bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genomic Analysis of Serial Chronic Lymphocytic Leukemia Samples Suggests That Epigenetic Changes, Rather Than Clonal Evolution, May Drive the Progression of This Leukemia
Authors: Ghia Emanuela M. , Smith Erin , DeBoever Christopher , Rassenti Laura Z. , Rozenzhak Sophie , Shepard Peter , Alakus Hakan , Harismendy Olivier , Barrett Christian L , Frazer Kelly A , Kipps Thomas J. 
Published: October 2019

Abstract
Abstract 4563
The clinical course of patients with chronic lymphocytic leukemia (CLL) is heterogeneous, with some patients requiring treatment relatively soon after diagnosis and others having indolent disease for many years. Some patients with indolent disease, however may develop more aggressive disease over time that requires therapy. To identify genetic and epigenetic changes that associate with the transition from indolent to aggressive disease, we used genomic methods to analyze sequential samples obtained from 19 CLL patients evaluated at the UC San Diego Moores Cancer Center who ultimately required treatment, as per iwCLL guidelines.
For all patients, the first time point sample collection (SC1) was obtained within 1 year post-diagnosis and the second time point sample collection (SC2) was obtained within 1 year before treatment. We performed whole-exome sequencing (Agilent 50Mb capture, 100×) and methylation (450K) array analyses on leukemia cells and germline DNA. Somatic allele frequencies ranged from < 10% to 50%, suggesting heterogeneity within the tumor. When comparing SC1 versus SC2, we observed changes in somatic allele frequency for 6 (32%) of 19 patients, however 13 (68%) of 19 patients did not have evidence for clonal evolution at the somatic level, suggesting that the acquisition of additional somatic mutations did not drive CLL progression and that the clonal population structure remains stable throughout disease progression with multiple clones per patient. Using 450K CpG methylation arrays, we identified 52,409 sites (FDR=0.05) that changed consistently between SC1 and SC2 across 19 patients, suggesting that epigenetic changes were widespread, even without detectable somatic mutations.
In summary, our results imply that CLL progression can occur in the absence of somatic mutations, but rather may reflect non-stochastic alterations in the epigenome altering RNA expression.

Disclosures:
No relevant conflicts of interest to declare.
Divergence in CD19-Mediated Signaling Unfolds Intra-Clonal Diversity in Chronic Lymphocytic Leukemia Which Correlates with Disease Progression
Authors: Herishanu Yair , Kay Sigi , Dezorella Nili , Perry Chava , Deutsch Varda , Polliack Aaron , Naparstek Elizabeth , Katz Benzion 
Published: October 2019

Abstract
Abstract 4570
Emerging data on intra-clonal diversity imply that this phenomenon may play a role in the clinical outcome of patients with chronic lymphocytic leukemia (CLL), where subsets of the CLL clone responding more robustly to external stimuli may gain a growth and survival advantage. Here we report intra-clonal diversity resolved by responses to CD19 engagement in CLL cells, which can be classified into responding (CD19-R) and non-responding (CD19-N) sub-populations. Engagement of CD19 by anti-CD19 antibody rapidly induced cellular aggregation in the CD19-R CLL cells. The CD19-R CLL cells expressed higher surface levels of CD19 and c-myc mRNA and exhibited distinct morphological features. Both sub-populations reacted to IgM stimulation in a similar manner and exhibited similar levels of Akt phosphorylation, pointing to functional signaling divergence within the B-cell receptor. CD19 unresponsiveness was partially reversible, where CD19-N cells spontaneously recover their signaling capacity following incubation in vitro, pointing to possible in vivo CD19-signaling attenuating mechanisms. This concept was supported by the lower CD19-R occurrence in bone marrow-derived samples compared with cells derived from the peripheral blood of the same patients. CLL patients with more than 18.5% of CD19-R cell fraction had a shorter median time to treatment compared to patients with less than 18.5% of CD19-R cell fraction.

Conclusions:
Divergence in CD19-mediated signaling unfolds both inter-patient and intra-clonal diversity in CLL. This signaling diversity is associated with physiological implications including the location of the cells and disease progression.

Disclosures:
No relevant conflicts of interest to declare.
Array Comparative Genomic Hybridization: A Better Alternative for Genomic Profiling and Prognosis of Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma
Authors: Leon Annette , Sevilla Deborah , Marino Joseph M , Moreno Kenneth D , Obrera Rev , Lee PoShing , Ye Donjiu , Talwar Jotica , Persad Rajendra , Villamil Cesar , Maya Melissa , Mehta Nirav , Kemel Yelena , Weisberger James 
Published: October 2019

Abstract
Abstract 4572
Traditional cytogenetic techniques such as chromosome analysis and fluorescence in situ hybridization (FISH) have provided valuable genetic information in the evaluation of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL). However, many genomic abnormalities remain undetected due to the limitations intrinsic to these techniques. We designed, validated, and clinically applied a combined targeted-whole genome custom oligonucleotide microarray for the evaluation of hematological malignancies. Incorporation of array comparative genomic hybridization (aCGH) analysis has enabled us to identify previously undetected copy number changes with an increased resolution and sensitivity and more precisely determine genomic breakpoints and gene content with diagnostic and prognostic relevance in CLL/SLL. A cohort of 300 patients diagnosed with CLL/SLL was studied using concurrent cytogenetics, FISH and aCGH analyses on either peripheral blood or bone marrow aspirate samples. Array CGH identified clinically significant genomic alterations in 75% of the cases compared to 57% and 47% by cytogenetics and FISH analyses, respectively. There were no statistically significant differences detected between peripheral blood and bone marrow samples. Recurrent alterations such as deletions in chromosomes 11q (ATM, adverse prognosis), 13q (RB1, DLEU1, 2, and 7, MIR15A and 16–1, favorable prognosis in general), 17p (TP53, adverse prognosis) and gain of one copy of the entire chromosome 12 (ETV6, KLRK1, CD27, KRAS, MDM2, HIP1R, KITLG, intermediate prognosis) were identified in 55% of cases by aCGH. Of these cases, chromosome and/or FISH analyses did not detect these abnormalities in 42% and 27% of cases respectively. In addition, aCGH study more fully characterized the interstitial deletion in chromosome 13q recently suggested to have different outcomes based on size and gene content. Important findings exclusively by aCGH analysis in 7% of cases include loss of genetic material in chromosomes 3p (KAT2B, FHIT, clonal evolution), 6q (MAP3K7, PRDM1, AIM1, TNFAIP3, intermediate prognosis), 8p (TNFRSF10D, disease progression), 9q (JAK2, MLLT3, CDKN2A and B, DAPK1) and gain of genetic material in chromosome 2p (ACP1, MYCN, ALK, REL, BCL11A) associated with unmutated IGHV status, advanced disease stage and adverse prognosis. The results obtained in this study demonstrate the superior resolution and detection rate of aCGH technology as well as the importance of incorporating this methodology into current algorithms for the diagnosis and prognosis of CLL/SLL.

Disclosures:
No relevant conflicts of interest to declare.
Forest Plot to Compare Therapies in a Network Meta-Analysis in First Line Chronic Lymphocytic Leukemia
Authors: Piciocchi Alfonso , Bontempi Katia , Fazi Paola , Vignetti Marco , Mandelli Franco 
Published: October 2019

Abstract
Abstract 4596
Several therapies, composed by single agent or combinations, are available for untreated patients in chronic lymphocytic leukemia and it is difficult to rank treatments according to their effect size when not all treatments have been compared directly. Direct randomized comparison is the most reliable way of comparing treatments but when the number of available treatments increases the number of possible pairwise comparisons increases quadratically, so it is common for only a small fraction of the possible comparisons to be performed. Network meta-analysis permits to add indirect estimates for assessing the relative effectiveness of two treatments when they have not been compared directly in a randomized trial but have each been compared to other treatments. Network meta-analysis of randomized controlled trials was used to combine direct and indirect estimates of the effect of four drugs and their combinations from five randomized controlled trials (Fig. IA) in CLL patients based on a systematic review conducted by M.M. Cheng et al. (Cancer Treatment Review 2012) on 2625 patients where Chorambucil, Fludarabine, Rituximab, Alemtuzumab, Bendamustine, Cladribine and Pentostatin have been considered as primary first-line agents for CLL. Patients were younger (59–65 years), had good performance status (ECOG: 0–1) and early stage disease (Rai 0-II, Binet A or B). We carried out an analysis based on the endpoint of PFS, described as the duration of the time from randomization until disease progression or death from any cause and to consider the heterogeneity among the studies a random effects model was performed. The differences in PFS of all therapies considered in the analysis were compared with combination of Fludarabine and Chorambucil (FC). The results were summarized by forest plot (Fig IB) in terms of reduction or increase of Hazard Ratio. As the forest plot shows, FCR has relatively higher potential of preventing disease progression in younger patients affected by chronic lymphocytic leukemia.

Respect to a traditional meta-analysis, which usually compares only two treatments, network meta-analysis is a useful method to combine direct and indirect comparisons of treatments from RCTs and to analyze the hierarchy in treatment effects and tests for consistency of the relations of the network.

Disclosures:
No relevant conflicts of interest to declare.
A Neonate with Type 3 Von Willebrands Disease and Life Threatening Bleeding Treated Successfully with a FVIII/VWF Concentrate (wilate®)
Authors: Hegener Oliver , Efford Jemma , Bartram Jack L , Henderson Lesley , Khair Kate 
Published: October 2019

Abstract
Abstract 4633

Case presentation
A 1 day old male presented to local hospital in collapsed state following normal vaginal delivery at 41 weeks gestation. The baby had sudden onset of limpness at home. On arrival at hospital he was making no spontaneous movement had slowly reactive pupils, oxygen saturation was 76% in air. He was intubated and blood was noted on his vocal cords. His haemoglobin was 2.3g/dl and coagulation grossly abnormal with APTT of >100seconds. There was no family history of bleeding diathesis. The baby was transferred to neonatal intensive care unit at this hospital.

Management
On arrival at this hospital the baby was ventilated, had received both FFP and blood transfusion and an additional dose of Vitamin K whilst clotting factor assays were awaited. Abdominal distention was noted and an ultra sound of the abdomen revealed a large collection of blood in the abdominal cavity. Severe vWD (FVIII 8iu/dl and VWF Rapid 5iu/dl post FFP) was diagnosed. Treatment with Wilate 60iu/kg/dose was initiated and continued for 7 days during which time there was no further bleeding.

Outcomes
The baby was ventilated for 72 hours, a central line was inserted for venous access and administration of Wilate. Subsequent factor assays (with out FFP) reveal FVIII <1 iu/dl and vWF <1 iu/dl. The baby was discharged from hospital aged 2 weeks and has had no further major bleeds but has been treated with Wilate for minor bleeding following trauma. He is now 7 months old and is thriving and well. Genetic analysis confirms severe vWD inherited in an autosomal recessive manner.

Conclusion
The FVIII/VWF concentrat, Wilate, proved effective in achieving lasting haemostatsis in the management of this acutely unwell neonate with significant spontaneous intra-abdominal bleeding.

Disclosures:
Hegener: Octapharma: Employment.
Regulatory Role of Host IL-17 Via Control of Host Macrophage Activation Contributes to Less Acute Gvhd
Authors: Kuroda Rie , Sato Katsuaki , Maeba Hideaki , Fujiki Toshihiro , Mase Shintaro , Araki Raita , Naka Kazuhito , Naito Makoto , Iwakura Yoichiro , Yachie Akihiro , Koizumi Shoichi , Nishimura Ryosei 
Published: October 2019

Abstract
Abstract 4669
IL-17 has emerged as a key pro-inflammatory cytokine, however, recent studies demonstrated that IL-17 has also anti-inflammatory effects. In addition, a wide variety of IL-17 producing cells are identified not only in T-cells but also in subpopulation of monocytes and macrophages, mast cells, Paneth cell in the gut so on. Therefore, IL-17 mediated immune responses are becoming much more complicated. In hematopoietic stem cell transplantation, we have reported that host-derived IL-17 has a protective effect against acute graft-versus-host disease (GVHD) using rodent models, in contrast, donor-BM-derived IL-17 exacerbates chronic GVHD. In acute GVHD model, lethally irradiated IL-17 knockout (KO) recipient mice receiving allogeneic BM with low dose splenocytes developed more severe acute gut GVHD compared to wild type (WT) host mice and finally half of them died (p<0.05). Much higher number of infiltrated monocyte/macrophage cluster was observed in spleen and gut in IL-17 KO host mice. Furthermore, residual host-typed peritoneal macrophages in IL-17 KO host mice were highly activated with the expression of TNF-α, while activation of donor-typed macrophages was much less with the production of anti-inflammatory cytokine IL-10. It suggested that these activated and increased macrophages might deeply involve in the pathogenesis of lethal GVHD. To test this, IL-17 KO recipient mice were given lipoCL2MDP intraperitoneally on day -7 before BMT to deplete host macrophages. As shown in the figure below, the macrophages depleted IL-17 KO host mice given splenocytes to induce lethal GVHD showed a significant survival benefit compared with IL-17 KO mice without depletion of macrophages. In conclusion, host IL-17 has a protective effect against acute GVHD with suppression of the activated macrophages. This is the first report that IL-17 has the immuno-regulatory role via control of activated host macrophages in acute GVHD.

Disclosures:
No relevant conflicts of interest to declare.
Surveillance of Healthcare Associated Infection in the Adult Hematology Unit
Authors: Lamchahab Mouna , Cherkaoui Siham , Ouhaddouss Mohamed , Qachouh Meryem , Benchekroun Said , Quessar Asmaa 
Published: October 2019

Abstract
Abstract 4712

Background:
Patients with hematological malignancies are at risk for Healthcare Associated Infection (HAIs) due to malignancy types, antineoplastic chemotherapies, immunosuppressant treatment, and prolonged hospital day. HAIs cause delay in cancer treatment, increase morbidity or mortality and increase costs of treatment affecting successful outcomes of cancer care. We describe the outcome of the surveillance in adult hematology patients in our hospital and identified groups most at risk for HAIs. We describe the outcome of the surveillance study in adult hematology patients in Morocco. Methods: A prospective surveillance study included all patients admitted to the adult hematology unit from March 2011 to August 2011. They excluded patients from the transplant unit and day hospital. The Center for Disease Control and Prevention standard definitions were used to determine if an infection was hospital acquired. We collected data in demographics, risk factors, the presence or absence of HAIs and its type. Results: A total of 180 patients that represented 377 admissions, with 4438 hospitalization-days, were included in the study. They have a mean age of 35 years (18–78 years). The sex-ratio H/F was 1.7. The distribution of the patient's diagnosis is related in Table1. 63 patients had 110 episodes of HAIs. The incidence rate of HAIs was 24.74/1000 hospitalization-days. The most frequent of infectious episodes were bacteremia (38.18%) and pneumonia (24.54%). The fever of unknown origin represented 35.45%.The distribution of the intrinsic risk factors with regard to episodes of HAIs was the following: 83.48% presented with neutropenia, 59.79% presented with severe neutropenia. With regard to extrinsic risk factors for HAIs, 61.03% of the infectious episodes had central venous catheterization. The mortality rate due to HAIs was 8%. Conclusion: Although the overall HAI rate is in line with published reports in hematology units, our mortality rate is higher. This finding suggests that we must be more aggressive to prevent infections in the patients in these units. Table I: Distribution of patient's diagnosis and incidence rate of HAIs Infection Incidence rate (/1000 hospital day) N n % Age (years) 18–40 41–60 > 60 Hematologic neoplasia 245 97 35 70 31 9 28,57 31,96 25,71 Acute myeloid leukemia 147 98 89,00 35,27 Acute lymphoblastic leukemia 83 5 4,54 Non Hodgkin's lymphoma 102 5 4,54 Hodgkin's disease 36 0 Aplastic anemia 3 2 Others 6 0 Total 377 110 100

Disclosures:
No relevant conflicts of interest to declare.
Peripheral Blood Lymphocyte and Platelet STEM CELL Expansion in Autologous Platelet-Derived Growth Factor-RICH Plasma
Authors: Salas Rodolfo J , De Salvo Dalila , De Salvo Cardullo Luigi , Hassanhi Manzur , Paz Luis S , Salas Dalida , Fuenmayor Alba , Acurero Zulay , Carrizo Ricardo , Herrera Humberto , Nunez Iris 
Published: October 2019

Abstract
Abstract 4717
Research on stimulatory and inhibitory factors present in the hematopoietic microenvironment promoted development of ex-vivo cell culture methods. Recent investigations strive to select natural and novel mitogenic substances within defined conditions to culture and harvest specific progenitor cells and cell lines. Optimal large scale cell expansion of HSCs remains to be developed. The present study documents ex-vivo human lymphocytes and platelet culture in autologous human blood plasma (BP) complemented with a novel ionic metal polysulphide. Resulting plasma is growth-factor rich (FRP) with a high platelet count (HPC) containing > 4.5×10E6 plts/ul. Co-culture expansion for 3-days of normal peripheral blood cells from n=20 healthy individuals in FRP promotes lymphoid and platelet series stem cell lineage growth and differentiation. Cultured cells characterized using conventional hematology and flow cytometry methods on CD3, CD4, CD8, CD34, CD38, CD41, CD61 antigen bearing cells; and average values reported. Result analysis of CD3 and CD4 bearing cells before-after a 3-day culture indicates a 4 % and 18 % respective increase, while CD8 cell population decrease 7%. A stimulation response obtained on CD34 and CD38 cells with a 67.30% and 53.89 % respective increase. Cell growth of CD41 and CD61 antigen bearing cells increased 50.89 % and 69.22 %, respectively. Functional platelet counts (before-after culture) correspond with a 3–5 fold increase of original concentration, and presence of recently reported proto-platelets. Minimum and maximum values varied among studied population. Discrete elevation in Neutrophils and Granulocytes counts observed. Potential applications thought for immunosuppression, cancer and organ transplant.

Disclosures:
No relevant conflicts of interest to declare.
Eltrombopag, but Not Romiplostim or Thrombopoietin, Inhibits Growth of Thrombopoietin Receptor Positive and Negative Human Leukemia Cell Lines
Authors: Rusnak David W. , Rudolph Sharon K. , Safavi Afshin , Erickson-Miller Connie L. 
Published: October 2019

Abstract
Abstract 4726
The thrombopoietin receptor agonists (TPO-RA), romiplostim and eltrombopag, are presently indicated for the treatment of certain patient groups with immune thrombocytopenia purpura. In a clinical study with romiplostim in patients with low-/intermediate-1 risk myelodysplastic syndromes (MDS), cases of transient increases in blast cell counts were observed and cases of MDS disease progression to acute myeloid leukemia (AML) were reported.
In the present study, we evaluated the impact of romiplostim, eltrombopag, and recombinant human thrombopoietin (TPO) on the proliferation of 5 human AML and 1 TPO-dependent megakaryoblastic cell line. The cell lines evaluated include the TPO-dependent cell line, N2C TPO; the TPO-R positive AML lines, HEL92.1.7 and OCI-AML-3; and the TPO-R negative AML cell lines, HL60, THP-1, and NOMO-1. All cells were exposed to 11-point dose response curves of the 3 agents at concentrations sufficient to generate a full stimulatory response in the N2C TPO cell line. Cells were exposed to concentrations of romiplostim and eltrombopag that met or exceeded the reported Cmax achieved for each agent in high-dose clinical trials and were 3- (eltrombopag) to 30-fold (romiplostim) above trough levels from the same clinical trials.
Neither romiplostim nor TPO treatment resulted in detectable stimulation or inhibition of leukemia cell growth at concentrations up to 10 μg/mL. Treatment with eltrombopag up to 40 μg/mL caused inhibition of all AML cell lines with mean IC50 values ranging from 6.4 to 13.5 μg/mL. These IC50 values reflect concentrations that are 3- to 6-fold below the Cmax of a 300 qd dose of eltrombopag (40.5 μg/mL) and at concentrations as low as 2-fold below Ctau levels (12.4 μg/mL). Cmax exceeded the IC90 for these AML cell lines, which ranged from 18.5 to 27.9 μg/mL. No stimulation of AML growth was evident through the range of the eltrombopag dose response curve on any of the cell lines evaluated. The results of this study confirm earlier in vitro studies (Will 2009, Erickson-Miller 2010) showing inhibitory effects of eltrombopag on leukemic cell lines and support clinical studies to evaluate a potential anti-leukemic effect of higher doses of eltrombopag in patients with AML.

Disclosures:
Rusnak: GlaxoSmithKline: Consultancy, Equity Ownership, Patents & Royalties. Off Label Use: Eltrombopag is an oral TPO agonist indicated for chronic ITP being studied in MDS/AML. Rudolph:GlaxoSmithKline: Consultancy, Equity Ownership. Erickson-Miller:GlaxoSmithKline: Employment, Equity Ownership, Patents & Royalties, Research Funding.
New Method for in Vivo Hematopoiesis Microenvironment Research: Tissue-Engineering-Bone Femur Repairing Model and Semi-Solid Decalcification
Authors: Jiang Shan , Jiang Qianli , Zhu Liqiong , Chen Feili , Yuan Lezhong , Zhao Peiran , Yang Mo , Sun Jing , Meng Fanyi 
Published: October 2019

Abstract
Abstract 4732

Background:
Bone defection repairing is a good model to study the hematopoiesis and its environment, during which there are reformations of niches and marrow cavity, different cells' migrations and evolutions, etc. Tissue-engineering-bone (TEB) is a kind of scaffold that degrades gradually during bone reformation. In order to observe different cells growing within the rigid, opaque, friable and porous TEB, we established a novel technique named semi-solid decalcification (SSD).

Aim:
With the TEB repairing model, we hope to address 3 questions: Firstly, could a functional bone with marrow cavity be reformed with the TEB? Secondly, what are the contributions of cells within and outside the TEB? Thirdly, how do the hematopoietic cells cross-talking with the microenvironment during bone reformation?

Method:

Results:

Conclusion:
This eGFP-to-mRFP transgenic mice femur repairing model was proved, at least in principle, to be efficient to mimic the reforming of bone marrow and stem cell niche. With the help of SSD, we can easily study the relationship and cross-talking of cell-cell and cell-matrix in a 3-D pattern, which is of the most important to understand the mechanism of hematopoiesis microenvironment. However, further researches are carrying on, to exclude the possible mixture of eGFP+ hematopoietic cells into mesenchymal cells before TEB-MC-GFP implantation and to better identify the subtypes of existing cells within the TEB.
PS: Thanks to the support of NSFC (No.30901367, 31070866).

Disclosures:
No relevant conflicts of interest to declare.