in experimental animal models, OMEGA-3 fatty acid (O3FA) supplementation has been shown to inhibit NFkB activity.

Patients and Methods:
Patients with early stage CLL (Rai stages 0-II) who required no therapy, where accrued to this phase I-II trial. O3FA supplements were given for a total of 12 months at doses ranging from 2250 mg (EPA plus DHA), escalated to 4500 mg and 6750 mg per day as tolerated. NFkB activity was measured in peripheral blood samples after separation of mononuclear cell by gradient centrifugation and expressed as luminescence units/μ g of protein. Baseline and multiple serial samples were obtained during the study period. In-vitro cytotoxicity assays to doxorubicin were conducted using standard LD50 methods. Compliance was monitored by analysis of red cell and lymphocyte membrane lipid composition by gas chromatography.

Results:
Fifteen patients have been accrued to the trial, 8 of them have currently completed the planned 12 months of the study period. No significant clinical changes in disease activity were noted. O3FA was well tolerated. Supplementation resulted in a dose-dependent increase of O3FA composition of red cell and lymphocyte membranes in a dose dependent manner. At baseline, CLL patients had NFkB above the range observed in normal controls (2.05 × 104 to 2.32 × 105 NFkB lum units/μ g). The median value in CLL patients at baseline was 11.60 × 106 NFkB lum units/μ g (range 0.9 × 105 to 23.12 × 106). Among 5 patients with the highest baseline levels of NFkB, a decrease in NFkB activity ranging from 0.02 to 0.19 of the baseline value, was noted at the 2 higher doses of O3FA supplementation. Similar results were seen in patients with relatively lower levels of baseline NFkB activity (0.9 × 105 to 2.96 × 106 lum units/μ g). In vitro, significant doxorubicin cytotoxicity (>50%) was noted in samples obtained during supplementation, at μ gM concentrations which produced no detectable cell kill in baseline samples.

Conclusions:
O3FA supplementation resulted in significant inhibition of NFkB activity in leukemic cells from patients with CLL. In-vitro, after O3FA supplementation CLL cells became more sensitive to doxorubicin. Preliminary analysis of whole genome micro arrays revealed significant down-regulation of multiple genes associated with O3FA supplementation.

Disclosures:
No relevant conflicts of interest to declare.
Histone Deacetylase Activity In Chronic Lymphocytic Leukemia.
Authors: Kafeel Muhammad I. , Avezbakiyev Boris , Chen Chi , Sun Yiwu , Rathnasabapathy Chenthil , Kalavar M. , He Zili , Burton Jack , Lichter Stephen M. , Wang Jen-Chin 
Published: October 2019

Abstract
Abstract 4622
Elevated histone deacetylase (HDAC) enzyme levels have been described in patients with carcinoma and leukemia. Using HDAC inhibitor (HDACi) to treat carcinoma has been promising and is currently undergoing intense research. In order to be better in using HDACi to treat chronic lymphocytic leukemia (CLL), HDAC isoenzyme levels were measured in 32 patients with CLL and compared with 17 normal volunteer controls. Peripheral blood CD20+ cells from patients with CLL and normal volunteer controls were isolated by using Progenitor Cell Isolation Kits (Miltenyi Biotec, Auburn, CA). Isolated CD20+ cells were confirmed by flow cytometry studies to have more than 92% purity. Patient and control CD20+ cells were then lysed in denaturation buffer. Total RNA was extracted, then cDNAs were prepared and quantitative RT-PCR was performed using premixed primer and FAM- and TAMRA-labeled probes obtained from ABI. The results showed: 1) HDAC1, -3, -6, -7, -9, and -10 and SIRT1, -2, and -6 were significantly over-expressed, suggesting that, in CLL, elevated HDAC activity are not restricted to one class. Therefore, HDAC inhibitor therapy may need to be directed to more than one specific class of HDAC; 2) Patients with ZAP70+ were associated with more advanced Rai clinical stage and higher HDAC activity than Zap70– patients and HDAC activity were correlated to the expression levels of CD44 and Pin1, suggesting that higher HDAC values may indicate a poor prognosis and more advanced disease stage.

Disclosures:
No relevant conflicts of interest to declare.
Fludarabine Versus No-Treatment In the Patients with CLL Stage A
Authors: Cahn Jean-Yves , Ivanaj Arben , Pivot Xavier , Dobi Erion , Pulluqi Polikron , Xhumari Pal 
Published: October 2019

Abstract
Abstract 4639

Background.
Fludarabine is an effective treatment for chronic lymphocytic leukemia (CLL) alones and in combined terapy in untreatment CLL. The standard recomandation for the stage A Binet, is to delay any treatement until progression. We compared the benefit of early exposure to fludarabine versus no-treatment in the patients with CLL stage A.

Methods.
Between 2000 and 2004, we randomzed 77 patients with CLL stage A to be treated; fludarabine 25 mg/m2, administered intravenously daily for 5 days every 28 days versus no treatment until disease progression. The treated group with received the assigned treatment for a maximum of 6 cycles. Both groups evaluated evry 2 month diseases progression. The NCI-WG response criteria for CLL was refered for analized result.

Results.
Among the 38 patients treated by fludarabine. Complet response and parcial response were observed 21 % and 74 % of cases respectively. The median Progression Free Survival (PFS) was 32 month in the treated group versus 31.5 months in the 39 patients of control group, (p 0.9). The median survival were 124 and 122 months respectively, p 0.7. Existing deletion p53, high risk stage A CLL had no impact on the result. The commonest grade III-IV toxicities were neutropenia (12 %) and trombocytopenia (11 %) in the tretaed group.

Conclusions.
Fludarabine did not prolong PFS and survival in patients with stage A CLL. Despite the low tolerability of fludarabine there is no need to provide an early exposure in patients with stage A CLL.

Disclosures:
No relevant conflicts of interest to declare.
Activation of Paired-Homeobox Gene PITX1 by Del(5)(q31) In T-Cell Acute Lymphoblastic Leukemia (T-ALL)
Authors: Nagel Stefan , Venturini Letizia , Przybylski Grzegorz K. , Grabarczyk Piotr , Schneider Björn , Meyer Corinna , Kaufmann Maren , Schmidt Christian A. , Scherr Michaela , Drexler Hans G. , MacLeod Roderick AF 
Published: October 2019

Abstract
Abstract 4644
Aberrant activation of various oncogenes, including homeobox genes encoding fundamental transcription factors, during T-cell development contributes to T-cell acute lymphoblastic leukemia (T-ALL). Neoplastic chromosome rearrangements are known to deregulate Antennapedia-class homeobox genes of the NKL-family (TLX1, TLX3, NKX2-5) and HOXA-cluster genes of the Extended-Hox-family. Following analysis of T-ALL cell lines and primary cells, we describe leukemogenic involvement of a third homeobox gene group, the Paired (PRD)-class. Ascertainment was performed in an early stage-arrested T-ALL cell line (LOUCY) which revealed chromosomal deletion at 5q31, removing the downstream regulatory region of the PRD-homeobox gene PITX1. Comparative expression analysis confirmed ectopic PITX1 expression, consistent with aberrant activation by del(5)(q31) which removes a STAT1 binding site. STAT1 mediates repressive IL2-STAT1 signaling, implicating IL2-pathway avoidance as a possible activation mechanism. Furthermore, we detected expression of the physiologically similar PITX2 in 11/24 (46%) T-ALL cell lines, 5 with genomic PITX2 gains. Among primary T-ALL samples, 2/22 (9%) – both pediatric pre-T-ALL - ectopically expressed PITX1 but not PITX2. Forced expression of PITX1 by lentiviral transduction of JURKAT cells and subsequent analysis by expression profiling, prompted upregulation of RUNX2 and JUN and inhibition of RUNX1 and NKX3-1, indicating impaired T-cell differentiation. Taken together, our data show leukemic activation of PITX1, a novice PRD-class homeobox gene in the T-ALL repertoire, which may promote leukemogenesis by inhibiting differentiation.

Disclosures:
No relevant conflicts of interest to declare.