bluuebunny/crossref_metadata_2025_split huggingface dataset:
The PI3K Inhibitor GDC-0941 Attenuates Disease in a KrasG12D Mouse Model of CMML and JMML.
Authors: Akutagawa Jon , Dail Monique , Friedman Lori S , Shannon Kevin M. , Sampath Deepak , Braun Benjamin S. 
Published: October 2019

Abstract
Abstract 2862
Chronic and juvenile myelomonocytic leukemias (CMML and JMML) are overlap myelodysplastic/myeloproliferative neoplasia (MDS/MPN) syndromes that respond poorly to conventional treatment regimens. Both diseases are characterized by aberrant N-Ras, K-Ras, Cbl, and SHP-2 proteins, which are not attractive drug target candidates. Focus has shifted to downstream effector pathways, which include Raf/MEK/ERK, phosphoinositide-3-OH kinase (PI3K)/Akt, and Ral-GDS/Ral-A cascades. However, it is unclear which pathways, if any, should be targeted. In part to address this question, we previously developed a mouse model of CMML and JMML by expressing a conditional “knock-in” KrasG12D oncogene in bone marrow. Our earlier studies showed that inhibition of MEK yields a significant reduction in disease burden in this model, including reduced leukocytosis, improved anemia and enhanced survival. Here, we interrogate the role of the PI3K/Akt pathway in KrasG12D driven MPN and further explore which specific pathways are responsible for leukemogenesis due to hyperactive Ras. We administered GDC-0941, a selective PI3K inhibitor, to Mx1-Cre, KrasG12D mutant mice Mice with well established MPN and wild-type (WT) littermates were randomly chosen to receive daily oral administration of GDC-0941 or a control vehicle. Treated mice exhibited dramatic corrections of leukocytosis and anemia as well as decrease in splenomegaly. Flow cytometry of bone marrow and peripheral populations also imply that GDC-0941 treatment corrects the aberrant proliferation, amends differentiation of bone marrow progenitors, and revives ineffective erythropoiesis found in KrasG12D mice. Treatment also resulted in markedly improved survival of KrasG12D mice; virtually all KrasG12Dmice in the treatment arm outlived their control counterparts. Our data suggest PI3K inhibition may play a role in suppressing hematologic dysfunction in JMML and CMML patients. Potential crosstalk between PI3K and MEK signaling further suggest that combinatorial activities of PI3K and MEK inhibition should be investigated.

Disclosures:
Friedman: Genentech, Inc.: Employment. Sampath:Genentech, Inc.: Employment.
Gene Expression Profiling (GEP) in MGUS and AMM: Predictors of Progression.
Authors: Waheed Sarah , Heuck Christoph , Qu Pingping , van Rhee Frits , Usmani Saad Z , Zhang Qing , Sexton Rachael , Crowley John , Epstein Joshua , Dhodapkar Madhav V. , Shaughnessy John D , Barlogie Bart 
Published: October 2019

Abstract
Abstract 2933
Monoclonal gammopathy of undetermined significance (MGUS), a premalignant plasma cell dyscrasia, is differentiated from asymptomatic multiple myeloma (AMM), based on the level of monoclonal immunoglobulin (M spike) and bone marrow plasma cell infiltration (M spike > 3 g/dl and/or marrow plasmacytosis >10% in AMM). MGUS progresses to active Multiple Myeloma (MM) at a rate of 1–2% per year, thus imparting an average risk of 25% of progression over a lifetime. No single clinical variable, imaging modality or molecular study has yet been identified to predict progression. As a part of SWOG observational trial S0120, 262 patients were enrolled at MIRT and their GEP of purified plasma cells (PC) prospectively obtained. Here we report the results of GEP as predictor of progression. Among those with baseline GEP data available, 127 were non-IgM, AMM or MGUS and were otherwise eligible for the study. Baseline samples were restricted to those samples taken no more than 6 months prior to or 14 days following registration to S0120. If a patient had multiple baseline samples, then the sample taken closest to the patient's registration date was used. GEP studies were performed to assign molecular classification and risk scores, along with GEP-derived amp1q21, delTP53, DKK1 and other variables. With a follow-up of 36 months in the subset with evaluable GEP baseline data in 40 MGUS and 87 AMM cases, GEP characteristics were compared with those of untreated MM patients treated with TT2, TT3, TT4, and TT5 protocols, in the context of normal donor PC signatures. Comparing MGUS with AMM, CD-2 molecular subgroup was over-represented in MGUS and HY in AMM. PSMD4 on chromosome 1q21 was also linked to AMM. In terms of GEP-70 risk model, 36% of AMM and 15% of MGUS had scores exceeding -0.26. Univariate predictors for treatment-requiring MM included the well-known serum-M and bone marrow PC variables, and GEP-70 score >-0.26 (HR=8.82, p<0.001). Whole bone marrow biopsy (BMB) GEP samples from MGUS/AMM were available in 26 samples. Applying a BMB-65 model distinguishing MM-BMB from NL-BMB (normal donors) and MGUS/AMM-BMB, we observed that there was a strong trend for delayed MM progression when MGUS/AMM-BMB scores were in the top tertile (most NL-like). Independent validation data will be presented.

Disclosures:
Dhodapkar: Celgene: Research Funding; KHK: Research Funding.
Granulocyte Colony-Stimulating Factor-Mediated Immunoregulation Via Inducing Blood Cells Expressing Human Leucocyte Antigen-G.
Authors: Liu Hui , Du Xuan , Chen Yinkui , Xuan Li , Wu Xiuli , Zhang Yu , Fan Zhiping , Huang Fen , Zhang Xian , Zhai Xiao , Zhang Fuhua , Sun Jing , Liu Qifa 
Published: October 2019

Abstract
Abstract 3051

Background
Peripheral blood stem cells (PBSCs) obtained from granulocyte colony-stimulating factor (G-CSF)-mobilized donors have been used more frequently than bone marrow stem cells as the source of cells in allogeneic hematopoietic stem cell transplantation (allo-HSCT). Although G-CSF–mobilized PBSC grafts contain more mature T cells than bone marrow cell grafts, the incidence and severity of graft-versus-host disease (GVHD) were not elevated. G-CSF-induced immune tolerance might be via inducing T helper type 2 (Th2) polarization, the promotion of regulatory T cell and tolerogenic dendritic cell (DC) differentiation. However, these mechanisms are not fully understood. Human leucocyte antigen-G (HLA-G) is a tolerogenic molecule which participates in the regulation of immune response. In this study, to explore the mechanisms of G-CSF-induced immunoregulation, the effect of G-CSF on blood cells expressing HLA-G was studied.

Methods
Membrane-bound HLA-G (mHLA-G) was detected using flow cytometry analysis; soluble HLA-G (sHLA-G), interferon gamma (IFN-γ) and interleukin 10 (IL-10) were determined by enzyme-linked immunosorbent assay (ELISA); HLA-Gpos cells were isolated using flow cytometry, and mixed leukocytes reaction (MLR) was carried out to assess the suppressive effect of HLA-Gpos cells.

Results
CD3+CD4+ T cells, CD3+CD8+ T cells, CD19+ cells and CD14+ cells all expressed mHLA-G in peripheral blood (PB) and bone marrow (BM) from 18 healthy donors before and after G-CSF mobilization. The levels of sHLA-G and mHLA-G on these cells in PB and BM all increased significantly after G-CSF mobilization (all P<0.05). The levels of sHLA-G and mHLA-G on these cells in BM were all higher than that in PB, including before and after G-CSF mobilization (all P<0.05). Bone marrow mononuclear cells (BMMCs) were stimulated with G-CSF in vitro, and the levels of mHLA-G on these cells and sHLA-G in culture supernatant all increased significantly after BMMCs cultivated with G-CSF for 24 hours (all P<0.001). In addition, the levels of IFN-γ and IL-10 also elevated in culture supernatant (both P<0.05). Antibodies blocking experiments for IL-10 and IFN-γ showed that IL-10 and IFN-γ were not necessary for G-CSF-induced HLA-G expression of blood cells. The results of MLR showed that HLA-Gpos cells were able to suppress the proliferation of allogeneic lymphocytes.

Conclusion
G-CSF could directly induce blood cells expressing HLA-G, which might be another mechanism of G-CSF-mediated immunoregulation in G-CSF–mobilized PBSC transplants.

Disclosures:
No relevant conflicts of interest to declare.
Airway Dilation in Bronchiolitis Obliterans After Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Gazourian Lee , Coronata Anna Maria F , Rogers Angela J , Weinhouse Gerald L , Soiffer Robert J. , Antin Joseph H. , Ritz Jerome , Ho Vincent T , Baron Rebecca M , Washko George R 
Published: October 2019

Abstract
Abstract 3058

Rationale:
Bronchiolitis obliterans syndrome (BOS) is a late, non-infectious pulmonary complication following hematopoietic stem cell transplantation (HSCT). The prevalence is estimated to be between 5–10% after allogeneic HSCT with a significant increase in morbidity and mortality. We were interested in assessing quantitative radiologic characterization of airway remodeling in these subjects.

Objectives:
To examine quantitative measurements of airway morphology and their correlation with lung function in a cohort of patients who underwent HSCT and developed BOS.

Methods:
All patients who underwent an allogeneic HSCT at the Dana-Farber Cancer Institute/Brigham and Women's Hospital (n=1854) between January 1st 2000 and June 30th2010 were screened for the development of BOS using modified NIH diagnostic criteria. Clinically acquired high resolution CT (HRCT) scans of the chest were collected. Discrete measures of airway wall area were performed in a total of 16 manually selected airways in the right and left upper and lower lobes using Airway Inspector (www.airwayinspector.org). From these measures, the square root of wall area of a 10-mm luminal perimeter (Pi10) was calculated. Spearman correlation was used to assess the relationship between square root of wall area at Pi10 and pulmonary function measures. Paired t-test was used for within group comparisons. Analyses were performed with SAS v. 9.2.

Measurements and Main Results:
Within our cohort 89 cases of BOS were identified of which 37 patients had HRCT scans available for analysis. 33 patients had pulmonary function tests within 30 days of the HRCT (Table 1). Ten of the 37 BOS patients had a HRCT scan at or prior to transplantation and a HRCT scan available after the diagnosis of BOS. Square root of wall area at Pi10 correlated significantly with FEV1(r = 0.636, p<0.0001), RV/TLC (r = −0.736, p< 0.0001) (Fig. 1), and when adjusted for age, sex and total lung capacity, the correlations remained significant (p<0.0001). There was a significant decline in square root of wall area at Pi10 in BOS cases compared to pre-transplant CT scans (p<0.001) (Fig 2).

Conclusions:
On HRCT scan BOS is characterized by central airway dilation, the degree of which is correlated to decrements in lung function. Future studies will examine whether chest CT assessment of central airway dilation can be used as a sensitive and specific tool for diagnosis and staging of BOS.

Disclosures:
No relevant conflicts of interest to declare.
Influence of Micrornas Expression in Graft-Versus-Host Disease After Allogeneic Transplantation.
Authors: Koldehoff Michael , Maniera Christina , Steckel Nina , Elmaagacli Ahmet H. , Beelen Dietrich W. 
Published: October 2019

Abstract
Abstract 3062

Background:
MicroRNAs (miRNAs) are a family of 19–24 nucleotide noncoding RNAs, which affect the regulation of gene expression in eukaryotic cells by binding to a 3′-untranslated region which target messenger RNAs. MiRNAs play important roles in many cellular processes such as development, stem cell division, apoptosis and cancer. We profiled miRNAs expressed on patients who underwent allogeneic hematopoietic stem cell transplantation (HSCT).

Methods:
Here we analyzed in a retrospective study 50 patients (pts) for miRNAs expression (miRNA-21; -128; -146; -155; and -181) in whole blood that underwent allogeneic HSCT and analyzed their outcome. MiRNA expressions were performed by miRNA-specific real time RT-PCR.

Results:
In this cohort 15 pts received grafts from HLA-identical siblings (30%), 25 pts from matched (50%) and 10 pts from mismatched (20%) unrelated donors. Transplantat consisted of unmanipulated peripheral blood stem cells (n=41, 82%) or bone marrow (n=9, 18%). Of all pts, 12 (24%) had relapsed and 3 (6%) died of February 2012. There was no significant correlation between miRNA expressions from siblings and unrelated donors. For miRNA-21, we found a moderate down-regulation in mismatched unrelated donors (p<0.08) versus matched donors. Analysis of each miRNAs for relapse or TRM showed no statistically differences. Among these pts, 45 (90%) developed acute GVHD (18 pts had an acute GvHD of grade ≥2). There was a trend correlation between low level of miRNA-181 (327 ± 117% vs. 537 ± 575%; p<0.07) and miRNA-128 (158 ± 92% vs. 297 ± 485%, p<0.06) of pts with acute GvHD ≥ 2. In pts with severe acute GVHD (grade >3) comparing all other pts, we found significant reductions of miRNA-146 (355% vs. 1526%, p<0.0001), miRNA-21 (904% vs. 5526%, p<0.001) and miRNA-155 (5.66% vs. 3.52, p<0.04). However, no statistic differences were found in the miRNA expressions in regards to chronic GVHD.

Conclusions:
These results suggest that pts with low expression of miRNA-146, miRNA-21 and miRNA-155 confirms a relevant association of the development of acute GVHD and miRNA profilings could be an early indicator of severe acute GvHD.

Disclosures:
No relevant conflicts of interest to declare.
Short Term Treatment With Thrombopoietin-Receptor Agonists For Patients With Immune Thrombocytopenic Purpura (ITP) Before Splenectomy
Authors: Kalish Yosef , Spectre Galia , Varon David 
Published: October 2019

Abstract
The thrombopoietin-receptor agonists (romiplostim and eltrombopag) were approved recently as treatments for patients with immune thrombocytopenic purpura (ITP). Splenectomy remains a common second line treatment for ITP with the highest remission rate compared with alternative therapies. It has been reported that splenectomy can be safely performed in patients with a platelet count of 40-50,000 per cubic millimeter but many patients do not reach these values. Intravenous immune globulins (IVIG) are often used before splenectomy in order to increase the platelet count before surgery.
The aim of this study was to determine whether a short course of thrombopoietin-receptor agonists can be used as a reliable and safe treatment to increase the platelet count in patients with ITP before splenectomy.
Between 2010 and 2012, fifteen patients with ITP, all refractory to steroids, were scheduled for splenectomy. Treatment with thrombopoietin-receptor agonists (romiplostim or eltrombopag) was started 3 weeks before splenectomy. Eight patients received eltrombopag at a dose of 50 mg/day orally until 3 days before splenectomy. For romiplostim, a subcutaneous injection of 3 mcg/kg was given weekly to 7 patients. The last injection was given one week before splenectomy. Complete blood count was repeated every week and the dose of romiplostim was adjusted (up to 10 mcg/kg or down to 1 mcg/kg) based on the platelet count increment. Response was defined as a platelet count of 50,000 or more per cubic millimeter.
Mean platelet count before treatment was 11,000±8,000 cells per cubic millimeter. All patients, except one patient on romiplostim, responded to the treatment with a mean platelet count of 74± 25 cells per cubic millimeter on the day of splenectomy (p<0.01). Similar effect was noticed among responders of the two drugs. Four patients from the romiplostim group responded to a 3 mcg/kg dose. Two patients responded to increased doses of 7 and 10 and 10 mcg/kg. One patient did not respond to 10 mcg/kg of romiplostim but later responded to IVIG. The two drugs were well tolerated with no side effects except for mild liver function abnormalities in one patient in the eltrombopag group. No thromboembolic complications or excessive bleeding were reported for these patients.
In summary, we report that a short course of thrombopoietin-receptors agonists can effectively and safely increase the platelet count in steroid resistant ITP patients before splenectomy.

Disclosures:
No relevant conflicts of interest to declare.
Immune-Mediated Activation Of Coagulation In Patients With Stevens Johnson Syndrome/Toxic Epidermal Necrolysis
Authors: Iqbal Omer , Syed Daneyal , Mosier Michael , Malalis Julia , Mata Christine , Lin Amy , Mitchel Nicole , Abro Schurahazard , Hoppensteadt Debra , Fareed Jawed , Bouchard Charles 
Published: October 2019

Abstract

Background
Stevens Johnson Syndrome/Toxic Epidermal Necrolysis (SJS/TEN) are specific drug hypersensitivity reactions initiated by cytotoxic T-lymphocytes. Augmentation of T-cell activation by positive co-stimulation and Major Histocompatibility Complex-restricted presentation of the culprit drug triggers T-cell activation following interaction with T-cell receptors. This T-cell activation results in expression of cytokines including TNF-α, interferon-γ, granzyme B and granulysin from NK cells. Besides their role in causing SJS/TEN, these cytokines cause coagulation activation.

Materials & Methods
Following an Institutional Review Board approved protocol blood samples were obtained from subjects suspected of SJS/TEN and normal healthy volunteers (n=2). Confirmatory biopsies were performed in all SJS/TEN subjects which confirmed SJS/TEN in 4 subjects and were negative in 7 subjects. Cytokine levels were measured using the cerebral II array biochip from Randox Laboratories Limited (Crumlin, UK). Besides cerebral array II, other parameters such as thrombin-antithrombin (TAT) complexes (Dade®, Marburg, Germany), fibrinopeptide A (F1.2, Dade®), plasminogen activator inhibitor-1 (PAI-1, Diagnostica Stago® and STACHROME antithrombin (Stago®), ZYMUPHEN platelet microparticles activity (Hyphen® Biomed (Neuville-sur-Oise, France), and HEMOCLOT protein C (Stago®) were measured using ELISA kits as per manufacturers’ instructions.

Results
Compared to the Normal Human Plasma, the IL-4, IL-6, TNF-α, and MCP-1 levels were increased significantly in biopsy-confirmed SJS/TEN patients (see table). Other parameters measured including IL-2, IL-8, IL-10, VEGF, INF-γ, IL-1α, IL1-β and EGF were not significantly increased. A marked increase in the TAT complexes (6.3±5.9 µg/ml), F1.2 (430.4±202.4 pmol/L), platelet microparticles (13.1±9.3 nM) and protein C levels (90.5±63.4%) with a corresponding decrease in PAI-1 (53.3±18.8ng/ml) and antithrombin levels (80.7±42.4%) compared to normal human plasma were also observed. Biopsy-negative SJS/TEN subjects with less pronounced inflammatory stimulus, demonstrated mild elevation in cytokine levels of IL-4 (1.95±0.59 pg/ml), IL-6 (29.81±25.18 pg/ml), TNF-α (7.20±5.04 pg/ml) and MCP-1 (265.10±159.09 pg/ml)).

Conclusions
T-cell activation and release of cytokines especially TNF-α and granzyme B causes immune-mediated activation of coagulation with increase in TAT, MCP-1, F1.2 and platelet microparticles and corresponding decrease of protein C, antithrombin, and PAI-1. These alterations in coagulation may progress to sepsis associated coagulopathy and overt disseminated intravascular coagulation.

Disclosures:
No relevant conflicts of interest to declare.
Clinical Study Of DC-CIK (dendritic cells and cytokine-induced killer cells ) Eliminate Minimal Residual Leukemia
Authors: Zheng Zhuanzhen , Yuna Zhang , Huimin Zhang , Liu Qingchi , Ma Yanping , Wu Weihai , MA Chuanbao , Pang Yuhui , LI Jianying 
Published: October 2019

Abstract

Objective
To evaluate the clinical efficacy and safety of allogenic dendritic cells (DCs) and cytokine-induced killer (CIK) cells in the eliminating minimal residual leukemia (MRL).

Methods
48 acute leukemia patients with hematological complete remission (CR) but without molecular biological remission (CRM), or patients with minimal residual Leukemia (MRL) were selected from Ping’an Hospital of Shijiazhuang during Jan. 2009 to Jun. 2011. According to the patients’ will, 48 patients divided into combined treatment group and chemotherapy group 24 each. All the patients were in the same general information and disease level. The combined treatment group was treated with DC-CIK and consolidation chemotherapy, and the chemotherapy group was treated with consolidation chemotherapy. PBMCs were collected from healthy donors (the patient's parents or children) to prepare DC-CIK cells. DC-CIK cells were intravenous injected into patients once every 15 days, a total of 4-6 times infusion. The blood routine, bone marrow cells, leukemia related genes, urine and stool routine, liver and kidney biochemistry function, and ECG were observed. Changes of peripheral lymphocyte subsets in patients were detected by flow cytometry. Adverse reactions were examined.

Results
(1)Eleven cases in the combined treatment group achieved CRM, and the CRM rate was 45.8%; whereas only 2 cases in the chemotherapy group achieved CRM and the CRM rate was 8.3%,the difference was statistically significant(χ2=8.55, P<0.01).(2) Compared with the chemotherapy group, the CFIM (four-color combination flow cytometric immunophenotype of minimal residual leukemia) negative conversion rate of patients in the combined treatment group was significantly raised (66.7% vs 25.0%,χ2=8.39, P<0.01). (3)The negative conversion rate of MRL was higher in the combined treatment group than the chemotherapy group (66.7% vs25.0%, χ2=8.39, P<0.01). (4) After treatment the ratio of CD4+/CD8+ cells was significant increased than before treatment in the combined treatment group (1.3±0.4 vs 0.8±0.4, P<0.05). (5)The complete remission rate (CCR) of patients in the combined treatment group after 3 years was 79.2%, while that in the chemotherapy group was 45.8% (χ2=5.69, P<0.05).(6)No dysfunction of critical organs such as heart, liver and kidney and serious adverse reactions were observed while DC-CIK cells infusion.

Conclusion
DC-CIK combined with chemotherapy can inhibit leukemia gene, promote the negative conversion rate of CFIM, facilitate the clear of MRL, improve immune function and prolong remission of the patients.No serious adverse reactions were found in patients with DC-CIK infusion.

Disclosures:
No relevant conflicts of interest to declare.
Puma Is The Critical BH3-Only Protein Mediating Apoptosis In The Nup98-HoxD13 (NHD13) Mouse Model Of Human MDS
Authors: Guirguis Andrew , Curtis David J. , Slape Christopher , Saw Jesslyn , Wei Andrew H , Failla Laura M , Bouillet Philippe , Strasser Andreas 
Published: October 2019

Abstract
One of the key hallmarks of myelodysplastic syndromes (MDS) is that of ineffective haematopoiesis. The main factor contributing to this is increased apoptosis of bone marrow progenitors – a prominent feature particularly of early-stage MDS. Although apoptosis has been attributed to cell extrinsic triggers such as increased levels of Fas and TNF-α, there is increasing evidence implicating the cell intrinsic pathway – in which Bcl-2 and its related proteins play an important role and which is activated by p53. Consistent with this, we have recently shown that apoptosis in the Nup98-HoxD13 (NHD13) transgenic mouse model of MDS can be effectively blocked by enforced expression of Bcl-2. To further define the molecular mechanism of apoptosis upstream of Bcl-2, we mated NHD13 mice with mice deficient for p53. Consistent with a lineage specific role for p53, we observed p53-dependent apoptosis of the erythroid, but not the myeloid lineage. In NHD13 progenitors, gene expression analysis for members of the intrinsic pathway, showed elevated expression of Noxa but not Puma – both of which are BH3-only proteins and transcriptional targets of p53. We proceeded to mate NHD13 mice with mice deficient in Noxa and Puma. Unexpectedly, the expression levels were in fact misleading as genetic ablation studies showed that apoptosis of both erythroid and myeloid lineages was Puma dependent, with no role for Noxa. These studies have demonstrated that gene expression of Bcl-2 proteins cannot be used to predict functional relevance and in this case, Puma is the critical BH3-only protein mediating apoptosis in MDS.

Disclosures:
Guirguis: Leukaemia Foundation - Australia: Research Funding.