bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Heparin-Based Inhibitor of Heparanase Blocks Myeloma Growth In Vivo by Targeting the Tumor Microenvironment.
Authors: Yang Yang , Swain Telisha , Naggi Annamaria , Torri Giangiacomo , Casu Benito , Vlodavsky Israel , Sanderson Ralph D. 
Published: April 2020

Abstract
Heparanase is an enzyme that cleaves heparan sulfate chains of proteoglycans and promotes the growth and metastasis of many types of human tumors. Our previous work demonstrates that enzymatically active heparanase
is present in the bone marrow of myeloma patients and is associated with a poor prognosis, substantially enhances tumor growth and spontaneous metastasis to bone in an animal model of myeloma, and increases the synthesis and shedding of syndecan-1 by myeloma cells,
this in turn contributes to myeloma progression by elevating levels of syndecan-1 in the tumor microenvironment. Thus, we hypothesized that inhibitors of heparanase activity would have a dramatic impact on the growth of myeloma tumors. To test this we used a chemically modified form of heparin that is 100% N-acetylated and 25% glycol-split (designated 100NA,RO-H). This form heparin is a potent inhibitor of heparanase enzyme activity but lacks anticoagulant activity thus enabling use of relatively high doses of the drug in vivo. Delivery of the 100NA,RO-H to animals bearing established myeloma tumors dramatically blocked tumor growth and progression in a dose-dependent manner(P&lt;0.04). To understand the mechanism of action of 100NA,RO-H, a series of experiments were performed on tumor tissue harvested from the animals. Results demonstrate that the modified heparin significantly inhibits proliferation of cells within the tumor (33 ± 8/mm2 Ki-67 positive cells in treated vs. 688 ± 164/mm2 positive cells in controls, P = 0.002). 100NA,RO-H also dramatically inhibits angiogenesis as compared to controls as assessed by CD34 staining of tumor tissue. This anti-angiogenic effect may be due at least in part to its regulation of hepatocyte growth factor and/or vascular endothelial growth factor, two angiogenic factors that are detected by immunohistochemistry at high levels in controls but virtually absent from tumors of animals treated with 100NA,RO-H. In vitro studies demonstrate that 100NA,RO-H blocks syndecan-1 shedding from cells, consistent with the role of heparanase in promoting syndecan-1 shedding. In addition, in contrast to our finding that 100NA,RO-H blocks tumor growth in vivo, it only slightly inhibited proliferation, cell cycle progression and growth factor signaling in myeloma cells growing in vitro. Thus, the compound does not appear to have substantial direct effects on tumor cells. Although it is not yet clear if all of the effects of 100NA,RO-H are due to its anti-heparanase activity, we conclude that this modified heparin disrupts the myeloma tumor microenvironment thereby blocking in vivo growth and progression of the cancer. Therefore, its use as a single agent or in combination with agents having direct anti-tumor cell activity may constitute a potent new anti-myeloma therapy.
IRF-4 Functions as a Tumor Suppressor in Early Stages of B Cell Development.
Authors: Acquaviva Jaime , Ren Ruibao 
Published: April 2020

Abstract
Interferon regulatory factor 4 (IRF-4) is a hematopoietic cell restricted transcription factor important for hematopoietic development and immune response regulation. IRF-4 is essential for mature B and T lymphocyte development, dendritic cell differentiation and some macrophage functions. It was also originally identified as the product of a proto-oncogene involved in chromosomal translocations in multiple myeloma. Recent studies show that expression of IRF-4 is essential for the maintenance of multiple myeloma cells, making IRF-4 an attractive target for the development of cancer therapies. In addition to its role in late stages of lymphocyte development, IRF-4 functions redundantly with its closely related family member, IRF-8, to promote differentiation at the pre-B to immature B transition, apparently by repressing the pre-B cell receptor signaling and promoting light chain gene rearrangement. In contrast to its oncogenic properties, expression of IRF-4 is downregulated in BCR/ABL positive acute B-lymphoblastic leukemia (B-ALL) and is increased in response to imatinib, a selective BCR/ABL tyrosine kinase inhibitor. To assess the role of IRF-4 in the pathogenesis of BCR/ABL-positive B-ALL, we examined the effect of both loss of and forced expression of IRF-4 in BCR/ABL leukemogenesis using a mouse bone marrow transduction and transplantation model system. We found that BCR/ABL induced B-ALL significantly faster in IRF-4-deficient cells than control cells. On the other hand, forced expression of IRF-4 potently suppressed BCR/ABL transformation of bone marrow derived B lymphoid precursors in vitro and BCR/ABL induced B-ALL in vivo. These results provide evidence that IRF-4 functions as tumor suppressor in early B cell development and may allow elucidation of new molecular pathways significant to the pathogenesis of BCR/ABL positive B-ALL. In a parallel study, we have shown that IRF-4 functions as a tumor suppressor in myeloid lineage. The context dependent roles of IRF-4 in oncogenesis should raise caution in the development of cancer therapies targeting IRF-4. Further dissecting the mechanisms by which IRF-4 functions as an oncogene or a tumor suppressor gene is crucial for developing therapeutic strategies precisely targeting the IRF-4 oncogenic pathway as well as for developing therapeutic strategies taking advantage of the IRF-4 tumor suppressor pathway.
NF-E2 Overexpression Delays Erythroid Differentiation and Increases Erythrocyte Production
Authors: Mutschler Manuel , Magin Angela S. , Buerge Martina , Will Britta , Pilz Ingo H. , Migliaccio Anna Rita , Pahl Heike L. 
Published: April 2020

Abstract
The transcription factor Nuclear-Factor-Erythroid2 (NF-E2) is overexpressed in the vast majority of patients with polycythemia vera (PV). PV affects precisely those lineages expressing NF-E2: hematopoietic precursors, erythroid, megakaryocytic and granulocytic cells. In murine models, NF-E2 overexpression increases proliferation and promotes cellular viability in the absence of EPO. EPO-independent growth is a hallmark of PV. We therefore hypothesized that NF-E2 overexpression contributes to erythrocytosis, the pathognomonic symptom of PV. Consequently, we investigated the effect of NF-E2 overexpression in healthy CD34+ cells. NF-E2 overexpression lead to a delay in erythroid differentiation, manifested by a belated appearance of GlycophorinA-positive mature erythroid cells. Differentiation delay was similarly observed in primary PV patient erythroid cultures compared to healthy controls. Protracted differentiation lead to a significant increase in the accumulated number of erythroid cells both in PV cultures and in CD34+ cells overexpressing NF-E2. Similarly, NF-E2 overexpression altered erythroid colony formation, leading to an increase in BFU-E formation. These data argue that NF-E2 overexpression delays the early phase of erythroid differentiation, resulting in an expansion of erythroid progenitors, thereby increasing the number of erythrocytes derived from one CD34+ cell. These data propose a role for NF-E2 in mediating the erythrocytosis of PV.
Peripheral Blood Cytogenetic Studies in Myelofibrosis: Overall Yield and Comparison with Bone Marrow Cytogenetic Studies
Authors: Hussein Kebede , Ketterling Rhett P. , Dewald Gordon W. , Van Dyke Daniel L. , Mesa Ruben , Tefferi Ayalew 
Published: April 2020

Abstract
Background: Peripheral blood (PB) is sometimes used in place of bone marrow (BM) for cytogenetic studies in either primary (PMF) or post-polycythemia vera/essential thrombocythemia (post-PV/ET MF) myelofibrosis. In a retrospective study, we examined overall yield from PB cytogenetic studies in MF and where possible compared the results with those obtained by BM cytogenetic analysis.
Methods: Standard laboratory techniques were used to perform cytogenetic studies in both the PB and BM. A successful study constituted the acquisition of at least two analyzable metaphases and an abnormal clone required the presence of a specific cytogenetic abnormality in at least two metaphases. Clinical and laboratory correlations were made using variables collected at the time of cytogenetic studies.
Results I: A total of 242 PB cytogenetic studies were reviewed to identify 59 MF cases; 39 were PMF and 20 post-PV/ET MF. Median age was 60 years (range 36–77) and 37 were male patients. The median duration of the disease at the time of PB cytogenetic studies was 3.5 years (range 0.2–35). At least two analyzable metaphases (median 20, range 2–31) were obtained in 49 (81%) of the 59 study patients; in univariate analysis, this was predicted by presence and degree of circulating myeloid progenitor cells (p &lt; 0.0001), higher PB CD34 count (p &lt; 0.0001), higher leukocyte count (p = 0.0008), and absence of myeloid growth factor therapy (p = 0.005). In multivariable analysis, only the presence of circulating myeloid progenitor cells sustained its significance.
Results II: Twenty two cases had concomitant PB and BM cytogenetic studies performed. Of these, 9 patients had abnormal BM cytogenetic findings; the concomitant PB cytogenetic studies were also abnormal in 5 (56%) patients but normal in the other 4. Conversely, PB cytogenetic studies were abnormal in 12 patients; the concomitant BM cytogenetic studies were abnormal in 5 (41%) and normal in the remaining 7. PB cytogenetic studies were not successful in 4 of the 22 total cases; BM studies were successful in 3 of these 4 cases and the results were normal in all.
Conclusion: In the presence of circulating myeloid progenitor cells, PB can be considered as an alternative to BM for cytogenetic studies in MF patients, especially in view of the difficulty getting BM aspirates in this disease. However, considering the prognostic value of specific cytogenetic abnormalities in MF, the non-trivial discordance between the BM and PB cytogenetic findings, observed in the current study, warrants a prospective evaluation for further clarification.
Platelet vWF, ADAMTS-13, and Neutrophil Activation in Patients with Essential Thrombocythemia (ET)
Authors: Falanga Anna , Marchetti Marina , Russo Laura , Barbui Tiziano 
Published: April 2020

Abstract
ET is a chronic myeloproliferative disorder, the benign clinical course of which can be complicated by both thromboses and hemorrhages. The levels of the largest multimers of plasma vWF, are relevant for the hemostatic balance in this disease. vWF is stored in the cellular granules of both endothelial cells and platelets, from which is released upon activation. Neutrophils and platelets circulate in an activation status in ET patients, leading to an increased formation of platelet/neutrophil aggregates. In this study we evaluated in ET patients, the platelet vWF content and its relation to neutrophil activation. In addition, the activity of ADAMTS13, the enzyme responsible for maintaining the normal size distribution of vWF multimers, was studied. A group of 61 consecutive ET patients (50% carrying the JAK2 V617F mutation) and 30 healthy controls were included into the study. Total vWF antigen (vWF:Ag) and activity (vWF:Act) levels were determined in both plasma and in isolated washed platelets by ELISA and collagen binding assay (CBA), respectively. ADAMTS13 activity was measured in plasma according to Gerritsen’s method. CD11b and leukocyte alkaline phosphatase (LAP) were measured on neutrophil membrane as activation markers. Plasma vWF:Ag levels resulted significantly increased in ET patients compared to controls (p&lt;0.05), the subgroup of patients with the JAK2 mutation showing the highest levels; whereas vWF:Act was not statistically different. The vWF Act/Ag ratio was significantly lower in ET compared to controls (0.85±0.15 vs 0.95±0.23; p&lt;0.01). A significant inverse relation between platelet count and plasma vWF:Act existed. On the other hand, platelets from these patients had a decreased content of both vWF:Ag and vWF:Act than platelets form controls (p&lt;0.01), suggesting an increased platelet release of vWF. In addition, platelets from ET JAK2 mutation carriers had significantly lower vWF:Act compared to wild-type subjects. Accordingly, the platelet vWF Act/Ag ratio of JAK2 mutation carriers was significantly (p&lt;0.01) lower vs wild-type patients and controls. Neutrophil confirmed to be activated in these patients, as previously described. Indeed, surface activation markers were increased in ET patients compared to controls, and, in particular, CD11b levels were inversely related to platelet vWF:Act (p&lt;0.01). The measurement of plasma ADAMTS13 showed a reduction in the activity of this protease in ET patients (68±16%) compared to normal control mean (p&lt;0.01). In conclusion, in ET patients and particularly in JAK2 mutation carriers, a decrease in platelet vWF:Ag and Act is found and correlates with neutrophil activation, suggesting a role of neutrophil in platelet degranulation. Increased vWF release and reduced ADAMTS13 activity can represent a mechanism of hypercoagulation in these patients.
High Skp2 Expression Is an Independent Predictor of Unfavorable Outcome in 333 Patients with Diffuse Large B Cell Lymphoma (DLBCL).
Authors: Seki Ritsuko , Ohshima Koichi , Kawano Fumio , Murayama Toshihiko , Moriuchi Yukiyoshi , Ogawa Ryosuke , Shimoda Kazuya , Suzushima Hitoshi , Tsuda Hiroyuki , Arima Nobuyuki , Tsukazaki Kunihiro , Suzuki Keiko , Iwahashi Masahito , Utsunomiya Atae , Matsumoto Tadashi , Okamura Takashi 
Published: April 2020

Abstract
The addition of rituximab to CHOP (CHOP-R) chemotherapy has resulted in an improved outcome for patients with DLBCL and has recently been shown to diminish the prognostic impact of several recognized biomarkers. S-phase kinase-associated protein 2 (Skp2) is a proto-oncogene that has been shown to be expressed in a number of tumors. We have reported that Skp2 expression in tumor cells is an unfavorable prognostic factor in DLBCL. In the present study, we investigated the significance of Skp2 expression in the patients with DLBCL treated with CHOP or CHOP-R. DLBCL patients (333 cases) were entered into this study, based on the availability of paraffin blocks for interpretable immunohistochemistry for all antigens (CD10, Bcl-6, MUM1, Bcl-2, Skp2). All patients were treated with either CHOP (201) or CHOP-R (132) from 1996 to 2005, and were diagnosed as having DLBCL at the twenty different hospitals. All specimens were histopathologically reviewed before entering into this study. Their clinical characteristics, including either the IPI or R-IPI factors, were evenly matched. The median follow-up of living patients was 3.7 and 2.1 y for CHOP vs CHOP-R, respectively. DLBCL were assigned to GCB subtype (40.8%: 136/333) or non-GCB subtype (59.2%: 197/333) based on the method of Hans et al., Blood 103: 275–82 (2004), with similar distribution in both treatment groups. Expression of bcl-6 (p&lt;0.05) or GCB subtypes (p&lt;0.05) was associated with better overall survival (OS), whereas expression of bcl-2 (p&lt;0.05) was associated with worse OS in CHOP treatment group. The addition of R was associated with an improved survival in the non-GCB subtype and resulted in same as that of GCB subtype. The survival benefit of both low Bcl-2 and high Bcl-6 expressions diminished in combined treatment with R to CHOP. There were 97 patients with high Skp2 expression (&gt;60% positive cells) (97/333: 29.1%). High Skp2 expression was found in both GCB subtype (28.6%) and non-GCB subtype (30.3%). In advanced clinical stage or extranodal involvement (&gt;2), the patients with high Skp2 expression had worse survival than those with low Skp2 expression (p&lt;0.05). Interestingly, in CHOP-R group, high Skp2 expression was the strong biomarker of worse prognosis (p&lt;0.05). DLBCL patients with high Skp2 expression did not benefit from the addition of R to CHOP. Therefore, Skp2 may be a useful prognostic marker in recent rituximab era. The new treatment strategy is necessary for the DLBCL patients with high Skp2 expression.
Figure Figure
Serum B-Cell Activating Factor (BAFF) in Diffuse Large B-Cell Lymphoma: Its Prognostic Significance in the Rituximab Era.
Authors: Kim Seok Jin , Sung Hwa Jung , Choi Chul Won , Kim In Sun , Kim Byung Soo , Yoon Soo-Young , Won Jong-Ho , Kim Jun Suk 
Published: April 2020

Abstract
The introduction of rituximab, a chimeric monoclonal antibody against the CD20 antigen has significantly improved the treatment outcome of diffuse large B-cell lymphoma (DLBCL). However, not all patients with DLBCL respond to rituximab plus CHOP (R-CHOP) chemotherapy. Thus, resistant clones of DLBCL remain a significant clinical problem. Although the underlying mechanisms of this resistance are still not clear, considering that the dysregulation of the balance between cell survival and programmed cell death is a central feature of lymphomagenesis, a factor promoting survival of lymphoma cells may be associated with resistance of DLBCL to rituximab. B-cell activating factor (BAFF), a member of the tumor necrosis factor (TNF) superfamily has shown to be a key mediator in the formation and regulation of normal B-cell response. BAFF is usually expressed by monocytes and macropahges, and is promoting cell survival and preventing apoptosis. Therefore, we performed this study to determine the clinical impact of serum BAFF on the treatment outcome of DLBCL treated with R-CHOP. 48 patients diagnosed as DLBCL were enrolled, and all the patients were treated with R-CHOP every 3 weeks. Before treatment and every two cycles of R-CHOP, their serum was taken and cryopreserved. The level of BAFF was measured by ELSA method (Human BAFF immunoassay, R&D systems, USA). We also analyzed the expression of BAFF receptor in tumor sections by immunohistochemistry (Polyclonal antibody to BAFF receptor, Abcam, UK). Median age of the patients was 56 years old (range 21–79). 20 patients were stage I/II, and 28 were stage III/IV. Rituximab was administered at the dosage of 375mg/m2 on day1, and CHOP chemotherapy was combined with rituximab until 6 – 8 cycles. The mean level of BAFF at the time of diagnosis was 2284.54pg/mL, and the median was 1326.60pg/mL (138.80–13537.92pg/mL). When the patients were dichotomized into high and low BAFF group based on the median value, 21 patients showed complete response in low BAFF group, however, only 3 patients showed complete response in high BAFF group (p=0.011). Among 16 patients showing relapse, 11 patients belonged to high BAFF group. The expression of BAFF receptor was also increased in relapsed patients compared to non-relapse group (p &lt; 0.05). In the analysis of relapse-free survival, low BAFF group did not reach the median value as yet while high BAFF group showed 379 days (p=0.046). Thus, baseline serum BAFF may be associated with response of DLBCL to R-CHOP. When we serially checked the level of serum BAFF, the level of BAFF was increased during treatment (After the 2nd cycle 6448.56pg/mL, and after 4th cycle 7421.28pg/mL). In conclusion, serum B-cell activating factor may be a useful indicator predicting response to R-CHOP and prognosis in patients with DLBCL.
Figure Figure
Signaling Pathways of All-Trans Retinoic Acid (ATRA)-Induced Apoptosis in Mantle Cell Lymphoma (MCL).
Authors: Singh Amareshwar T.K. , Bhalla Savita , Ryan Robert O. , Nguyen Thanh-Son , Singh Karthik T.K. , Evens Andrew M. , Gordon Leo I. 
Published: April 2020

Abstract
MCL, characterized by the t(11;14), results in the overexpression of cyclin D1, and typically has an aggressive clinical course. The disease is not curable by conventional therapy, and new modalities are needed. A MCL cell line, Granta, has been studied and has a complex karyotype and genetic alterations of cell cycle regulatory genes. ATRA is a metabolic derivative of retinoic acid (RA). Upon binding with ATRA, its receptors become transcriptionally activated and transactivate their target genes leading to cell growth arrest or apoptosis. We hypothesized that ATRA would lead to apoptosis or cell growth arrest in Granta MCL cells. We used ATRA nanodisks for increased solubilization and in vivo delivery. Reactive oxygen species (ROS) were measured with fluorescence-activated cell sorting (FACS) following incubation at 6 hours (h) and 24 h with ATRA (12.5–50nM). At 6 h, ATRA induced dose-dependent generation of ROS, which returned to baseline at 24 h. The levels of ROS in untreated controls were unchanged over all the incubation time points. To determine the effects of ATRA on apoptosis, Granta cells were treated with various concentrations for 6 h, 24 h, and 48 h; apoptosis was measured with annexinV/propidium iodide (annexin/PI) binding and FACS analysis. We found increasing apoptosis in Granta at 24 h, reaching significant levels at 48 h. At 48 h, the lowest doses of ATRA (12.5nM) resulted in &gt;70% increase (as a percentage of control) in annexin/PI. To determine the biologic consequences of ATRA exposure in Granta, RARα protein levels were measured in control and ATRA treated cells at 2 h, 6 h, and 24 h. RARα protein was marginally increased at 6 h and significantly downregulated following 24 h ATRA exposure (50nM). Furthermore, ATRA significantly increased p21 protein expression at 24 h as assessed by immunoblotting. RT-PCR data showed that SESN1(T2) (an oxidant gene) transcript was virtually absent in Granta, and that SESN2 and CDKN1A transcripts were not affected by ATRA exposure. In summary, these data demonstrate that ATRA, complexed with nanodisk delivery particles, stimulates ROS generation as early as 6 h in Granta cells. In addition, ATRA induced significant apoptosis at 48 h, downregulated RARα protein, and increased p21 protein. Further studies investigating the role of ATRA in MCL are warranted.