bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bortezomib Could Reduce Cell Viability by Activation of Heme Oxygenase 1 in Multiple Myeloma Cells
Authors: Tibullo Daniele , Giallongo Cesarina , Barbagallo Ignazio , La Cava Piera , Parrinello Nunziatina , Volti Giovanni Li , Palumbo Giuseppe A , Conticello Concetta , Raimondo Francesco Di 
Published: October 2019

Abstract
Abstract 5005
Bortezomib (PS-341, Velcade®) specifically targeting the ubiquitin-proteosome pathway, is the first defined therapeutic proteosoma inhibitor approved by FDA for treatment of Multiple Myeloma (MM). We explored the hypothesis that the activity of Bortezomib in MM cells may involve the heme oxygenase 1 (HMOX1), a enzyme with pleiotropic effects, catalysing the degradation of heme to carbon monoxide, biliverdin and ferrous iron.
After 24h of treatment with Bortezomib at 10nM, the U266 and SKM-M1 MM cell lines showed a decrease of cell viability around 50% and an increase of HMOX1 protein and mRNA expression of 40 fold (p<0. 001). Using a colorimetric assay we observed that Bortezomib increased also the HMOX1enzymatic activity (p<0. 03). To elucidate the significance of the HMOX1 increase after bortezomib treatment, we treated myeloma cells with Bortezomib plus an inducer (Hemin 10uM) or an inhibitor (Tin, Sn-Mesoprotoporphyrin, 10uM) of the enzyme activity. Treatment with Hemin and Tin alone not changed the cell viability. Cell viability was reduced (p<0. 003) by addition of Hemin and was increased (p<0. 002) by Tin addiction with respect to Bortezomib alone.

Conclusion:
Bortezomib increases HMOX1expression and its enzymatic activity and this increase could be an important mechanism of Bortezomib induced cytotoxicity in MM cells. Modulation of HMOX1 activity has effect on Bortezomib induced cell death.

Disclosures:
Off Label Use: Eltrombopag is a thrombopoietin receptor agonist indicated for the treatment of thrombocytopenia in patients with chronic immune (idiopathic) thrombocytopenic purpura (ITP).
Cytogenetic Analysis and Correlation Study in Prognosis of Patients with Multiple Myeloma
Authors: Fu Chengcheng , Xin Xue , Depei Wu 
Published: October 2019

Abstract
Abstract 5006
Multiple myeloma (MM) is a fatal neoplasm characterized by the accumulation of malignant plasma cells within the bone marrow (BM) and the presence of a monoclonal immunoglobulin in the serum and/or urine, it was reported that patients with abnormal metaphases by conventional cytogenetics at diagnosis had active disease and a reduced survival rate compared with those normal metaphase. To further explore the cytogenetic characteristics of multiple myeloma and its correlation with clinical treatment and prognosis, 38 cases of MM were collected and chromosome specimens of bone marrow cells were prepared by 24-hour culture. R-banding technique combined with interphase fluorescence in situ hybridization(i-FISH) was used for karyotype analysis. The impact of different treatments and chromosome aberrations on progression-free survival(PFS) and overall survival(OS) time as well as related prognosis factors of MM were analyzed.
The detected total chromosome aberration rate was 34. 2% (13/38), with chromosome complex aberration 53. 8%(7/13). The detection rate of R-banding technique was 23. 7%(9/38) and 21. 1%(8/38) by FISH. The most frequent chromosome aberration was chromosome 1 abnormality. The median PFS time of abnormal group was 16 months and was not reached of normal group after median follow up for 36 months(P=0. 045). Also, the median PFS time of complex abnormal group was 15 months, and the median PFS time of non-complex abnormal group was not reached(P=0. 012). The median OS time of complex abnormal group was 22 months and the median OS time of non-complex abnormal group was not reached(P=0. 041). Boretizomib or autologous hematopoietic stem cell transplantation prolonged the PFS and OS of the chromosome aberration group compared with traditional chemotherapy, but there were no significant statistical differences between the two groups(P>0. 05).
It was concluded that chromosome complex aberration is the mainly cytogenetic characteristics of multiple myeloma, and multiple numerous and structural aberrations are involved. The patients with chromosome complex aberration have poor prognosis. Chromosome abnormalities detected in metaphases from multiple myeloma (MM) cells have a clear impact on prognosis and response to therapy. Techniques to explore cytogenetic and molecular genetic changes of myeloma cells should be used to improve chromosomal aberration detection to guide clinical treatments and evaluate prognosis.

Disclosures:
No relevant conflicts of interest to declare.
Inhibition of Related JAK/STAT Pathways with Molecular Targeted Drugs Shows Strong Synergy with Ruxolitinib in Chronic Myeloproliferative Neoplasms
Authors: Barrio Santiago , Gallardo Miguel , Arenas Alicia , Ayala Rosa M. , Rapado Inmaculada , Rueda Daniel , Jimenez Ana , Albizua Enriqueta , Burgaleta Carmen , Gilsanz Florinda , Martinez Joaquin 
Published: October 2019

Abstract
Abstract 5054

Objectives:
In the present study we have determined the antitumor effects, molecular mechanisms of action and potential synergies between Ruxolitinib and the molecular targeted drugs Sorafenib, KNK437, Dasatinib and Perifosine, in Philadelphia negative chronic myeloproliferative neoplasms (MPN).

Materials and methods:
The cytotoxic and cytostatic effects of the different compounds were determined in the JAK2V617F positive cell lines HEL and Ba/F3 JAK2V617F EPOR, and in mononuclear and bone marrow CD34 positive cells from 19 MPN patients. The effects of different drugs at the molecular level were analyzed by flow cytometry and western blot. The IC50 and the synergy combination index (CI) were determined with GrafPath Prism software and Calcusyn software respectively.

Results:
Ruxolitinib (IC50PV=15nM), as well as Sorafenib (IC50PV=8uM), KNK437 (IC50PV=100uM) and Perifosine (IC50PV=15uM), was able to inhibit proliferation in cell line models and in cells from MPN patients. Dasatinib (IC50PV=12nM), however, was only effective in patient samples, with similar effects in both MPN patients and control donors. Moreover, Dasatinib, KNK437 and Sorafenib showed a strong synergistic effect in combination with Ruxolitinib (CIPV<0. 3, table 1). The Western blot analysis confirmed that Sorafenib inhibited the activation of ERK and P38, and, as a consequence, of STAT5. Ruxolitinib blocked the ERK and STAT5 activation, Dasatinib blocked SRC and STAT5, and KNK437 decreased the stability of the protein JAK2, reducing its expression.

Conclusions:
The blockage of JAK2 related proliferative pathways has the potential to inhibit cell proliferation in MPNs. Furthermore, the combination of Ruxolitinib with inhibitors targeted against these pathways has a strong synergistic effect. This may be related to a decrease in the activation of the final common effector STAT5.

Disclosures:
No relevant conflicts of interest to declare.
Sezary Syndrome: Clinicopathological and Immunophenotypical Characteristics and Their Impact On Survival
Authors: Deaver Darcie , Horna Pedro , Sokol Lubomir 
Published: October 2019

Abstract
Abstract 5085

Background:
Sezary syndrome is a rare leukemic subtype of cutaneous T cell lymphoma with aggressive clinical behavior and poor prognosis compare to the most common type of CTCL, mycosis fungoides. Hematologic criteria for the diagnosis of Sezary syndrome include an absolute Sezary cell count of >1000cells/mm3 in peripheral blood the presence of a clonal T cell population, increased CD4/CD8 ratio, and aberrant expression of T cell markers.

Objectives:
To evaluate clinical, pathological and immunophenotypical characteristic in patients with Sezary syndrome and determine the impact of these factors on disease survival.

Research Design and Methods:
Retrospective chart review of 17 consecutive patients diagnosed with Sezary syndrome at Moffitt Cancer Center from January 1998-June 2012. Data points collected were age, gender, stage of disease, date of diagnosis, date of death, and analysis of peripheral blood by flow cytometry. Statistical analyses were performed by SPSS statistical software (IBM, Somers, NY). All causes of death were included in the survival analysis. Survival curves were estimated by the Kaplan-Meier method.

Results:
Of the 17 patients evaluated 10 (58. 8%) were female, and 7 (41. 2%) were male, median age was 71 years (range 50–89). Patients were staged according to 2007 ISCL/EORTC staging system 8 (47. 1%) stage III and 9 (52. 9%) stage IV. Median survival was 20. 9 months (range 0–107. 6) for all stages; 27 months and 9. 36 months for stage III and IV, respectively. As expected, stage IV disease was associated with a higher CD4:CD8 ratio (median 62. 5/uL versus 8. 6/uL), absolute lymphocyte count (median 9, 860/ul versus 2800), and absolute Sezary cell count (median of 8, 700/uL versus 1, 700/uL). Interestingly, stage IV disease was also associated with a higher absolute NK cell count (median 280/uL versus 60/uL) and a lower absolute CD8 T cell count (median 165/uL versus 290/uL). Of all parameters studied the only ones showing statistical correlation with the stage of disease were the NK cell count (p = 0. 018) and the CD4:CD8 ratio (p = 0. 039). The immunophenotype of the Sezary cells did not correlate with stage of disease. Survival analysis did not show any significant differences by grouping patients according to high or low levels of variables described above, although the series is small.

Conclusion:
We concluded that increased burden of disease in the peripheral blood did not affect overall survival in our patient population. However, Higher CD4:CD8 ratio, higher absolute lymphocyte count, and lower absolute CD8+ T cell count was associated with more advanced stage of disease suggesting that a lack of cytotoxic T cells can be responsible for profound immunosuppression and disease progression found in patients with advanced stage of CTCL.

Disclosures:
No relevant conflicts of interest to declare.
The Incidence, Characteristics, and Outcomes of Venous Thromboembolism in Patients with Follicular Lymphoma Compared to Diffuse Large B-Cell Lymphoma
Authors: Wang Tzu-Fei , Liu Weijian , Carson Kenneth R. 
Published: October 2019

Abstract
Abstract 5090

Background:
Previous studies have demonstrated a widely variable incidence of thromboembolism (VTE) in non-Hodgkin lymphoma (NHL) patients, ranging from 3% to 60%. This is in part because many previous studies have considered NHL as a single disease entity, combining a heterogeneous group of histological subtypes, each with a potentially different thrombogenic potential. We sought to clarify differences in VTE incidence between the indolent and intermediate grade forms of NHL, by looking at follicular lymphoma (FL) and diffuse large B-cell lymphoma (DLBCL), respectively. Furthermore, since chemotherapy is a major risk factor for VTE, we compared VTE incidence rate in the pre- and post-chemotherapy time periods.

Patients/Methods:
We assembled a retrospective cohort of the United States veterans diagnosed with DLBCL or FL between 1998 and 2009. VTE was identified by ICD-9 codes, while DLBCL or FL was identified by ICD-O3 codes in the Veterans Health Administration central cancer registry. Patients who did not receive any chemotherapy or had a history of VTE prior to the diagnosis of lymphoma were excluded. VTE rate was compared by Chi-square testing.

Results:
A total of 2094 DLBCL patients and 470 FL patients were included. In the period between lymphoma diagnosis and treatment initiation, VTE occurred at a rate of 18. 0 per 100 person-years in DLBCL and 2. 3 per 100 person-years in FL (p&lt;0. 0001). Similarly, in the 6 months following treatment initiation, DLBCL patients had a VTE rate of 12. 3 per 100 person-years, compared to 7. 3 per 100 person-years in FL (p=0. 039). In DLBCL, patients had a higher risk of VTE between diagnosis and treatment compared to the 6 months after treatment initiation (p=0. 038). In FL, treatment was associated with a trend toward higher risk of VTE, although not statistically significant (p=0. 056).

Conclusions:
DLBCL is more “thrombogenic” than FL both before and during treatment. To our knowledge, this is the first report of direct comparison of VTE event rates between different histological subtypes of NHL, knowledge of which could influence decisions related to thromboprophylaxis. There are limitations of ICD-9 codes which should be acknowledged, and further research will be required to identify the optimal method to identify patients with VTE. The lower incidence of VTE after treatment initiation in DLBCL patients suggests that the reduced disease burden associated with treatment may actually decrease the overall risk of VTE despite the increased risk associated with chemotherapy. Conversely, in FL the risk of VTE appears to be higher in association with treatment.

Disclosures:
No relevant conflicts of interest to declare.
Immunohistochemical Expression of TCRβ Predicts Achievment of Complete Remission in Enteropathy-Associated T-Cell Lymphoma
Authors: Czapiewski Piotr , Fisch Paul , Marks Reinhard , Finke Juergen , Schmitt-Graff Annette 
Published: October 2019

Abstract
Abstract 5101

Introduction:
Enteropathy associated T-cell Lymphoma(EATL) is a distinctive clinicopathological entity but knowledge about potential prognostic factors in this disease is very limited.

Aim:
Clinicopathological and immunohistochemical analysis of single center EATL cohort focusing especially on stromal components, activation molecules and potential loss of TCR receptors.

Methods:
Detailed clinical analysis of a cohort of 21 patients from the southern-west Germany was performed. Additionally broad panel of immunohistochemical staining including MUM1IRF4, Blimp-1, FoxP3, TCR γ, TCRβ, CD11c was done. Univariate statistical analysis of survival was performed.

Results:
The study group comprised of 12 males and 9 females, aged 37–86, with the median survival time of 5 months after the initial diagnosis. Following clinical factors turned out to be connected with longer overall survival (OS): age ≤ 60years (p=0, 007), Ann Arbor Stadium I+II (p =0, 048), good general condition (p = 0, 0068), IPI 1–3 (p=0, 004), usage of anthracycline in the chemotherapy regimen (p=0, 0504).
Longer event free survival(EFS) was connected with good general condition (p=0, 023) usage of anthracycline (p=0, 047).
There was 17 Type I cases (81%) and 4 Type II (19%). EATL type, histological variants of neoplastic cells, perivascular and angiocentric growth pattern as well as tissue eosinophilia did not influence survival. There was a strikingly strong infiltration of the neoplastic infiltrate by CD11c+ dendritic cells in 9/21 cases, but it was prognostically irrelevant. MUM1/IRF4 expression was observed in 8/19(42, 1%), Blimp-1 in 5/14 (35, 7%) and FoxP3 in 7/20 (35%), but only FoxP3 positivity was connected with longer EFS. There was a trend toward worse OS (p=0, 18) and EFS (p=0, 12) in Blimp-1 positive cases.
Expression of TCRβ was observed in 4/21(19%), of TCRγ in 3/21(14, 3%) while the majority of cases (14/21), showed loss of expression of both proteins. There was a trend toward longer OS (p=0, 19) and EFS (p=0, 16) in TCR β+ than in TCRγ+ and TCR γ-β- cases.
TCRβ expression was statistically significant more frequent in patients that achieved complete remission (3/7) than in group that did not achieve CR (1/14, p=0, 049).

Conclusions:
In EATL several clinical factors can provide relevant prognostic information. Immunohistochemical expression of FoxP3, Blimp-1 and TCRβ seems also to be prognostically usefull. TCRβ is the only marker that can predict response to the chemotherapy in this very aggressive neoplasm.

Disclosures:
Finke: Fresenius Biotech GmbH: Honoraria, Research Funding.
Detection of SNPs of MTHFR Genes in Five Hematological Malignant Tumor Cell Lines
Authors: Zhang Xiaoping , Jian Ziying , Chen Baoan , Xu Peipei , Peng Miaoxin , Xia Guohua , Cheng Lu , Lu Zuhong 
Published: October 2019

Abstract
Abstract 5130

Background & Aim:
This study detected three single nucleotide polymorphisms (SNPs) of methylenetetrahydrofolate reductase (MTHFR) gene in five hematology malignant cell lines, to find correlations between MTHFR gene SNPs and hematological malignancies by horizontal comparison.

Materials & Methods:
Cell line K562, Ka, HL-60, U937, and Raji were cultured. The genomic DNA was isolated by QIAamp DNA Blood Mini kit. Designing primes were amplified by PCR to get the related DNA fragments. MTHFR gene A1298C (rs1801131), C677T (rs1801131), G1793A (rs2274976) was genotyped by means of matrix assisted laser desorption ionisation-time of flight mass spectrometry method (MALDI-TOFMS).

Results:
The results showed that the genotype of locus A1298C (rs1801131) on MTHFR gene was AA for U937 cell line. While for K562, Ka, HL-60, and Raji cell lines, that was CA. For all the five hematology malignancies cell lines, the genotype of locus C677T (rs1801133) on the MTHFR gene was CC, and that of locus G1793A (rs2274976) was GG.

Conclusion:
The genotype of locus A1298C (rs1801131) on MTHFR gene was not all the same in the detected hematology malignant cell lines, while the genotypes of the other 2 loci were same in the 5 cell lines.

Disclosures:
No relevant conflicts of interest to declare.
Rituximab Is an Effective Therapy for Relapsed Thrombotic Thrombocytopenia Purpura
Authors: Jajeh Ahmad 
Published: October 2019

Abstract
Abstract 5139
Rituximab, a monoclonal antibodies designed against a CD20 glycoprotein expressing lymphocytes. This drug has been implemented in the treatment of B Cell Lymphoproliferative disorders as well as other non-neoplastic diseases such as autoimmune disorders. Rituximab has produced response in auto-immune thrombocytopenia, auto-immune hemolytic anemia and refractory inhibitors to coagulation factors. The proposed mechanism is not yet well known but it is postulated that rituximab affect the balance between the cytotoxic T cells and B cells. This abstract present our experience with rituximab in the treatment of relapsed Thrombotic Thrombocytopenia Purpura and Hemolytic Uremic Syndrome TTP/HUS. a total of nine patients diagnosed with relapsed TTP/HUS with mean age of forty five years; range 27– 59. Eight black females and one white male with median time to relapse of 3 months (range from 3 weeks to two years). All patients were treated initialy with plasmapheresis and steroids and achieved complete response. All maintained on tapering course of steroids for ten weeks. Upon relapse patient were given rituximab 375 mg/M2 weekly; range 3–6 weeks. This was given after treatment with plasmaphersis/steroids; range 9–25 sessions. Peripheral blood smears, LDH, CBC, BUN and creatine were monitored. In most patients schistocytes were reduced to three per 5000 red cells.
In conclusion: Rituximab is an effective therapy in relapsedTTP/HUS, larger study needed to confirm efficacy. Rituximab to be considered in the first line therapy as well as maintenance is a valid point.

Disclosures:
No relevant conflicts of interest to declare.
RHD*Weak D Type 38: A Family Study
Authors: de Menezes Costa Sidneia Sanches , Chiba Akemi Kuroda , Martin Fabio Oliveira , Langhi Dante Mario , Chiattone Carlos , Bordin Jose Orlando 
Published: October 2019

Abstract
Abstract 5155

Introduction:
The RHD*weak D type 38 is produced by the single nucleotide change (833G>A) in RHD exon 6, carrying a Gly278Asp substitution in the transmembraneous RhD protein. This RHD allele shows a reduced expression of the D antigen on the red blood cell (RBC) surface and may be erroneously typed as D− by standard serologic methods, including the indirect antiglobulin test (IAT). The frequency of the RHD*weak D type 38 in Caucasians is reported to be 1. 5% in Ccee phenotype, while we have recently detected a relatively higher frequency of 2. 6% among Ccee phenotype in Brazil. In this study we performed a serologic and molecular analysis of three generations from a family with the rare RHD*weak D type 38.

Study design and methods:
The RBCs from 45 years-old woman blood donor tested D negative by routine serologic tests (Ccee phenotype) but were positive by the adsorption-elution technique. Further serologic investigation to establish the patterns of reactivity of the RBCs was preformed using anti-D Moabs IgM (clones HM10, P3X63, P3X21211F1, P3X21223B10; Diagast, Loos, France) and anti-D Moabs IgG/IAT (clones HM16, P3X35, P3X241, P3X249, P3X290; Diagast, Loos, France, and MS-26, ESD1; DiaMed, Latino América S. A.). Flow cytometric methods were employed to evaluate the D antigen densities, and molecular sequencing studies were done using a sequencing Kit (Big Dye Terminator v1. 1, applied Biosystems, Weiterstadt, Germany) and a genetic analyser (ABI 3100, Applied Biosystems, Foster City, CA, USA).

Results:
We found that the RBCs reacted negatively with 4/4 IgM anti-D, but showed weakly positive results with 4/7 IgG anti-D reagents. In face of such results we studied 12 members of her family and found that 6 more individuals tested D negative by standard serologic tests (Ccee phenotype) but were positive by adsorption-elution technique. The flow cytometry investigation of samples from the 7 subjects showed D antigen density in a range of 60 – 80 sites per cell and a Rhesus Index of 0. 1 – 0. 6. DNA sequencing analysis from 7 family members was performed showing that all descendants (siblings) had the same RHD*weak D type 38 genotype as the mother, also inherited by the grandson.

Conclusions:
The molecular analysis identified the presence of RHD*weak D type 38 in the three generations of a Brazilian family. The study revealed maternal inheritance (first generation) of the weak D type 38, with Ccee phenotype by her six children and the second generation showed paternal inheritance in a son. We also emphasize the significant prevalence of RHD*weak D type 38 in the Brazilian population, often showing a low antigen D density.

Disclosures:
No relevant conflicts of interest to declare.