bluuebunny/crossref_metadata_2025_split huggingface dataset:
Primary Immune Thrombocytopenia (ITP) Guidelines for Bone Marrow Aspirate Reduce the Diagnostic Rate of Myelodysplastic Syndromes
Authors: Vincelli Iolanda , Oliva Esther Natalie , Ronco Francesca , Cufari Patrizia , Oliva Bianca , Gatto Valentina , Laganà Carmelo , D'Errigo Maria Grazia , Nobile Francesco 
Published: October 2019

Abstract
Abstract 4974

Introduction:
Primary immune thrombocytopenia (ITP) is an acquired immune-mediated disorder characterized by isolated thrombocytopenia, defined as a peripheral blood PLT count <100 Gi/L, and the absence of any obvious initiating and/or underlying cause of the thrombocytopenia (Rodeghiero et al, Blood, 2009). Guidelines consider bone marrow (BM) aspirate informative in patients with thrombocytopenia > 60 years of age, those with systemic symptoms or abnormal signs or in some cases where splenectomy is considered (Provan et al, Blood, 2009).

Methods:
We performed a retrospective chart analysis between January 2008 and June 2010 of all patients referred to our clinic for isolated thrombocytopenia but with a PLT count 100– 149 Gi/L. According to recent guidelines, these patients are not to be considered thrombocytopenic and do not require further investigation. The aim of the study was to evaluate the validity of omitting BM analysis in these cases.

Results:
Twenty-three cases (13 males/10 females) of mean age 58 ± SD 19 years were evaluated at our clinic for a PLT count below normal lab range values. At the time of evaluation none had bleeding symptoms. PLT counts ranged from 101 to 149 Gi/L, mean 123 Gi/L. Four patients had an enlarged spleen. After initial screening, 2 patients had a complex autoimmune disorder and 1 case had HCV hepatitis. The remaining 20 patients had a bone marrow (BM) aspirate performed: a diagnosis of myelodysplastic syndrome (MDS, WHO classification refractory thrombocytopenia) was obtained in 13 cases (65%) and BM biopsy was performed in 12, completed by cytogenetics in 9 cases (7 normal, 1 del20q, 1 –Y). Patients diagnosed with MDS were significantly older (66 ± SD 13 vs 47 ± SD 21 years, p = 0.017), but 4 cases (31%) were < 60 years of age (44, 49, 51 and 55 years of age, respectively).

Conclusions:
The most recent guidelines, which lower the PLT threshold to 100 Gi/L from 150 Gi/L for the investigation of causes of thrombocytopenia, reduce the diagnostic rate of MDS. In our retrospective review, 65% of patients would not have had an early diagnosis of MDS. Furthermore, BM aspirate should be considered irrespective of age, since one third of the patients in our case review had MDS with PLT > 100 Gi/L as a single cytopenia and age under 60.

Disclosures:
No relevant conflicts of interest to declare.
Analysis of Related Factors of Malignant Clone Burden of Patients with Myelodysplastic Syndromes
Authors: Shao Zonghong , Yue Lanzhu , Fu Rong , Wang Huaquan , Li Lijuan , Ruan Erbao , Qu Wen , Liang Yong , Wang Guojin , Wang Xiaoming , Liu Hong , Song Jia , Wu Yuhong , Xing Limin , Wang Jun , Guan Jing 
Published: October 2019

Abstract
Abstract 4979

Objective
To investigate the related factors of the malignant clone burden of patients with Myelodysplastic syndromes (MDS), and explore the risk factors of cytogenetic evolution of MDS.

Methods
Seventy-three cases of MDS patients were enrolled in this study who received treatment in the hematological department of General Hospital of Tianjin Medical University from 2004 to 2009. Statistics methods such as t-test, Chi-square test and Logistic regression analysis were used to investigate the correlation between the malignant clone burden and its related factors such as bone marrow dysplasia, peripheral blood and iron metabolism indexes. Furthermore, the risk factors of cytogenetic evolution of MDS were also analyzed.

Results
Odd number-nucleus erythrocytes, double-nucleus granulocytes, hypolobated neutrophils were significantly correlated to high malignant clone burden (P<0.05). PAS positive patients and patients with dysplasia in three myeloid lineages, blasts in peripheral blood exhibited significantly higher malignant clone burden (P<0.05). Patients with abnormal karyotype presented significantly higher level of serum ferritin and lower level of unsaturated iron binding capacity (UIBC) than those with normal karyotype (P<0.05). Odd number-nucleus erythrocytes, megaloblastic granulocytes and high myeloid differentiation index (DI) are risk factors indicating cytogenetic evolution of MDS patients.

Conclusion
The indexes of high malignant clone burden of MDS include: odd number-nucleus erythrocytes, double-nucleus granulocytes, hypolobated neutrophils, dysplasia in three myeloid lineages, blasts in peripheral blood, positive PAS, high level of serum ferritin and low level of UIBC. The risk factors of cytogenetic evolution of MDS include: odd number-nucleus erythrocytes, megaloblastic granulocytes and high myeloid DI.

Disclosures:
No relevant conflicts of interest to declare.
Bone Mineral Density Changes In Patients with Paraproteinemia After Treatment with Bortezomib
Authors: Berno Tamara , Boucher Kenneth , Zhan Fenghuang , Tricot Guido J. , Mughal Benjamin , Zangari Maurizio 
Published: October 2019

Abstract
Abstract 4989

Background:
Bone disease is present at diagnosis in almost all patients with multiple myeloma (MM) and can impact substantially on patient morbidity and quality of life. Decreased bone mineral density is also observed not only in MM but also in patients with monoclonal gammopathy of undetermined significance (MGUS). The pathogenesis of bone disease in MM is complex. The activity of proteasome inhibitor bortezomib has been linked to increased bone formation and osteoblastic activation. Evidence from the available clinical data indicates that bortezomib has a positive impact on bone health in MM and demonstrates a bone anabolic effect.

Methods:
We analyzed retrospectively 53 patients with MM and 16 with MGUS who have completed bone density at least at diagnosis. 21 patients have completed two bone density (3 MGUS and 18 MM). The bone density was obtained in all patients at baseline and in 16 patients repeated after bortezomib treatement with a median time of bortezomib exposure of 6 months. We analyzed T-score values at lumbar spine and at femoral neck.

Results:
With a median age of 66 years, 41 male and 28 female were analyzed. At baseline the mean lumbar spine T-score of all subjects and of 16 MM treated with bortezomib was -0.50 and -0.76 respectively. At baseline the mean femoral neck T-score for all subjects and for 16 MM treated with Bortezomib was -1.56 and -1.31 respectively. The baseline mean lumbar spine T-score for MGUS and MM was -0.71 and -0.43 respectively. The baseline mean femoral neck T-score of MGUS and MM was -1.61 and -1.54 respectively. In the group of 16 patients treated with Bortezomib we observed from baseline a change in lumbar bone mineral density T-score of 0.36 and at femoral neck bone density T-score of 0.25.

Conclusion:
These data show that patients treated with proteasome inhibitor showed moderate increment in bone mineral density at lumbar spine and at femoral neck.

Disclosures:
No relevant conflicts of interest to declare.
IDH1 and IDH2 mutations are Not Frequent In Multiple Myeloma
Authors: Langer Christian , Ibañez Mariam , Liebisch Peter , Zenz Thorsten , Knop Stefan , Einsele Hermann , Paschka Peter , Doehner Hartmut , Doehner Konstanze 
Published: October 2019

Abstract
Abstract 4992
Multiple myeloma (MM) is characterized by frequent and complex genomic abnormalities. However, most genetic abnormalities are already present in the precursor state of MM, the monoclonal gammopathy of undetermined significance (MGUS). Therefore, it is likely that secondary genetic events might contribute to the development from MGUS to symptomatic MM. Mutations in isocitrate dehydrogenase 1 (IDH1) and 2 (IDH2) genes have recently been described as relatively frequent molecular lesions in gliomas and in acute myeloid leukemia (AML). However no larger study has so far examined the frequency of IDH1 and IDH2 gene mutations in patients (pts) with MM.

Methods:
In 188 pts with MM genomic DNA from CD138 sorted plasma cells was used for analyses. Exon 4 of both IDH1 and IDH2 were amplified by PCR and the amplicons were analyzed using a combination of denaturing high-performance liquid chromatography and DNA sequencing. All patients were also characterized by a comprehensive set of FISH probes for the presence of recurring cytogenetic abnormalities.

Results:
185 out of 188 samples were evaluable for analyses. One missense mutation in the IDH2 gene (c.G419A) was identified in the cohort of 185 MM pts (0.5%). This mutation was described as the most frequent IDH2 mutation in AML and is predicted to cause an amino acid change from arginine to glutamin at position 140 (p.R140Q). On cytogenetic analysis this patient harbored a translocation t(11;14) resulting in aberrant expression of CCND1. Additionally, in 15 pts (8%) the recently described single nucleotide polymorphism (SNP) in the IDH1 gene (rs11554137) was detected that has been reported as an adverse prognostic factor in cytogenetically normal AML.

Summary:
Mutations in the IDH1/2 genes are a rare event in MM (0.5%). Further studies are warranted to address the issue if IDH1/2 mutations are restricted to distinct genetic subgroups as for example the group of MM pts with translocation t(11;14).

Disclosures:
No relevant conflicts of interest to declare.
Multi-Parameter Flow Cytometry(FC) Can Detect Proliferating Myeloma Cells After Treatment: Implications for Targeting the Myeloma Proliferative Component
Authors: Lust John , Kumar Shaji , Timm Michael , Donovan Kathleen , Greipp Philip R. , Morice William 
Published: October 2019

Abstract
Abstract 4995

Background:
Multiple myeloma results from an accumulation of monoclonal nonproliferating plasma cells arising from a small subpopulation of proliferating myeloma cells. In an effort to optimize detection of the human myeloma cell growth fraction and obviate the need for slide review, a novel FC strategy was developed combining elements of light chain restriction, surface antigen expression, and ploidy analysis.

Methods:
Bone marrow cells from 41 patients with plasma cell proliferative disorders were lysed with ACK and resuspended in 3% BSA. Cells were stained using a 6-color assay with anti-CD45, anti-CD38, anti-CD138, and anti-CD19. Cells were washed and 100 ul Caltag solution A was added for 15 min. Cells were washed and 100 ul of Caltag solution B, anti-kappa and anti-lambda were added for 10 min. Cells were washed and 100 ul PBS and 3 ul RNAse are added for 15 min. Cells were washed and 400 ul of a 1:1000 solution of 3uM DAPI in Tris 0.1% NP-40 was added. Cells were incubated at 4°C for 45 minutes before running on a FACSCanto instrument for ploidy determination. All patients were analyzed both by flow cytometry and the slide based plasma cell labeling index. Results: Forty-one patients were studied; 34 demonstrated a proliferative fraction and 7 had too few plasma cells for analysis after therapy. Of the 34 patients, 6 had MGUS/SMM, 5 newly diagnosed MM, 7 amyloid, and 16 were treated MM. The mean percent proliferating cells were 1.1% (range 0 – 8.6%) with PCLI and 1.4% (range 0.1 – 12.7%) by flow. The correlation between PCLI and flow gave a RSquare value of 0.54. Twelve patients with a PCLI of 0% had a flow proliferation between 0.1 – 1.4% (mean 0.49%). Treated patients received lenalidomide, dexamethasone, bortezomib, and/or autologous transplantation. All 16 treated myeloma patients with adequate plasma cells had a flow proliferation between 0.2 – 12.7% (mean 2.2%). Conclusion: Flow cytometry offers a useful way to detect the proliferative myeloma component at diagnosis and after treatment. The continued presence of proliferating myeloma cells after treatment may explain why most patients relapse and offers another important marker to monitor and cell population to target in patients with active disease.

Disclosures:
Kumar: Celgene: Consultancy, Research Funding; Millennium: Research Funding; Merck: Consultancy, Research Funding; Novartis: Research Funding; Genzyme: Consultancy, Research Funding; Cephalon: Research Funding.
Multi-Organ Involvement of Multiple Myeloma Cells In the Cases of Recent 10 Years: A Clinicopathologic Study of 81 Consecutively Autopsied Cases
Authors: Hagiwara Shotaro , Mochizuki Makoto , Endo Hisako , Hirai Risen , Tanimura Akira , Takeshita Masataka , Murakami Hirokazu , Miwa Akiyoshi 
Published: October 2019

Abstract
Abstract 5004

Background:
Despite recent progress in treatment, multiple myeloma is still uncurable disease. The impact of modern therapy on the causes of death and pathological feature of end-stage myeloma is not fully understood.

Methods:
We studied autopsied cases with multiple myeloma between 1979 and 2009 at National Medical Center of Global Health and Medicine, Tokyo, Japan. We compared the clinicopathological feature of the autopsied cases in recent 10 years with the cases before 2000. Statistical analysis was performed using student's t-test and chi-square test.

Results:
There were 81 autopsied cases between 1979 and 2009. 31 cases were autopsied in recent 10 years and the older 50 cases were before 2000. Mean age at death was 59.2 and 65.1 years old, and the mean duration of illness was 46.1 and 31.9 months, respectively. Stem cell transplantation was performed in 13 (12 autologous, 1 allogeneic) of recent cases and 3 (2 autologous and 1 allogeneic) of older cases. In recent cases, five patients were treated with bortezomib, 2 were with thalidomide and 2 were with both. Extramedullary infiltration of myeloma cells were observed in both groups. The frequent sites of involvement were spleen, liver, kidney, lymph nodes, lung, pancreas, adrenal gland and perioneum. The infiltration in liver and lung was significantly frequent in the recent cases than in the older cases (58.1% vs. 28.0%, p=0.007, and 38.7% vs. 18%, p=0.039). Infection as a cause of death was noted more frequently in the recent cases than in the older cases (41.9% vs. 18.0%, p=0.019). Amyloid deposition was detected in 16.1% and 22.0% (ns.), and myeloma kidney was noted in 48.4% and 60% (ns.).

Conclusion:
High dose chemotherapy with stem cell support and novel agents have been contributed to improving the survival. However, the increase in resistant bacterial and fungal infection is serious problem. Also, extramedullary relapse after autologous and allogeneic stem cell transplantation is not rare, and extramedullary progression under thalidomide has been reported. In our recent autopsied cases, the incidence of fatal infection and extramedullary involvement of myeloma cells was significantly higher than in the older cases.

Disclosures:
No relevant conflicts of interest to declare.
The Autophagy Gene ULK1 Plays a Role In AML Differentiation and Is Negatively Regulated by the Oncogenic MicroRNA 106a
Authors: Tschan Mario P. , Jost Melanie , Batliner Jasmin , Fey Martin F. 
Published: October 2019

Abstract
Abstract 503
Autophagy is a degradative mechanism mainly involved in the recycling and turnover of cytoplasmic constituents allowing for example survival upon a variety of stress stimuli. Although often referred to as type II programmed cell death, autophagy is primarily a pro-survival mechanism rather than an important cell death effector. During autophagy, part of the cytoplasm or entire organelles are sequestered into double-membraned vesicles, called autophagosomes, that ultimately fuse with lysosomes and degrade their contents. Regarding hematopoiesis there is accumulating evidence that autophagic mechanisms are operative in the development of different hematopoietic lineages. To identify ATG genes with a function in neutrophil differentiation, we employed an shRNA library targeting autophagy regulators in acute myeloid leukemia (AML) cells as a model for neutrophil differentiation. Among others, we found that inhibiting the key autophagy kinase ULK1 (also known as ATG1) using RNAi significantly attenuated all-trans retinoic acid (ATRA)-induced neutrophil differentiation of NB4 and HL60 AML cells. In agreement, we observed significantly elevated ULK1 mRNA and protein levels during neutrophil differentiation of these cells. Moreover, ULK1 expression was significantly reduced in primary AML patient samples as compared to mature myeloid cells from healthy donors. Given the aberrant microRNAs (miRNAs) expression in myeloid malignancies, we wondered if altered miRNA expression in AML might contribute to the low ULK1 levels seen. MiRNAs are 19–25 nucleotide, non-protein coding RNAs that regulate gene expression by directly binding to partially complementary sequences in the 3′-untranslated regions (3′-UTR) of their target mRNAs. Several target prediction software's identified a putative miR-106a binding site in the ULK1 3′-UTR. Interestingly, miR-106a is part of a highly oncogenic miRNA family. We then confirmed direct inhibition of ULK1 by miR-106a using wild-type and miR-106a mutated 3′-UTR ULK1 reporters. Moreover, a decrease of miR-106a expression was seen during ATRA-induced differentiation of AML cells paralleled by increased ULK1 mRNA levels. Lastly, stable overexpression of miR-106a in NB4 cells resulted in decreased ULK1 levels and markedly impaired neutrophil differentiation upon ATRA treatment. In summary, we show for the first time a significant involvement of the autophagy gene ULK1 in neutrophil differentiation and an association of low ULK1 levels with primary AML. Our studies indicate that ULK1 is a putative tumor suppressor in AML that is targeted by the oncogenic miR-106a.

Disclosures:
No relevant conflicts of interest to declare.
A Multicenter Randomized Phase II Trial of Mapatumumab, a TRAIL-R1 Agonist Monoclonal Antibody, In Combination with Bortezomib In Patients with Relapsed/Refractory Multiple Myeloma (MM)
Authors: Belch Andrew , Sharma Atul , Spencer Andrew , Tarantolo Stefano , Bahlis Nizar J , Doval Dinesh , Gallant Gilles , Kumm Elizabeth , Klein Jerry , Chanan-Khan Asher A. 
Published: October 2019

Abstract
Abstract 5031
Mapatumumab, a fully human monoclonal antibody, targets and activates the TRAIL-R1 receptor. We conducted this randomized, controlled phase II trial to evaluate the efficacy and safety of mapatumumab in combination with bortezomib in subjects with relapsed/refractory MM. Response was evaluated using the Bladé criteria. Patients were required to have a measurable serum and/or urine M-protein and have failed 1 or 2 prior therapies for MM. Patients were excluded if they had received prior bortezomib. Patients were randomly assigned to Arm A, 1.3 mg/m2 bortezomib (days 1, 4, 8, 11) every 21 days; Arm B10, bortezomib + 10 mg/kg mapatumumab (day 1) every 21 days; or Arm B20, bortezomib + 20 mg/kg mapatumumab (day 1) every 21 days. Cycles were repeated every 21 days for a maximum of 17 cycles (1 year), progressive disease or unacceptable toxicity. Subjects with complete response (CR) were treated for an additional 2 cycles after documentation of CR (not to exceed 17) and then followed. A total of 104 subjects from 4 countries were randomly assigned to the treatment arms. The addition of mapatumumab to bortezomib did not increase the response rate, progression-free survival (PFS) or duration of response compared to bortezomib alone. Adverse events were generally balanced across treatment groups; there was no evidence that mapatumumab exacerbated toxicities associated with bortezomib. As expected, the most common toxicities were hematological and peripheral neuropathies. The results do not support further evaluation of mapatumumab in combination with bortezomib in patients with advanced MM. Additional trials of mapatumumab in other indications are on-going.
Arm A (n= 35) Arm B10 (n=33) Arm B20 (n=36) CR 0 0 2 PR 18 10 17 CR + PR n (%) 18 (51.4) 10 (30.3) p=0.08a 19 (52.8) p=0.91b Median PFS mo (95% CI) 8.7 (7.6, 10.0) 4.7 (2.5, 7.4) p=0.29a 5.7 (5.2, 8.9) p=0.21b Median Duration of Response mo (range) 8.5 (6.9, 14.2) 9.3 (3.9, 16.1) p=0.92a 7.6 (4.3, 8.8) p=0.41b a P value for comparison of Arm A with Arm B10 b P value for comparison of Arm A with Arm B20

Disclosures:
Gallant: Human Genome Sciences, Inc.: Employment, Equity Ownership. Kumm:Human Genome Sciences, Inc.: Employment, Equity Ownership. Klein:Human Genome Sciences, Inc.: Employment, Equity Ownership.
T Regulator Cells (Treg) In Patients with Myelofibrosis
Authors: Shakil Shams A , Chen Chi , Surahio Ali , Mirza Mariam B , Su Marisa Y , Sun Yiwu , Wang Jen-Chin 
Published: October 2019

Abstract
Abstract 5051
Primary myelofibrosis (PMF) and myelofibrosis post essential thrombocytosis (MF-ET) or myelofibrosis post polycythemia vera (MF-PV) have been reported to be associated with autoimmune phenomenon, such as Coomb positive anemia, lupus anticoagulant, positive ANA and the presence of circulating immune complex etc. Regulatory T cells (Treg) also have known to play important roles in modulating immune responses. Therefore we studied Treg cells in 25 patients with MF including PMF (12), MF-ET (8), MF-PV (5) and compared with other MPD including ET (7), PV (8), and normal volunteer controls (17). Mononuclear cells (MNC) were separated from peripheral blood. 106 MNCs were stained for flow cytometry analysis using Treg Detection Kit from Miltenyi Biotec Inc (Auburn, CA). The numbers of Treg was calculated as the percentage of positive CD4+ CD25+ FoxP3+ T cells (Treg) from the numbers of gated CD4+ cells. Treg function was evaluated by XTT cell proliferation assay (Invitrogen) with ratios of Treg vs. T-effector cells (CD4+ CD25-) at 1:1, 1:2, 1:4, and 0:1 in the presence of anti-CD3 CD28 microbeads (Invitrogen). The results (mean ± SE) showed numbers of Treg cells in MF were 0.73 + 0.08, other MPD were 1.37 ± 0.22, and normal controls were 0.96 ± 0.27 (p=NS). Treg function was evaluated in 12 patients with MF and 6 normal volunteer controls. Four MF patients were found to have significant lower values than controls. We concluded that in MF, quantitatively Treg were not different from other MPD or normal controls but Treg dysfunction was observed in 30–40 % of MF patients. This could explain why some patients with MF are prone to develop autoimmune phenomenon. Further studies with more patients are in progress.

Disclosures:
No relevant conflicts of interest to declare.