bluuebunny/crossref_metadata_2025_split huggingface dataset:
Phase I/II Study of Escalating Doses of Idarubicin in Combination with Methotrexate, Vindesine and Prednisolone in Patients with Primary Central Nervous System Lymphoma: Results of the GOELAMS LCP99 Multicenter Trial.
Authors: Olivier Gaelle , Gardembas Martine , Delwail Vincent , Le Mevel Annie , Escoffre-Barbe Martine , Brion Annie , Boulet Jean Michel , Solal-Celigny Philippe , Ingrand Pierre , Colombat Philippe 
Published: October 2019

Abstract
Abstract 2694
Poster Board II-670

Introduction:
This study was performed to determine the maximum-tolerated dose (MTD) and the recommended dose (RD) of idarubicin in combination with methotrexate (MTX), vindesine (VDS) and prednisolone (PRED) given to previously untreated patients (pts) (60–70 years-old) with primary central nervous system lymphoma (PCNSL).

Patients and Methods:
MTX and VDS were respectively given at the fixed dose of 3 g/m2 (6-hour intravenous [IV]) and 3 mg/m2 IV both on day 1. PRED was given at the fixed dose of 60 mg/m2 (IV or per os) on day 1 to 5. Idarubicin was administrated IV on day 1 with doses starting at 12 mg/m2 and escalated by 2 mg/m2 increments until MTD. Treatment was repeated every 3 weeks, three times. G-CSF was used. Intrathecal chemotherapy (MTX 15 mg, cytarabine 40 mg ) was performed on day 1. Subsequent whole brain irradiation (30 Gy + 10 Gy on residual mass) was performed. Limiting toxicity was defined as grade 4 neutropenia for more than 7 days or thrombopenia grade 4 or extra haematological toxicity more than grade 2.

Results:
Between 2000 and 2005, 35 immunocompetent pts (19 men, 16 women) with a new diagnosis of PCNSL received 87 cycles at four different idarubicin dose levels. At dose level 1 (12 mg/m2) and level 2 (14 mg/m2), only one of six developed a limiting toxicity. About 3 cases, no limiting toxicity was observed at level 3 (16 mg/m2). Three of six developed a limiting toxicity at level 4 (18 mg/m2) which was considered as MTD. Fourteen patients were enrolled at level 3 to confirm the data. At level 3, main haematological toxicities were neutropenia grade 3 (9%) or grade 4 (40%) and main extra haematological toxicities were infectious disease (5 % grade 3 or 4) and renal failure (grade 2: 9%). Median survival time was 385 days (IC95% 136–679).Overall response rate after radiotherapy was 65.7 % (15 CR, 7 PR).

Conclusion:
Our study suggests that the RD of idarubicin administrated with MTX, VDS and PRED, should be 16 mg/m2 on day 1 every 21-day cycle.

Disclosures:
Delwail: Innate Pharma: Honoraria.
CD23 Receptor Positivity Has No Prognostic Implication in Chronic Lymphocytic Leukemia.
Authors: Aghalar Jahan , Chu Charles , Damle Rajendra N , Tsao Che-Kai , Kohn Nina , Kaufman Matthew 
Published: October 2019

Abstract
Abstract 4397

BACKGROUND
Chronic Lymphocytic Leukemia (CLL) phenotypically expresses CD23, although the percentage of positive cells measured by flow cytometry is variable. We sought to analyze whether the percent of CD23 positive cells in the CLL clone correlates with time to treat (TTT), overall survival (OS) and prognostic markers CD38, ZAP-70, and IGHV mutation status.

METHODS
We retrospectively analyzed the flow cytometry data of 332 CLL patients on the gated population of cells that were CD5 and CD19 positive. Percentage positivity for CD23, CD38, and ZAP-70 was noted. CD38 and ZAP-70 were considered positive at cut-offs of >= 30% and >=20%, respectively. CD23 was considered negative at <30% and positive at >= 30%. IGHV sequence was determined from cDNA and then compared to germline to assess mutation status using IMGT/V-QUEST.
The distributions of time from diagnosis until start of treatment and overall survival were stratified by CD23 positivity, estimated using the product limit method, and compared using the log rank test. Those who had expired without treatment or were alive and not treated at this time point were censored in the TTT analysis. Those who were still alive were censored in the OS analysis. Associations of CD23 positivity with IGHV mutation status, ZAP-70, and CD38 positivity were examined using the chi-square test.

RESULTS
Out of 332 patients, 25 had diminished CD23 expression (<30%) whereas 307 had normal CD23 expression (>30%). There was no difference in time until start of treatment or overall survival based on CD23 %positivity. CD23 %positivity showed no associations with IGHV mutation status, ZAP-70 or CD38 positivity.

CONCLUSION
CD23 percent positivity has no prognostic significance in CLL. There is no correlation between CD23 percent positivity and poor prognostic markers such as CD38, ZAP-70, or IGHV mutation status.

Disclosures:
No relevant conflicts of interest to declare.
Deacetylase Inhibitor Cocktails Provide Striking Synergistic Interactions in Human Leukemia Cells.
Authors: Cea Michele , Cagnetta Antonia , Fruscione Floriana , Bruzzone Santina , Zoppoli Gabriele , Garuti Anna , Moran Eva , Grillo Valentina , Ferrando Fabio , Casciaro Salvatore , Pierri Ivana , Gobbi Marco , Ballestrero Alberto , Patrone Franco , Nencioni Alessio 
Published: October 2019

Abstract
Abstract 4404
Cancer cells almost invariably exhibit aberrant histone deacetylase (HDAC) activity leading to changes in chromatine structure, altered gene expression, poor differentiation, impaired apoptosis and increased proliferation. Accordingly, virtually all the HDAC inhibitors currently available show some degree of antitumor activity in preclinical cancer models and several of these compounds are currently under investigation or already approved for the treatment of human malignancies. Such is the case of the hydroxamic acid derivative suberoylanilide hydroxamic acid (Vorinostat, Zolinza), approved for the treatment of cutaneous T cell lymphomas. Sirtuins are a large family of deacetylases characterized by a unique, NAD+-dependent enzymatic mechanism. In addition to their established role in metabolism and longevity, recent evidence points to an emerging role for sirtuins in carcinogenesis. In the attempt to identify drug combinations that would increase the activity of traditional HDAC inhibitors we have explored the combination of valproic acid (VA) and butyrate (BU) with the sirtuin inhibitors cambinol and sirtinol in primary B-cell chronic lymphocytic leukemia (B-CLL) cells (n=35), acute myelogenous leukemia (AML) cells (n=10) and leukemia cell lines. Cell viability was assessed by propidium iodide staining and flow cytometry. Combination indices were determined using the median-effect method. In leukemia cells, exposure to sirtuin inhibitors synergistically increased VA and BU mediated cytotoxicity. Conversely, these drugs were poorly active and failed to show any cooperation in healthy cells, including peripheral blood mononuclear cells and fibroblasts, suggesting a cancer-specific mode of action. Similar results were obtained by combining VA or BU with the Nampt inhibitor APO866, which reduces intracellular NAD+ levels and thereby prevents sirtuin activity. Remarkably, SIRT1 and SIRT6 inhibition per se did not seem to account for cell demise upon HDAC inhibition since expression of a dominant negative SIRT1 isoform or RNA interference-mediated SIRT6 silencing failed to increase VA and BU activity. Our data indicate a specific requirement by leukemia cells for sustained sirtuin activity when classical HDACs are inhibited. This feature is suitable to be therapeutically exploited by combining sirtuin inhibitors or APO866 with classical HDAC inhibitors especially for the treatment of hematological malignancies.

Disclosures:
No relevant conflicts of interest to declare.
Gene Profiling Mechanisms of Cell Density-Dependent Growth in Myeloid and Lymphoid Leukemia Cell Lines.
Authors: Murohashi Ikuo , Ihara Noriko 
Published: October 2019

Abstract
Abstract 4420
Normal hematopoietic stem cells have been shown to be maintained through interaction with their environmental niches, such as osteoblastic and endothelial ones. The growth of leukemic cells has been shown to be stimulated by environmental niches (paracrine growth) or by cell-to-cell interaction or excreted factors of leukemic cells (autocrine growth). The growth of myeloid (MO7-E and HL-60) and lymphoid (Raji, U-266, Daudi and RPMI-1788) leukemia cell lines cultured at various cell densities in serum free medium (Sigma H 4281) with 1% BSA was evaluated. The cells cultured at higher cell densities (cultured cell densities ≥a 105/ml) showed logarithmic linear increases in cell number, whereas those at lower cell densities (cultured cell densities □… 104/ml) ceased increasing cell number. Supernatants of myeloid leukemia cells stimulated the growth of autologous clonogenic cells, but not those of lymphoid leukemia cells. Neutralizing antibodies (Abs) against various hematopoietic growth factors failed to inhibit cell growth except for anti-VEGF, which significantly decreased HL-60 leukemia cell growth. To clarify the nature of the cultured cell density on the growth of leukemia cells, leukemia cells were cultured at higher cell density (group H, cultured cell densities of 106/ml) or at lower cell density (group L, cultured cell densities 104/ml). After culture of 3-, 6-, 10-, and 24-hr, cells were serially harvested and total cellular RNA was extracted. Gene transcript levels were determined by using Real-Time PCR. Gene transcripts examined in the present study were as follows: polycomb (Bmi1), Hox (HOXA7, HOXA9, HOXB2, HOXB4, Meis 1), Caudal-related (CDX2, 4), Mef2c, c-Myb, Wnt (Wnt 3a, Wnt 5a, β-Catenin, β-Catenin, N-Cadherin), Notch (Notch-1, -2, -3 and Jagged-1, -2), CKI (p14, p15, p16, p18, p21, p27, p57), growth factor (VEGF, IGF-1, -2, Ang-1, -2, SDF-1), growth factor receptor (Flt-1, KDR, neurophilin-1, IGF-1R, Tie-1, -2, CXCR4), and growth related (c-Myc, CyclinD1, Foxo3a) genes. p18 and p21 gene expression was higher in group L compared with group H in two and all five groups, respectively. In contrast, p14 gene expression was higher in group H compared with group L. Any of the p15, p16, p27 and p57 genes was deleted. VEGF gene expression levels at 1-3- hr culture were higher in group H compared with group L. HOX, Meis 1 and Mef2c gene expression levels at 1- to 10- hr culture were higher in group H compared with group L. At 24-hr cultures, transcripts of myeloid and lymphoid cell lines for Bmi-1, Wnt-3a, and β-Catenin were higher, and those of lymphoid cell lines for Notch 1, 2, and 3 were higher in group H compared with group L. Taken together, our present results favor the conclusions that genes related to growth factors and transcription factors are sequentially and differentially expressed through cell-to-cell interaction and excreted autocrine growth factors of leukemia cells.

Disclosures:
No relevant conflicts of interest to declare.
Hyper-Methylation DAZAP2 May Suppress Its Expression in Specific Subtypes of Myeloma.
Authors: Hu Wei-Xin , Qu Qiang , Li Jiang , Ren Wei 
Published: October 2019

Abstract
Abstract 4421
Most malignant features of cancer cells are triggered by activated oncogenes and the loss of tumor suppressors due to mutation or epigenetic inactivation. We previously reported that DAZAP2 was down-regulated in newly diagnosed myeloma (MM) and this may influence MM cell growth and survival. This study was undertaken to evaluate the mechanism of down-regulation DAZAP2 and determine the methylation status and loci of its promoter by using bisulphite genomic-sequencing (BGS) method in KM3 myeloma cell line. Two islands with rich GC box in the promoter of DAZAP2 gene were identified and amplified by using bisulfite-sequencing PCR (BSP). Island 1 spans -472 to -247 including 9 CpG sites (CpGs) and island 2 covers -226 to -13 including 23 CpG sites. The ratio of methylated CpGs in two CpG islands was 46.25%. The above sequences were demethylated and inserted into a pGL2-basic vector. COS-7 cells were transfected with recombinant plasmids and the activity of luciferase was evaluated. The results showed that the CpG island 1 had a weakly transcriptional activity, whereas the CpG island 2 had a strong transcriptional activity (2.38 folds compared with the positive control). The other sequence which covered CpG island 1 and 2 showed a remarkable activity (15.1 folds compared with the positive control). These data indicated that CpG island 2 of DAZAP2 gene may be hypermethylated and suppressed its expression in MM cells. We further correlated DAZAP2 expression with normal plasma cells and malignant myeloma cells, as well as the molecular subtypes which the dataset includes 8 genetic subtypes (MY, PR, LB, MS, HP, CD-1, CD-2, and MF) from 351 newly diagnosed myeloma cases (Zhan, 2006). Interestingly, we extended and confirmed our previous discovery that DAZAP2 was significantly down-regulated in MM cells by using a large uniform dataset (P = 0.004). The low expression of DAZAP2 was especially significant in the subgroups of MY, PR, LB, HP, and CD-1. This study warrants further investigation of DAZAP2 and its potential role in myeloma.

Disclosures:
No relevant conflicts of interest to declare.
Change of the Notch1 Expression Level in Lymphoid Malignancies.
Authors: Zheng Haitao , Yang Lijian , Chen Shao-Hua , Zhong Liye , Li Yangqiu 
Published: October 2019

Abstract
Abstract 4425
Notch gene family encodes single pass transmembrane receptors able to transducer intercellular signals involved in cell-fate determination. Notch signaling is the leading causes to the development of malignant tumors. Notch pathway plays a role in haematopoiesis affecting both stem cells and committed progenitors. The expression of Notch1 in thymic stromal cells and in developing thymocytes is consistent with Notch function. Notch1 is turn on before T-cell commitment along the lymphoid development; a variety of model systems indicates that Notch1 signaling is required for the earliest stages of T-cell commitment in the thymus and that, in its absence, cells develop toward the B-lineage. Overexpression of Notch1 gene was found in T-ALL, however, little is known the expression pattern of Notch1 gene in the different lymphoid malignancies. In the present study, the expression level of Notch1 gene was analyzed in patients with lymphoid malignancies. Real-time PCR with SYBR Green I technique was used for detecting Notch1 gene expression level in peripheral blood mononuclear cells of 43 patients with lymphoid malignancies [including 7 cases with T-ALL, 6 cases with T-NHL, 9 cases with B-ALL, 5 cases with B-CLL, 16 cases with B-NHL] and 20 healthy individuals. β2-microglobulin gene (β2M) was used as an endogenous reference. Relative changes in Notch1 gene expression level were used by the 2-ΔCt×100% ×100% method. The expression level of Notch1 in both T-cell lymphoid malignancies group (Median: 0.897%) and B-cell lymphoid malignancies group (Median: 0.726%) was significant higher than those in the healthy control (Median: 0.288%) (P<0.01, P<0.01). Moreover, the expression level of Notch1 was different in the five types of lymphoid malignancies. That is, the highest in the B-CLL group, secondary in the T-NHL group, the lowest in the B-NHL group. And, it is not significant different between the B-NHL and healthy control groups. In conclusions, Overexpression of Notch1 gene was found not only in T-ALL, but also in T-NHL and B-cell leukemia and suggests that the abnormal overexpression pattern of Notch1 might play an important role in the pathogenesis of lymphoid malignancies. Down-regulation of Notch1 might be considered as target therapeutic strategy for lymphocytic malignancies.

Disclosures:
Zheng: The study was supported by grants from National Natural Science Foundation of China (No. 30871091): Research Funding. Yang:The study was supported by grants from National Natural Science Foundation of China (No. 30871091): Research Funding. Chen:The study was supported by grants from National Natural Science Foundation of China (No. 30871091): Research Funding. Zhong:The study was supported by grants from National Natural Science Foundation of China (No. 30871091): Research Funding. Li:The study was supported by grants from National Natural Science Foundation of China (No. 30871091): Research Funding.
Hemophilia in the Peruvian Social Security System: Report of Five Centers.
Authors: Chumpitaz Gloria , Cauvi Fernando , Navarro Juan Ramon , Pedraza Karina 
Published: October 2019

Abstract
Abstract 4446
The Hemophilia Unit of the Hematology Department of the National Hospital Edgardo Rebagliatti Martins- ESSALUD, is one of the most important Hemophilia Centers through the country, that assists patients not only in its jurisdiction but also a great amount referred from other institutions due the complexity of their treatment, like acquired inhibitor disorders as well as orthopedic and mayor cardiovascular surgeries and severe hemorrhages.
The Hemophilia ESSALUD system is compounded of centers in Lima (02), Callao (01) and provinces (05) (Arequipa, Chiclayo, Trujillo, Piura and Cuzco).
The range of patients with new diagnosis is about 5 to 20 patients per year.
The prevalence of Hemophilia in the last 5 years, accounts 83.38% patients of the global amount with coagulation disorders. Of the 331 patients in treatment, 228 (68.8%) have Hemophilia A. Considering the classification of status severity, 12.5% patients of this group belong to mild status hemophilia, 39.47% patients to moderate status, and 41.43% to severe status.
In relation to Hemophilia B, we account 48 (14.5%) patients; 5.2% corresponds to mild status, 23.68% moderate status and 52.63% severe status.
The rest of the patients have other disorders of coagulation such as Von Willebrand disease and rare coagulation factor deficiency (V, VII, XI and XII).
According to the age group distribution for this series, a mayor proportion of 44.68%of patients belong to the group between 16 to 35 years old. The group above 35 years accounts 30%.The age groups of 6 to 15 years old and the group of 1 to 5 year old account 16.48% and 8.51%, respectively.
Finally, it is important to point out that 37.76% of our hemophiliac patients are in the group of moderate illness status and 44.14% in the group of severe illness status. All the patients receive treatment with plasma-derived coagulation factor concentrates and in some cases recombinant therapy. Children receive prophylactic treatment.

Disclosures:
No relevant conflicts of interest to declare.
Retrospective Review of Life-Threatening Bleeds in Paediatric ITP.
Authors: Killen Maire-Clare , Reeves Marie , Grainger John D. 
Published: October 2019

Abstract
Abstract 4453
Immune thrombocytopenia (ITP) is an uncommon disorder of childhood. Most children recover quickly regardless of intervention, but severe bleeding develops in approximately 3%. Intracranial haemorrhage (ICH) is by far the most feared complication, occurring in 0.1-0.5%. Pharmacotherapy is available to transiently increase platelet counts, but introduction of British clinical guidelines in 1992 has resulted in a sharp decline in the number of children receiving such treatment.
The aim of this survey is to establish the current UK incidence of intracranial and other life-threatening haemorrhages in ITP, and whether or not children developing such complications have other risk factors for bleeding.
UK paediatricians were mailed a questionnaire asking to report the number of cases of ITP per year, and any cases of intracranial haemorrhages (ICH) in the previous ten years or other life-threatening haemorrhages in the past five years. Consultants reporting patients were asked to complete a form requesting detailed information on the child and the circumstances surrounding their bleed.
844 annual cases of ITP were reported from 145 centres; 672 of these were acute presentations and 240 were chronic. 24 cases of ICH and 19 cases of life-threatening bleed were identified, giving an annual incidence of 0.3% and 0.5% respectively. Gastrointestinal haemorrhage was the most commonly reported life-threatening bleed, and in six cases, ICH and life-threatening bleeds occurred in the same child.
The incidence of ICH in ITP may be higher than previously reported in the UK; the reason behind this increase remains unknown and support the need for larger international collaborative studies.

Disclosures:
No relevant conflicts of interest to declare.
GRAFT-VS-LEUKEMIA EFFECT in a MOUSE MODEL of CHRONIC MYELOID LEUKEMIA.
Authors: Lamana María L , Oviedo Alberto , Yañez Rosa , Aldea Montserrat , Rubio Antonio , Bueren Juan A. 
Published: October 2019

Abstract
Abstract 4500
Graft-vs-Host Disease (GVHD) is a frequent and severe complication of allogeneic hematopoietic stem cell transplants (allo-HSCT), mediated by donor's T cells reacting against host antigens. However, donor's T lymphocytes also generate the beneficial Graft-vs-Leukemia effect (GVL) by recognizing tumor antigens as non-self, thus contributing to the eradication of residual leukemic cells. With the purpose of studying factors that could affect the GVHD/GVL effects, we generated a chronic myeloid leukemia (CML) mouse model by the transplantation of lin- bone marrow (BM) cells transduced with a retroviral vector carrying the BCR/ABL and tNGFR marker genes (p210-tNGFR RV) into syngeneic, lethally irradiated, B6D2F1 (H2b/d) mice. Transplanted mice developed chronic myeloid leukemia (CML), characterized by hemorrhagic lungs, hepatomegaly, splenomegaly and granulocytosis. Additionally, NGFR+ leukemic cells were detected in the peripheral blood and bone marrow. All mice died on days +18-20 after infusion of the transduced cells. The co-transplantation of B6D2F1 mice with syngeneic p210-tNGFR transduced lin- BM cells together with allogeneic BM cells from C57Bl/6 mice (H2b/b), determined a slower progression of the disease. In this case, transplanted mice died of leukemia on days +28-72 after the co-infusion.
To investigate in this CML mouse model the GVHD/GVL effect mediated by donor allogeneic T-lymphocytes, splenocytes from allogeneic C57Bl/6 mice were additionally infused together with the allogeneic BM cells and the symgeneic p210-tNGFR transduced lin- cells. No CML signs developed, and no NGFR+ cells were detected in mice receiving the allogeneic T cells. In this experimental group, however, all the animals died from acute GVHD on days +13-36 after the co-transplantation, similar to the GVHD control group that received allogeneic bone marrow cells and T-lymphocytes but not the p210-tNGFR transduced lin- cells.
In previous works, we demonstrated that the infusion of adipose tissue-derived mesenchymal stromal cells (Ad-MSCs) prevented GVHD in a haploidentical hematopoietic progenitor cells transplantation mouse model (Yañez et al, Stem Cells 2006). With the present GVHD/GVL mouse model we are now investigating the impact of the infusion of the immunosuppressive adipose tissue-derived mesenchymal stromal cells on the GVHD/GVL effect mediated by allogeneic T cells.

Disclosures:
No relevant conflicts of interest to declare.