bluuebunny/crossref_metadata_2025 huggingface dataset:
Myelodysplasia (MDS) patients' Experiences with Blood Transfusions
Authors: Twiss James , McKenna Stephen P , Crawford Sigrid Rebecca , Wilburn Jeanette N , Loth Kevin , Mufti Ghulam J
Published: October 2019

Abstract 5033

Introduction:

Although the main treatment for MDS is blood transfusions, scarce information is available on patients' experiences of the treatment. Only one relevant study could be identified which reported that patients have a preference for being transfusion independent (Szende et al. Valuation of transfusion-free living in MDS: results of health utility interviews with patients. (2009). Health Qual Life Outcomes, 7:81). As part of a study investigating the effect of MDS and its treatment on quality of life, patients were asked about their experiences with transfusions.

Methods:

Qualitative, unstructured interviews were conducted with MDS patients. Verbatim transcripts were produced and subjected to Interpretative Phenomenological Analysis (IPA). IPA aims at gaining an understanding of patients' experiences from their own perspective. Themes are generated based on issues identified during the interview process.

Results:

The sample included 23 patients with blood transfusion experience (male = 11 (47.8%); mean (SD) Age=65.3 (11.2)). Of the patients interviewed, 9 (39.1%) were currently receiving regular transfusions, 5 (21.7%) had been transfusion dependent previously and 9 (39.1%) had some transfusion experience. Illness information for these interviewees is shown in the Table 1. Five major themes were identified in the analyses; positive aspects of transfusions, the cyclic nature of symptoms, transfusion-related limitations on functioning, associated iron chelation therapy and problems with actually receiving the transfusions. Transfusions were perceived positively by some patients as they relieve patients' everyday symptoms. However, these benefits diminish between transfusions leading to cycles of good and poor symptoms, functioning and psychological distress. Limitations to patients' lives resulted from frequent and often long trips to hospital. Patients reported negative experiences with the iron chelating drug Deferrioxamine which is needed to reduce iron poisoning. The drug is generally administered subcutaneously by means of a battery operated pump and is usually worn for several hours a day causing pain and discomfort. The transfusions were also reported to be boring, uncomfortable and at times painful.

Conclusions:

Transfusions can have a large negative impact on patient's lives. The results suggest there is a need for alternative forms of treatment for patients with MDS. Further quantitative work assessing patients' experiences with transfusions is required.

Disclosures:

Twiss: Celege Ltd: Consultancy, Research Funding. McKenna:Celgene Ltd: Consultancy, Research Funding. Crawford:Celegene: Consultancy, Research Funding. Wilburn:Celegene Ltd: Consultancy, Research Funding. Loth:Celgene Ltd: Employment.
Expression of dlk1 in the Bone Marrow Cells of Patients with Myelodysplastic Syndrome and Its Clinical Significance
Authors: Shao Zonghong , Yue Lanzhu , Fu Rong , Li Lijuan , Ruan Erbao , Qu Wen , Liang Yong , Wang Guojin , Wang Xiaoming , Liu Hong , Song Jia , Wu Yuhong , Xing Limin , Wang Jun , Guan Jing , Wang Huaquan
Published: October 2019

Abstract 5042

Objective

To investigate the expression of dlk1 gene (delta-like 1) in the bone marrow cells of patients with Myelodysplastic syndrome (MDS), and explore the molecular marker for early diagnosis of MDS.

Methods

The expression of dlk1 mRNA in the bone marrow cells of cases with MDS, AML and normal controls were measured by RT-PCR, aiming to search for the cytogenetic marker of MDS malignant clone.

Results

The expression of dlk1 mRNA in bone marrow cells of MDS patients (0.7342±0.3652) was significantly higher than that of normal controls (0.4801±0.1759) (P<0.05), and was significantly positively correlated with the proportion of bone marrow blasts(r=0.467,P<0.05). The expression of dlk1 mRNA significantly increased as the subtype of MDS advanced (P<0.05). Patients with abnormal karyotypes displayed significantly higher expression of dlk1 mRNA (0.9007±0.4334) than those with normal karyotypes (0.6411±0.2630) (P<0.05). Patients with higher expression of dlk1(≥0.8) presented significantly higher malignant clone burden (0.4134±0.3999) than those with lower expression (<0.8) of dlk1 (0.1517±0.3109) (P<0.05).

Conclusion

dlk1 gene was highly expressed in MDS patients, which increased as the subtype of MDS advanced. The expression of dlk1 mRNA was significantly positively correlated with the proportion of bone marrow blasts. High expression of dlk1 gene suggests high malignant clone burden of MDS.

Disclosures:

No relevant conflicts of interest to declare.
Methylation Analysis of the Wnt Antagonist in Myelodysplastic Syndrome
Authors: Tong Hongyan , Mei Chen , Li Kongfei , Jin Jie
Published: October 2019

Abstract 5048

Background and Objectives:

Myelodysplastic syndrome (MDS) is one of the most threatening hematological malignancies. Recently, the epigenetic changes have been recognized in the MDS, and some research found that the aberrant DNA methylation had close relationship with the MDS. Meanwhile, some studies showed that the activation of the Wnt signaling pathway and abnormal methylation of the Wnt antagonists had close relationship with hematological malignancies, such as AML AALL Aand CLL, but less is known in MDS. In our research, we studied the methylation status of Wnt antagonists (DKK1, DKK3, HDPR1, WIF-1, SFRP1 and SFRP4) in the patients with MDS and evaluated the role of them in the pathogenesis and progression of MDS, with providing a new theory support for clarifying the complicated pathogenesis and progression of MDS.

Methods:

The methylation status of Wnt antagonists (DKK1, DKK3, HDPR1, WIF-1, SFRP1 and SFRP4) in pretreatment bone marrow samples from 53 patients with MDS was measured by methylation-specific polymerase chain reaction (MSP). On the other hand, we collected the clinical materials of the patients with MDS and follow up the patients. Then the correlation between methylation and clinical features as well as prognosis of MDS patients was analyzed byχ2 test and Kaplan-Meier method. Results: In 53 bone marrow samples, the methylation frequencies of the Wnt antagonists were as follows: SFRP4 for 62.3 % (33/53), DKK1 for 45.3 % (24/53), HDPR1 for 34.0 % (18/53), SFRP1 for 15.1 % (8/53), DKK3 for 9.4 % (5/53), and WIF-1 for 5.7 % (3/53). After analyzing individual tumor suppressor gene, clinical parameters and prognostic information, it was found that the patients with the percentage of BM blast above 5% had higher methylation frequency of HDPR1 than the patients with the percentage of BM blast bellow 5% (44.4%∼a13.3%, P=0.034); besides, the methylation frequency of HDPR1 exhibited significant differences in the MDS subtypes (P=0.019), and was significantly correlated with the WPSS (P=0.037); In addition, Kaplan-Meier survival curve indicated that the mean overall survival of patients with methylation was significantly shorter than that of patients without HDPR1 methylation (457.3 days vs. 919.6 days, P=0.037) (Fig.1).

Conclusion:

The methylation of the Wnt antagonists (DKK1, DKK3, HDPR1, WIF-1, SFRP1 and SFRP4) were observed in patients with MDS and their methylation frequencies were different. The aberrant methylation of HDPR1 may play an important role in the progression and prognosis of MDS.

Disclosures:

No relevant conflicts of interest to declare.
Multiple Myeloma with Secretory and Non-Secretory Biclonal Gammopathy
Authors: Vinayek Namita , Kloecker Goetz H , Riley Beth C.
Published: October 2019

Abstract 5092

Multiple myeloma (MM) accounts for 10 % of hematological malignancies. MM with biclonal gammopathy are rare and seen in 1 % of all MM cases. In 99% of the MM cases, paraproteins are secreted in the serum and/or urine, in the remaining 1% the paraproteins are synthesized but not secreted. Only 2 % of patients are younger than 40 years of age.

We report a 36 year old African American female who presented with diffuse lytic lesions. Serum electrophoresis (SPEP) revealed IgG kappa M protein of 3.8 g/dl and a IgG Kappa M spike of 2.6 mg/dl. Free light chains (SFLC) revealed elevated free kappa chains of 500 mg/dl, elevated free lambda chains of 928 mg/dl and K/L ratio of 0.55. Urine protein UPEP showed an M spike of IgG kappa and free kappa light chains. The B2 microglobulin was 26.8 mg/dl. Patient's recent T10 Bone biopsy done at outside hospital had a near complete replacement of the marrow by plasma cells which were biclonal plasma cells by IHC, one kappa restricted and one lambda restricted. Since the IFE showed only a single IgG kappa M protein spike, the lambda plasma clone was apparently non-secretory.

Treatment was started with cyclophosphamide, bortezomib and dexamethasone. After 22 days of treatment, the M protein has decreased 1.39 mg/dl. A repeat SFLC also shows further decline in free kappa light chains. As the IFE showed one gamma M spike and bone biopsy had two clonal plasma cells it was concluded that one of the neoplasms is nonsecretory. Although the elevated lambda free light chain in the serum is now does point towards a second plasma cell neoplasm.

Biclonal gammopathy is rare and accounts for 1% of all MM cases. To our knowledge, this is the first reported case of a biclonal, secretory and nonsecretory, gammopathy

Disclosures:

No relevant conflicts of interest to declare.
The Nestin Expression in Monoclonal Gammopathies
Authors: Kovarova Lucie , Svachova Hana , Muthu Raja Karthick Raja , Varmuzova Tamara , Zarbochova Pavla , Penka Miroslav , Hajek Roman
Published: October 2019

Abstract 5111

Background.

Monoclonal gammopathies (MG) are the most common plasma cell disorders characteristic by a presence of clonal plasma cells (PCs). The nestin is considered to be a marker of multipotent proliferative precursors, but recently its expression was found in PCs of multiple myeloma (MM) as well. Consequences of the nestin expression still remain unclear.

Aim.

Phenotypic analysis of the nestin expression in the most frequent MGs and its correlation with other markers.

Patients and Methods.

Total of 45 samples was analyzed [31 newly diagnosed untreated MM patients, 14 untreated monoclonal gammopathy of undetermined significance (MGUS) subjects and 5 non-MG samples]. Whole bone marrow CD38+CD138+ PCs were studied for intracellular expression of nestin and surface expression of CD19, CD20, CD27, CD28, CD33, CD56, and CD117 by flow cytometry.

Results.

Flow cytometric analysis shown the nestin expression in 21.4 % (3/14) of MGUS and 45.2 % (14/31) of MM patients, on the other hand no expression of nestin was found in non-MG controls. Expression of the nestin was relatively homogeneous, but not highly intensive. The nestin positive PCs did not express CD19 and/or CD20, but expression of CD27 was found in 29.4 % (5/17) cases, CD28 in 11.8 % (2/17), CD33 in 5.9 % (1/17), CD44 in 35.3 % (6/17), CD56 in 88.2 % (15/17), and CD117 in 29.4 % (5/17) cases. Statistical analysis did not found any significant correlation and/or difference in phenotype when compared the nestin positive and negative samples, although the nestin negative samples expressed CD56 less frequently then the nestin positive samples [46.4 % (13/28) vs. 88.2 % (15/17)], respectively.

Summary/Conclusion.

Our result showed difference in the nestin expression when compared bone marrow of MGUS subjects with MM patients. Although there was no correlation between the nestin positivity and phenotype of PCs, we suppose that the nestin could associate with unfavorable prognosis but further studies are needed to elucidate its clinical implication in MGs.

Disclosures:

No relevant conflicts of interest to declare.
A Novel Small Molecule with Epigenetic Activity Prolongs Survival in a Mouse Model of Multiple Myeloma Refractory to Standard Drugs
Authors: Vatolin Sergei , Shahper Khan Nazeer , Parker Yvonne , Lindner Daniel , Reu Frederic J.
Published: October 2019

Abstract 5121

Multiple myeloma refractory to bortezomib, IMiDs™, and conventional therapies represents an unmet medical need. An increasing number of patients progress to this stage since treatment related mortality has decreased. To test promising compounds for activity in this setting we established an NSG mouse xenograft model with serial transplantation by tail vein injection of myeloma cells from a patient with IgG kappa myeloma relapsed and refractory to all standard drugs. Eight days after tail vein injection monoclonal human IgG can be detected in serum. Bone marrow engraftment in young (6–12 weeks) NSG mice after sublethal radiation (275cGy) is close to 100% (n=32). Untreated mice die within less than 2 months, usually with liver and spleen metastasis (anti-human CD138 flow cytometry). In a drug screen that used a novel method developed in our lab, chromatin condensation PCR, we identified a non nucleoside compound (4I3) that potently (1mM) reactivated expression of epigenetically silenced genes and displayed cancer-specific growth and survival inhibition in myeloma cell lines but not normal cells. Normal bone marrow cells continued to divide at doses 10x higher than required to kill 80% of myeloma cells. 4I3 suppressed DNMT1 protein but rapid cell kill (within 1–2 days) suggested additional mechanisms which we currently investigate. Given IV to mice after documentation of engraftment by IgG serum immunoblots, it prolonged survival in an ongoing experiment. Updated results will be presented at the meeting.

Disclosures:

No relevant conflicts of interest to declare.
JAK2 Mutation Analysis and Function of Bone Marrow Mesenchymal Stem Cells in Myeloproliferative Neoplasms
Authors: Tian Hong , Xu Yang , Chen Guanghua , Qiao Man , Depei Wu
Published: October 2019

Abstract 5179

Background:

JAK2V617F and JAK2 exon12 mutations in haematopoietic cells were partially responsible for the pathogenesis of myeloproliferative neoplasms (MPN).But it was still unclear whether bone marrow mesenchymal stem cells (BMSCs), the significant component of hemopoiesis microenvironment, were participated in the pathogenesis of MPN.

Objective:

To study the physiopathology characteristics and analyze JAK2 mutation in BMSCs from MPN patients.

Methods:

By searched for the JAK2V617F mutation and exon 12 mutation in 135 MPN patients' blood /bone marrow samples, 20 patients with JAK2V617F mutation, 10 patients with JAK2 exon 12 mutation, 5 JAK2-mutation-negetive patients and 10 healthy donors were recruited. The phenotype, mesenchymal differentiation capacity, expression of hematopoietic and immune molecules and JAK2 mutation of isolated bone marrow BMSCs were detected.

Results:

BMSCs derived from the four groups were found to be similar in morphology, differentiation ability and expression of hematopoietic and immune molecules. Primary study indicated that the isolated BMSCs from patients groups were not able to harbor JAK2 mutation in spite of positive or negative JAK2 mutation in blood /bone marrow samples.

Conclusion:

BMSCs from MPN patients had similar biological characteristics to healthy donors, and BMSCs were not likely involved in pathogenesis of MPN.

Disclosures:

No relevant conflicts of interest to declare.
Selective Disruption of PU.1 in Mature Dendritic Cells Affects Their Tissue Distribution and T Cell Homeostasis
Authors: Iino Tadafumi , Iwasaki Hiromi , Kohno Kentaro , Mizuno Shin-ichi , Arinobu Yojiro , Tenen Daniel G. , Reizis Boris , Akashi Koichi
Published: October 2019

Abstract 518

PU.1, a hematopoietic transcription factor, is indispensable for development of conventional dendritic cells (cDCs) from hematopoietic stem cells. However, the function of PU.1 in mature cDC remains unclear. To test the possible role of PU.1 in mature cDCs, we developed mice lacking PU.1 selectively in mature cDCs (DC-PU.1D/D mice) by crossing a PU.1flox mouse line with a transgenic Itgax (CD11c)-Cre strain. In these mice, cDCs were dramatically reduced in spleen, thymus, lymph node, and skin, down to <40%, <25%, <10% and <5% of DCs in control mice respectively, whereas bone marrow cDCs and common dendritic cells progenitors (CDPs) were not affected. Surprisingly, T cell numbers were significantly decreased in DC-PU.1D/D mice, whereas thymic T cell development was normal, suggesting that maintenance of mature T cell pool might be impaired, presumably by dysfunction of PU.1D/D cDCs. In fact, PU.1D/D cDCs failed to efficiently induce ovalbumin-specific T cell response and to produce inflammatory cytokines in response to Toll like receptor (TLR) stimulation both in vitro and in vivo. The intravenous transfer of spleen PU.1D/D cDCs failed to repopulate the spleen of recipient mice, suggesting their poor survival in vivo. Furthermore, the expression of critical molecules for inflammatory responses was downregulated in PU.1D/D cDCs as compared to normal cDCs. These molecules included Myd88 and NFkB that are downstream molecules of TLR signaling, CD86 that is required for T cell stimulation, and CCR7 that is required for cDC migration. These results clearly show that PU.1 is required for development of the functional cDC pool, and the cDC pool plays a critical role in T cell homeostasis.

Disclosures:

No relevant conflicts of interest to declare.
Characterizing Canine Lymphoma As a Potential Large Animal Model of Human Diffuse Large B-Cell Lymphoma
Authors: Richards Kristy L. , Motsinger-Reif Alison , Chen Hsiao-Wei , Fedoriw Yuri D. , Fan Cheng , Nielsen Dahlia M. , Thomas Rachael , Smith Chris , Dave Sandeep S. , Perou Charles M. , Breen Matthew , Suter Steven E.
Published: October 2019

Abstract 5193

Diffuse large B-cell lymphoma (DLBCL) affects ∼25,000 people in the U.S. each year, and fewer than half of them are cured with standard therapy. DLBCL can be divided into two subtypes by gene expression profiling, germinal center B-cell (GCB) type and activated B-cell (ABC) type. ABC-type DLBCL patients have significantly poorer outcomes. To improve therapeutic options, better animal models that accurately mimic human DLBCL (hDLBCL) are needed. Canine DLBCL is one of the most common cancers in veterinary oncology. Similar to human DLBCL patients, dogs with lymphoma are treated with both CHOP-like regimens and autologous stem cell transplants. Morphologically, canine lymphomas are similar to hDLBCL, with shared histologic markers, such as CD20 and PAX5. With recent technologies based on knowledge of the canine genome sequence, it is now possible to evaluate dogs as a potential large-animal model for hDLBCL.

We evaluated 58 canine B-cell lymphomas by generating comprehensive gene expression profiles and comparing them to previously published hDLBCL expression profiles. Canine B-cell lymphoma expression profiles were similar in some ways to hDLBCLs. For instance, increased expression of NF-kB pathway genes was noted in a subset of lymphomas, mirroring NF-kB pathway activation in human ABC-type DLBCL. Furthermore, immunoglobulin heavy chain (IGH) mutation status, which is correlated with ABC/GCB cell of origin in hDLBCL, separated canine DLBCL into two groups with statistically different progression-free and overall survival times. However, canine DLBCL differed from hDLBCL in other aspects, including rare immunohistochemical positivity for BCL6 and MUM1/IRF4. Collectively, these results define aspects of canine B-cell lymphomas that resemble hDLBCL, identifying molecular similarities that could allow dogs to be used as a representative model of hDLBCL. Further comparative studies, including therapeutic trials, could potentially improve outcomes in both species.

Disclosures:

No relevant conflicts of interest to declare.
Stromal Interaction of Lymphoma Cells Regulates Survival and Chemoresistance
Authors: Park Gyeongsin , Song Byunghoo , Lee Yuji , Lee Ahwon , Chung Yang-Guk , Kang Chang Suk , Oh Il-Hoan
Published: October 2019

Abstract 5195

High-grade lymphomas are aggressive but largely curable, whereas low-grade lymphomas are indolent, but frequently recur to be incurable, paradoxically. Mesenchymal stromal cells (MSCs) have been known to participate for reconstituting microenvironment. Studies show that signalings between normal lymphoid cells and stromal cells were frequently altered in high grade lymphomas, but relatively conserved in low grade lymphomas. However, which cell and mechanism is responsible for lymphoma recurrence remains unclear. Here we hypothesized that the interaction with stroma may play a role for lymphoma cell growth and survival. For this, we investigated the effect of MSCs on lymphoma cell growth and chemo-resistance by using a coculture system with MSCs (derived from tonsil), as a stromal microenvironment for lymphoma cells. First, coculture of lymphoma cell line (Pfeiffer) and primary lymphoma cells with/without MSCs for 3 days showed that the MSC-cocultured cells grew more rapidly (1.4 times and 1.8 times for Pfeiffer and primary cells, respectively) than MSC-free lymphoma cells. To further investigate the underlying mechanism of promoting growth, lymphoma cells were cocultured with MSCs in the presence or absence of transwell filter. At day 4, the proliferation of lymphoma cells in the absence of transwell was 3.5 times higher than in the presence of transwell. Interestingly, the lymphoma cells cocultured with MSCs showed increased expression of CXCR4 compared with lymphoma cells cultured alone. When the cyto-protective effect of MSCs was examined by doxorubicin treatment (1ug/ml) in the presence or absence of MSCs, 91% of MSC-cocultured cells survived, whereas only 28% of stroma-free cultured cells survived. These results demonstrate that the direct contact interaction with stroma might be important for lymphoma cell growth and survival.

In conclusion, our data suggest that the interaction with stromal microenvironment is an important factor for survival of lymphoma cells which cells might be responsible for chemoresistance, and raise the possibility that the stromal interaction could be a potential target for lymphoma treatment.

Disclosures:

No relevant conflicts of interest to declare.

This research was supported by a grant (10172KFDA993) from Korea Food & Drug Administration in 2011.

Disclosures:

No relevant conflicts of interest to declare.
Clinical Significance and Prognosis of Mir-155 and Mir-146a Expression Levels in Formalin-Fixed/Paraffin-Embedded Tissue of Patients with Diffuse Large B-Cell Lymphoma
Authors: Zhong Hua
Published: October 2019

Abstract 5197

Introduction

Diffuse large B cell lymphoma (DLBCL) is an aggressive malignancy that accounts for nearly 40% of all lymphoid tumors. Both miR-155 and miR-146a expression levels are suggested to be regulated by NF-KB expression in DLBCL. In present study, the expression of miR-155 and miR-146a were investigated to evaluate the prognosis in DLBCL patients.

Patients and Methods

The expression levels of miR-155 and miR-146a in formalin-fixed/paraffin-embedded tissue of patients with DLBCL (n=50) have been compared with reactive hyperplasia lymphoid nodes (n=20) by using real-time polymerase chain reaction. Clinical characters of patients such as LDH, β2-MG, International Prognostic Index (IPI), ECOG score and c-myc expression levels have also been investigated.

Results

1) The expression levels of miR-155 and miR-146a were significantly higher in DLBCL patients than control (P<0.001). (Fig 1)

2) The expression levels of miR-155 and miR-146a were different between germinal center B-cell like DLBCL (GCB-DLBCL) and activated B-cell-like DLBCL (ABC-DLBCL), but the difference had no significance (P>0.05).

3) There was correlation between the expression of miR-155 and miR-146a in DLBCL patients (r =0.639, P<0.001).

4) There was positive correlation in the expression of miR-146a with LDH, β2-MG, IPI, ECOG score and c-myc expression levels in DLBCL patients.

5) Lower expression levels of miR-155 and miR-146a were found to be associated with high complete remission (CR) rate and overall response (OR) rate in DLBCL patients (P<0.05).

6) Lower expression levels of miR-146a in DLBCL patients were only associated with better 5 years progression free survival (5y-PFS) (P=0.044), but lower expression levels of miR-155 in DLBCL patients were associated with better 5y-PFS (P=0.013) and 5 years overall survival (5y-OS) (P=0.007).

7) Multivariate analysis using a Cox proportional hazard model confirmed that either miR-155 or miR-146a was the independent predictor. (For miR-155 5y-PFS, Hazards ratio 0.260, 95% CI, 0.058–0.801; for miR-146a 5y-PFS, Hazards ratio 0.251, 95% CI, 0.068–0.922)

8) Compared to CHOP protocol, the higher miR-155 expression level patients who chose R-CHOP protocol for treatment achieved better 3y-PFS and 3y-OS (P<0.05).

Conclusions

The expression levels of miR-155 and miR-146a were higher in DLBCL patients. Moreover, high expressions of these two microRNAs were associated with the disease progression. Patients with higher miR-155 expression levels may be benefit from the protocol including Rituximab.

Disclosures:

No relevant conflicts of interest to declare.
EBV-Negative Primary Pulmonary Lymphoma in a Patient with AIDS
Authors: Storey Raul E. , Mandadi Mounika , Abdolmohammadi Alireza , Natarajan Nagendra , Kloecker Goetz H , Stephen Slone P
Published: October 2019

Abstract 5203

AIDS-related primary pulmonary lymphoma (PPL) is defined as exclusive involvement of the lung by lymphoma with no evidence of extrapulmonary disease at the time of diagnosis and three months thereafter. It is an aggressive disease with a median survival time of four months. A 46 year old male patient with a seven year history of HIV/AIDS and CD4 count of 200/ul was hospitalized because of flank pain, hematuria and hemoptysis. Abdominal CT scan incidentally revealed multiple nodules in the lower lobes of both lungs. Serial chest CT scans demonstrated multiple nodular opacities in all lobes of the lungs that enlarged rapidly and coalesced together. Clinicians favored opportunistic infection due to rapid disease progression but included Wegener's granulomatosis in the differential diagnosis because of hemoptysis. Open lung biopsy showed diffuse large B cell lymphoma (DLBCL), NOS, with an activated B cell phenotype (CD10−/BCL6−/MUM1+) and high proliferative index (Ki67 90%). CD79A expression confirmed B cell lineage. The diagnostic criteria for PPL were fulfilled because staging failed to detect extrapulmonary disease. Infarction due to the angiodestructive nature of the PPL and short doubling time accounted for hemoptysis and rapid disease progression, respectively. The patient died 45 days after admission because of shock, presumably due to superimposed infection. Unlike prior reports of AIDS-related PPL, our case was negative for CD19, CD20 and Epstein-Barr virus (EBV). Although PPL is a rare disease, it deserves consideration in the differential diagnosis of rapidly progressive lung nodules in AIDS patients. To the best of our knowledge, this is the first reported case of EBV-negative PPL.

Disclosures:

No relevant conflicts of interest to declare.
TP53 Mutation Is Rare In Primary Myelofibrosis
Authors: Greaves Wesley O. , Verma Shalini , Bisrat Tigist , Rahimi Hamed , Paladugu Abhaya , Bueso-Ramos Carlos E. , Patel Keyur P. , Luthra Rajyalakshmi , Yao Hui , Verstovsek Srdan
Published: October 2019

Abstract 5241

Introduction

TP53 is the most frequently mutated tumor suppressor gene in human cancers and is usually associated with an aggressive disease course. TP53 mutation has been described in a variety of hematopoietic neoplasms, and has been suggested to play a role in leukemic transformation of myeloproliferative neoplasms (MPN). However, the incidence as well as the clinical and pathogenetic implications of TP53 mutation in each sub-category of MPN, including primary myelofibrosis, have not been described. In this study, we investigated the presence and potential clinical significance of TP53 mutations in a large series of primary myelofibrosis cases.

Patients and Methods

We retrieved archival bone marrow DNA from 51 consecutive patients diagnosed with primary myelofibrosis at The University of Texas MD Anderson Cancer Center between the years 2005 and 2007. Diagnosis was based on morphologic, immunophenotypic, cytogenetic and molecular evaluation of bone marrow in conjunction with clinical data. Only 2 patients had blast counts >10% (11 and 14%). Twenty nine of 50 (58%) patients showed JAK2 p.V617F mutation and all patients were negative for BCR-ABL1 translocation on routine clinical testing. DNA samples were assessed for sequence variation in exons 4 through 9 of TP53 by both high resolution melting curve (HRM) analysis using LightCycler® 480 System (Roche, Indiana IN) and bidirectional Sanger sequencing using 3730XL DNA Analyzer (Life technologies, Carlsbad CA).

Results

The mean overall survival was 5.7 years. Five patients developed acute leukemia, all of whom died of disease. By Sanger sequencing, only one (1.9%) case showed an amino acid-altering mutation in TP53: c.707A>G (TAC to TGC) in codon 236 (p.Y236C) of exon 7. In addition, 8 cases showed silent mutations/single nucleotide polymorphisms of unknown significance - c.36G>A (CCG to CCA) in exon 4 (n=3) and c.213A>G (CGA to CGG) in exon 6 (n=6). The p.R72P polymorphism in exon 4 which has been described in other hematopoietic neoplasms was present in 1 patient. All cases with a mutant sequence by Sanger sequencing also showed a variant melting curve pattern by HRM analysis. The patient with TP53 mutation died 2 years after presentation from progressive myelofibrosis without developing acute leukemia.

Conclusion

TP53 mutation is very rare in primary myelofibrosis.

Disclosures:

No relevant conflicts of interest to declare.
Over-Expression of PAI-1 and TAFI Predict Stroke: Two Faces of Ame Coin
Authors: Corinaldesi Giorgio
Published: October 2019

Abstract 5249

TAFI (thrombin-activated fibrinolysis inhibitor) and PAI-1 play important roles in fibrinolysis, as they both reduce plasmin generation; PAI-1 a 50 KDa glycoprotein is a serine-proteinase inhibitor; TAFI is a 55 KDa plasma zymogen that is activated by either plasmin/thrombin or thrombin/thrombomodulin complex into an active carboxypeptidase, in particular, it is a basic zinc dependent metallo-carboxipeptidase specific for C-terminal lysine and arginine residue inhibiting degradation of the fibrin clot.

We investigated the influence of PAI-1 and TAFI levels on stroke by determining their plasmatic antigen concentration by an electro-immunoassay. We found that PAI-1 levels are increased during the early stage of stroke and that it may also predict clot lysis resistance in patients treated with trombolysis (38.8 ng/ml +/− 14.6, p <0.0001) compared with control subjects (16.2 ng/ml +/−6.8, p <0.0001); TAFI levels were significantly higher in patient with stroke (184.8 nmol/l +/− 62.6 nmol/l), than in healthy control subjects (102.4 nmol/l +/− 36.4 nmol/l, p <0.0001). We have observed that a combination of both PAI-1 higher than 24.6 mg/ml (OR: 12.8. CI: 1.16 to 134.2), and TAFI higher than >180 nmol/l (OR.12.9 CI 1.42 to 116.6), had a sensibility of 92.2%, and a specificity of 98.4% in predicting stroke. The analysis of these results showed that TAFI antigen levels higher than 180nmo/L combined with PAI-1 levels higher than 24 mg/ml are a strong risk factor for clot lysis, stroke (for the evolution, severity, and outcome) and showed a potentially attractive target for fibrinolytic therapies.

Disclosures:

No relevant conflicts of interest to declare.
Metabolic and Inflamatory Proteins Differently Expressed in Platelets From Deep Venous Thrombosis Patients
Authors: Flores Nascimento Mariane Cristina , Kleinfelder-Fontanesi Karina , de Andrade Orsi Fernanda Loureiro , Seeholzer Steven H , Ischiropoulos Harry , Annichino-Bizzacchi Joyce M
Published: October 2019

Abstract 5256

BACKGROUND:

Deep venous thrombosis (DVT) is multi-causal disease associated to a high morbi-mortality due to complications as pulmonary embolism and post-flebitic syndrome. The incidence is about 20 to 30%, and 25% of the patients will present recurrence in 5 years. The identification of new risk factors is important in clinical practice to prevent new thrombotic events. The role of the platelets on DVT is still not well defined.

AIM:

The objective of this study was to analyze the hole proteins profile of platelets obtained from DVT patients and compare to the same matherial derived from healthy controls.

PATIENTS:

peripheral blood samples were collected from 3 spontaneous DVT patients and from 1 sibling and 1 neighbor for each patient in order to minimize the genetic and environmental interferences. These patients presented spontaneous and recurrent episodes of lower limbs proximal DVT and all of them mentioned a familiar history of coagulation disorders.

METHODS:

the platelets were washed, lysed, and the proteins were alkylated, reduced, precipitated with acetone and hydrolyzed by trypsin. 100mg of peptides were then separated by hydrophobicity using HPLC, and 8 fractions were obtained and directed to the LTQ-Orbitrap mass spectrometer. The proteins search was performed by Sorcerer-SEQUEST.

RESULTS:

We identified 5 proteins that were present on patients and absent in all the controls: Apolipoprotein A1 Binding-Protein, Coatomer (z1 sub-unit), Estradiol 11–17-b Dehydrogenase, Leucotriene A-4 Hydrolase and Sorbitol Dehydrogenase. Western-Blotting was performed with specific antibodies and validated the results.

CONCLUSIONS:

with this study it was possible to identify proteins up to date non-related to the physiopathology of DVT, that could be involved with metabolic and inflammatory processes.

Disclosures:

No relevant conflicts of interest to declare.
Hemolytic Intravascular Anemia (HIA) Microangiopathic-Like and Deep Venous Thrombosis (DVT) Due to Ozone Exposure: Case Report
Authors: Cortázar-Benítez Luis Fernando , Viveros Pablo Vargas , Alvarez Alfredo Aiza , Monroy Rafael Hurtado
Published: October 2019

Abstract 5278

HIA due to ozone exposure on patients with glucose 6 phosphate dehydrogenase deficiency (G6PDd) is extremely rare. The purpose of this report is to describe in detail a case of HIA and DVT on a woman without G6PDd, with a successful treatment with fresh frozen plasma (FFP) transfusion and Thrombolysis.

The patient is a 36 years old woman, that 3 months before she was diagnosed with multiple sclerosis (MS) because paresthesias in the fingers of her left foot and she received treatment with blood ozone exposure (at unknown dose) in 3 sessions each week for 3 months. The patient attended to our center with severe anemic syndrome during the last 2 weeks and disabling pain of her left leg of 12 hours of evolution. Physical examination showed pale ++++, jaundice ++, functional systolic murmur grade IV, without adenomegaly or splenomegaly, increasing volume, induration, erythema and intense pain from the ankle to the popliteal space of her left leg. The urine was dark. Laboratory data were haemoglobin 5 g/dL, hematocrit 17%, reticulocytes 62%, and platelets 281×109/L. Peripheral blood smear showed esquistocytes +++ and spherocytes ++, suggesting intravascular hemolysis. Total bilirrubin 2.99mg/dL, direct bilirubin 0.57, and LDH 750 U/L. Doppler ultrasound: obstruction of the deep and superficial venous system of tibial, peroneal and left popliteal veins. Four red cells units were transfused and FFP transfusion was started every 6 hours, anticoagulation with enoxaparina sodium (1mg/Kg/day) and thrombolysis with rhTPA 100 mg for 3 hours infusion. The patient successfully improved with increase and maintenance of hemoglobin, decrease of the reticulocytes count and evident clinical improvement of her left leg. She was in-hospital for 8 days at the end of which was achieved ambulation, Doppler showed remission of DVT.

The association between exposure to ozone and HIA has not been informed in the absence of G6PD deficiency, and today, little is known of the ideal treatment. Though plasmapheresis is the treatment of choice in a HIA, the presence of DVT and be in a period appropriate for thrombolysis, determined the use of FFP transfusion as the main treatment. The right clinical evolution observed in the treatment of our patient gave her solving clinical problems. Ozone has been widely used for a variety of off-label purposes. In vitro experiments had demonstrated hemolysis with ozone concentration > 30 mcg/mL, therefore this case must represent an important alert for those ozone users, however the mechanism of hemolysis because ozone exposure remains to be elucidated.

Disclosures:

No relevant conflicts of interest to declare.
Serum Uric Acid As a Predictor Factor of the Response to Deferasirox Therapy for Patients with b-Thalassemia Major
Authors: Vlachaki Efthimia , Perifanis Vasileios , Kondou Antonia , Neokleous Nikolaos , Teli Aikaterini , Oikonomou Marina
Published: October 2019

Abstract 5306

Serum uric acid of patients with beta-thalassemia major (b-MA), despite the hyperuricosuria observed in these patients, is usually in the upper normal levels or increased due to excessive catabolism of the red blood cells (ineffective erythropoiesis). Deferasirox a new oral iron chelator with potential nephrotoxicity is recently used as iron overload treatment in patients with b-MA.

Aim:

Aim of the study is to investigate the effect of deferasirox on uric acid levels of patients with homozygous b-MA.

Material-Method:

53 patients were enrolled to the present study with b-MA major, (aged 22.4 ± 14.7 years, range 4–12 years) 36 adults and 17 children, 32 females and 21 male. All the patients were transfusion-dependent with pretransfusional haemoglobin 9gr/dl and treated with iron chelation. The comparison was made between two different time points' measurements of uric acid and ferritin, at the beginning before Deferasirox, and one year later. The blood was taken from the patients early at mornings before the transfusion. Also uric acid was measured in 24 hour urine of patients under deferasirox, or other iron chelation therapy or after weekly discontinuation of deferasirox. Patients taken allopurinol or thiazide or with abnormal kidney function were excluded.

Results:

There is statistically significant difference (p< 0.001) between the mean annual value of serum uric acid (before 5.2 ± 1.3mg/dl) and ferritin (before 1653,4 ± 1026,3 ng/dl) before and after the start of deferasirox (uric acid after 4.2 ± 1.3 mg/dl and ferritin after 1529,07 ± 1137,44 ng/dl). Also, statistically significant positive correlation between the levels of serum uric acid and ferritin was found during the treatment with deferasirox. However, comparing the uric acid in urine of patients, it was not reported any statistically significant difference between treatment with deferasirox (859,75 ± 122), other iron chelators or without iron chelation for one week (844,32 ± 146).

Conclusion:

The mechanism of uric acid reduction in patients with b-MA major treated with deferasirox is not clear. However, it does not seem to be associated with excess of excretion by urine. The positive correlation between uric acid level and ferritin, allow us to consider uric acid as a predictor factor of the response to deferasirox therapy.

Disclosures:

Vlachaki: Novartis Hellas S.A.C.I.: Research Funding. Oikonomou:Novartis Hellas S.A.C.I.: Research Funding.
Von Willebrand Factor (VWF) Regulates Angiogenesis: Studies in Blood-Derived Endothelial Progenitor Cells From Von Willebrand's Disease Patients and VWF-Deficient Mice
Authors: Starke Richard D , Paschalaki Koralia , Ferraro Francesco , McKinnon Thomas A J , Dryden Nicola H , Payne Elspeth , Haskard Dorian O , Hughes Alun D , Cutler Daniel , Laffan Michael , Randi Anna Maria
Published: October 2019

Abstract 5311

Dysregulation of angiogenesis is implicated in many diseases. Von Willebrand factor (VWF), a large plasma glycoprotein essential for normal haemostasis is synthesized by endothelial cells (EC) and megakaryocytes. Raised VWF plasma levels are a risk factor for arterial thrombosis, whilst deficiency of VWF causes Von Willebrand disease (VWD), the most common congenital bleeding disorder in man. VWD can be associated with angiodysplasia, vascular malformations linked to defective angiogenesis which are responsible for intractable bleeding. We recently showed that VWF is involved in angiogenesis. Inhibition of VWF expression in human umbilical vein EC (HUVEC) with specific siRNA resulted in increased in vitro angiogenesis on Matrigel, proliferation and migration. Mechanism studies implicated the endothelial VWF receptor, integrin αvβ3 and the angiogenesis regulator angiopoietin-2. The findings were confirmed in EC from VWD patients and in VWF-deficient mice. Blood outgrowth endothelial cells (BOEC) isolated from peripheral blood of patients with VWD showed decreased VWF release, compared to control BOEC, and increased in vitro angiogenesis, migration and proliferation, similar to what observed with VWF siRNA-treated HUVEC. In vivo studies using the matrigel model and imaging of blood vessels in the ear showed increased angiogenesis and vascular network in VWF-deficient mice compared to controls. Recent studies in patients BOEC and in VWF deficient mice provide new insight into the complexity of this phenotype. We have used the mouse model of post-natal angiogenesis in the retina to carry out detailed analysis of angiogenic networks in the VWF-deficient mouse, and found increased vascular density and defective vascular network. Moreover, using BOEC from patients with type I and type II VWD, we have studied VWF intracellular distribution by immunofluorescence confocal analysis and found patterns of expression that point to a variety of defects in synthesis, storage and secretion. These studies define a new function for VWF, which may have clinical implications for VWD and for patients at risk of CV disease. Moreover, studies with BOEC from VWF patients provide a novel understanding of the physiopathology of this disease.

Disclosures:

No relevant conflicts of interest to declare.
First Case of Platelet-Type Von Willebrand Disease (PT-VWD) Associated with Type 2B Von Willebrand Disease (2B VWD)
Authors: Dreyfus Marie , Desconclois Celine , Guitton Corinne , Baas Marie-Jeanne , Mandard Helene , Boisseau Pierre , Goudemand Jenny , Caron Claudine , Itzhar Nathalie , Veyradier Agnes , Lanza Francois , Fressinaud Edith
Published: October 2019

Abstract 5313

Introduction

VWD 2B and PT-VWD are rare diseases, due to mutations inducing a gain of function respectively of von Willebrand factor (VWF) and of its platelet receptor, Glycoprotein (GP)1bα

Case history

We report here the case of a young girl, born with an extensive purpura and a severe thrombocytopenia: platelet count: 16G/L. There was no associated biological nor clinical abnormality. A high dose of 1g/kg of immunoglobulin G infused on day 1 was unsuccessful, and a HPA-1a (−) platelet concentrate infusion led to a partial and transient increase of the platelet count up to 60G/L. Thrombocytopenia then resolved spontaneously.

Biological study showed no sign of materno-fetal allo- or auto-immunity, parents were not consanguineous.

The diagnosis of type 2B VWD was performed when she was 5 months old: VWF:RCo < 13 IU/dl, VWF:Ag 60 IU/dl, positive ristocetin induced platelet aggregation (RIPA) at a low ristocetin concentration (0.5 mg/ml). RIPA mixing studies were unconclusive.

The same biological abnormalities were found in the father, whereas the mother had normal hemostasis tests. The biological phenotype also included a study of the multimeric VWF structure, showing a marked decrease in percentage of VWF high and intermediate molecular multimers. Genetic analysis performed on VWF gene showed the heterozygous p.Pro1266Leu missense mutation in the VWF A1 domain. This mutation ( o ) is only slightly deleterious, and induces usually a mild disease, without thrombocytopenia, even in stress situations, with normal VWF multimeric distribution; therefore, it could not explain the biological phenotype severity in this family.

GPIBA was then analysed, and a candidate point mutation p.Met239Ile was evidenced. This mutation had not been described yet, but p.Met255Val had already been found in diagnosed cases of PT-VWD.

Conclusion

This case underlines the utmost importance to characterize precisely neonatal thrombocytopenia mechanism. Furthermore, it points out the difficulties to performing PT-VWD diagnosis, which incidence is most probably underestimated. In our case, it was the systematic and extensive biological workout performed in this case of isolated neonatal thrombocytopenia, without any obvious cause, which led to the diagnosis of a PT-VWD, inducing a severe biological phenotype, associated with type 2B VWD characterized by a mild expression. It is, to our knowledge, the first case described to date of such a morbid association.

Disclosures:

No relevant conflicts of interest to declare.
The Phenotype of Endothelium in Children with ALL Might Be a Predictor of the Outcome – a Pilot Study
Authors: Doroszko Adrian , Niedzielska Ewa , Porwolik Julita , Szahidewicz- Krupska Ewa , Chybicka Alicja , Szuba Andrzej
Published: October 2019

Abstract 5314

Background:

Activation of endothelium has been demonstrated to be associated with an increased risk of mortality in severe diseases. In this study we hypothesize that ALL might be also associated with endothelial dysfunction (ED) and that children with concomitant ED might be at higher risk for death.

Methods:

N=18 children at age of 4–18 years with ALL treated with the ALLIC protocol were investigated. Plasma levels of markers of ED (VCAM-1, ICAM-1, E-selectin, P-selectin and PAI-1), lipid peroxidation (malonyldialdehyde – MDA), angiogenesis (VEGF) as well as routine blood tests were analyzed at baseline – the day of diagnosis of ALL, next during the 33rd day of the protocol (steroid therapy, evaluation of remission) and subsequently at the 78th day of therapy (beginning of the protocol M). The control group constituted of N=15, age-matched healthy children.

Results:

Children with ALL were characterized with significantly worse kidney and liver function at baseline as compared to the control. Similarly, the fasting glycemia was higher in the investigated group. Markers of endothelial proagreagattory and procoagulative function (P-selectin and PAI-1) were similar in both groups at baseline. However, vascular inflammation was significantly more pronounced in children with ALL at baseline as compared to the healthy control (ICAM-1= 467.1±72.8 vs. 206.3±21.7 ng/ml, p<0.05). High baseline plasma level of E-selectin and VEGF (as compared to the healthy control) were positive predictors for death in children with ALL (E-selectin: 56.1±10.9 vs. 28.1±10.0 ng/ml, respectively; VEGF: 366.1±61.6 pg/ml vs. 190.1±71.4 pg/ml, respectively, p<0.05).

Conclusion:

Endothelial inflammatory activation is a common phenomenon in children with ALL. High baseline level of VEGF and E-selectin are associated with higher mortality in ALL.

Disclosures:

No relevant conflicts of interest to declare.