bluuebunny/crossref_metadata_2025_split huggingface dataset:
HDAC Inhibitors, FK228 and Valproic Acid, Enhance the Effects of IL-3 or GMCSF on the Generation of Early Megakaryocytic as Well as Erythroid Precursors from Human Hematopoietic Precursors
Authors: Ohishi Kohshi , Ryuu Hyou , Yamamura Kentaro , Masuya Masahiro , Heike Yuji , Takaue Yoichi , Katayama Naoyuki 
Published: April 2020

Abstract
We previously reported that HDAC inhibition by FK228 (depsipeptide) stimulates the proliferation of early erythroid precursors in the presence of IL-3. Here, we examined the effects of HDAC inhibition on human megakaryopoiesis. When G-CSF-mobilized peripheral blood CD34+ cells were incubated in serum-free liquid cultures containing SCF+IL-3 for 8–9 days, cultured cells were subdivided into 4 fractions with the expression of CD61 (GPIIIa) and glycophorin A (GPA). In the semisolid cultures containing SCF+TPO+EPO, CD61+GPA− cell population mainly gave rise to megakaryocytic colonies, while both of erythroid and megakaryocytic colonies were formed from CD61+GPA+ cell population. CD61−GPA+ cells exclusively generated erythroid colonies. After CD34+ cells were cultured with IL-3 or SCF+IL-3 for 8–9 days, addition of FK228 (0.4 ng/ml) to the cultures significantly increased the numbers of CD61+GPA− and CD61+GPA+ cells. Nevertheless, differentiation into CD61+CD42b+ more mature megakaryocytic cells was repressed by FK228. As expected, when cultured cells were replated in the secondary cultures containing SCF+TPO for another 7 days, higher number of CD61+CD42b+ mature megakaryocytic cells were developed in the cultures that had contained FK228 during the initial culture period. Stimulatory effects by FK228 on the generation of early megakaryocytic and erythroid precursors were also seen in the presence of GM-CSF. Moreover, we obtained similar findings with pharmacological concentration (100μg/ml) of valproic acid, which is also class I HDAC inhibitor. These data suggest a modulatory role for HDAC in the IL-3- or GM-CSF-mediated megakaryopoiesis and that the combinatory treatment with HDAC inhibitors and IL-3 or GM-CSF would be useful for stimulating early megakaryopoiesis as well as erythropoiesis in clinical settings.
X-Linked Inhibitor of Apoptosis Protein (XIAP) Supports Urokinase (uPA)-Induced Endothelial Cell Survival
Authors: Prager Gerald W , Mihaly Judit , Brunner Patrick , Zielinski Christoph , Binder Bernd 
Published: April 2020

Abstract
High uPA expressing tumors are associated with poor prognosis. While a direct effect on tumor cell behavior is described, uPA has especially been shown to mediate (tumor-) angiogenesis. Originally, the urokinase system has been implicated to assist the angiogenic process by it’s proteolytic activities. It is now becoming increasingly evident that uPA additionally elicits a whole array pro-angiogenic responses like differentiation, proliferation and cell migration, independent of its proteolytic activity by inducing intracellular signal transduction.
Here we show that uPA induces upregulation of inhibitor of apoptosis proteins (IAPs), which protects endothelial cells against apoptosis. Thereby, uPA-induced endothelial cell survival is mediated by transcriptional upregulation the X-linked inhibitor of apoptosis protein (XIAP), because downregulation of XIAP by small interfering RNA techniques significantly reduces uPA mediated cell survival efficiencies of uPA in endothelial cells. The antiapoptotic activity of uPA was dependent on the presence of uPAR and LRP, but independent of the PI3kinase pathway, while VEGF-dependent antiapoptosis is mainly PI3kinase dependent. uPA-induced cell survival is dependent on the type of extracellular matrix on which cells are attached used indicating the involvement of integrin adhesion receptors. TherebyConsistently, uPA induces phosphorylation of the CDC42 downstream effector p21-activated kinase 1 (PAK1), which leads to IkappaB kinase alpha (IKKa) phosphorylation, a prerequisite for NFkappaB activation. As a consequence, p52/p50 but not p65 is are translocated into the nucleus. Blocking NFkappaB by using the specific NFkappaB inhibitor BAY 11–7082 or by adenoviral-mediated overexpression of its inhibitor, IkB, inhibits uPA-induced XIAP expression as well as uPA-induced cell survival.
From these data we conclude that uPA, which is a main player in endothelial cell migration and invasion, provides an additional, PI3-kinase independent but NFkappaB dependent cell survival mechanism.
Serum VEGF Per Platelet Count as a Prognosis Predicting Factor in Advanced Gastric Cancer Patients
Authors: Kim Byung Soo , Choi Chul Won , Kim Seok Jin 
Published: April 2020

Abstract
Background: New blood vessel formation is a crucial step in the process of tumor growth and systemic metastasis. Recent studies have shown that VEGF expression not in tissues but in serum sample is correlated with tumor vascularity, and the high serum VEGF levels could predict poor prognosis in cancer patients. However there has been no data regarding the clinical and prognostic significance of serum VEGF levels per platelet count in advanced gastric cancer. In this study, we conducted a study to evaluate the prognostic implication of serum VEGF per platelet count in the patients with advanced gastric cancer.
Methods: 111 patients with histologically confirmed gastric cancer, 35 patients with early gastric cancer were included and control serum samples were acquired from 25 healthy volunteers. The levels of VEGF were measured using human VEGF quantitative enzymelinked immunosorbent assay (ELISA). Survival curves were calculated using the Kaplan-Meier method and survival comparisons were made by the log rank test in metastatic gastric cancer. The Cox proportional hazards regression model was utilized for multivariate analyses after univariate analysis defined relevant prognostic variables.
Results: The mean serum VEGF level was higher in the patients of AGC compared to those with EGC and controls (AGC 465 ± 315.8pg/ml; EGC 306 ± 97.8 pg/ml controls 230.8 ± 53.2 pg/ml, P&lt; 0.033). A trend toward a significant positive correlation between serum VEGF and platelet counts was observed in patients of AGC (r = 0.477, P = 0.000, Fig 2) and there was a significant correlation between serum VEGF levels and differentiation of tumor (p = 0.014), stage (p = 0.036). The overall survival (log rank, p =0.0432) and the progression free survival (median 4.5 vs. 8.9 months; log rank, p =0.0116) were significantly shorter in patients with high VEGF per platelet count (≥1.626 pg/106). In the multivarivate analysis, performance status (P=0.025), the presence of peritoneal carcinomatosis (P=0.006), serum VEGF per platelet (P=0.005) were found to be significantly associated with the short progression free survival
Conclusions: This study demonstrated that serum VEGF per platelet count is correlated with short overall survival and progression free survival in advanced gastric cancer patients. Therefore, serum VEGF per platelet may be a useful marker for predicting the prognosis of advanced gastric cancer patients.
Factors Determining Survival after Unrelated Donor Stem Cell Transplantation in Primary Refractory Acute Myeloid Leukemia
Authors: Craddock Charles F. , Labopin Myriam , Finke J. ürgen , Ehninger Gerhard , Bunjes Donald , Kolb Hans-Jochem , Greinix Hildegard , Zander Axel , Polge Emmanuelle , Schmid Christoph , Beelen Dietrich , Rocha Vanderson 
Published: April 2020

Abstract
Allogeneic stem cell transplantation using an HLA identical sibling donor has the capacity to produce long term disease free survival in a significant number of patients with primary refractory acute myeloid leukaemia (AML). The increased availability of volunteer unrelated donors has resulted in allogeneic transplantation becoming a realistic option in large numbers of patients lacking a sibling donor. However the outcome of patients with primary refractory AML after unrelated donor transplantation has not been systematically examined before. We have therefore studied the outcome of 186 adults with primary refractory AML who underwent an unrelated donor transplant from 1995 to 2006. All patients were refractory to induction chemotherapy having received a mean of 2.2 courses of chemotherapy (range 1 to 6). Cytogenetic status at diagnosis is available in 96 patients: 3 patients had good risk cytogenetics, 60 intermediate risk and 33 adverse risk cytogenetics by MRC criteria. The median interval from diagnosis to transplant was 4.2 months (range 2–6 months). 141 patients underwent transplantation from a 6/6 matched unrelated donor and 45 from a mismatched unrelated donor. 150 patients were transplanted using GCSF mobilised peripheral blood stem cells. 136 patients were transplanted using a myeloablative conditioning (MAC) regimen. 50 patients were transplanted using a reduced intensity conditioning (RIC) regimen of whom 30 received a low dose TBI based regimen. 137 patients achieved a complete remission (CR) post-transplant. The 2 year overall survival for the whole group was 31% and 43% for patients who achieved a CR post-transplant. The day 100 non-relapse mortality was 16% (19% for MAC allografts v 9% for RIC allografts). In univariate analysis time to transplant (&lt;4.2 months), presentation white blood cell count, the absence of circulating blasts at the time of transplant, and the use of a RIC regimen were associated with improved survival. Adverse risk cytogenetics at diagnosis was associated with a decreased OS. In multivariate analysis a short time from diagnosis to transplant (p=0.05), the absence of adverse risk cytogenetics (p=0.02) and the use of a RIC regimen (p=0.036) were associated with improved survival. The improved survival in patients receiving a reduced intensity transplant occurred despite their increased age at the time of transplant (54 years v 40 years). On the basis of this study we conclude that there is an important role for unrelated donor transplantation in primary refractory AML-particularly if performed early after diagnosis. The encouraging results obtained using a RIC require further examination.
Epigenetic Manipulation of Cancer Testis Antigen (CTA) Expression: A Strategy for Manipulating the Graft-Versus Leukaemia Response in Patients Allografted for Haematological Malignancies
Authors: Stankovic Tatjana , McLarnon Andrew , Agathanggelou Angelo , Goodyear Oliver , Craddock Charles , Moss Paul 
Published: April 2020

Abstract
An immunologically mediated graft-versus-tumor (GVT) underlies the curative effect of reduced intensity allografts in acute myeloid leukaemia (AML) and other haematologic malignancies. Cancer testis antigens (CTA) represent a family of immunodominant proteins that are variably expressed in haematological malignancies and represent a potential target of a GVT response. Importantly a number of CTA genes demonstrate promoter hypermethylation in solid tumours which can be reversed using demethylating agents such as azacitidine. We have therefore examined whether donor T cell responses to CTAs are observed in patients allografted for AML and multiple myeloma (MM) and whether epigenetic therapies can be used to manipulate T cell mediating killing of haemopoietic targets.
We screened 37 patients with AML and 8 patients with MM, who had not received demethylating agent treatment, for T cell responses to 25 peptides derived from 10 CTA genes, including BAGE, LAGE, MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-C2, RAGE-1, by interferon-γ cytokine secretion assay (IFN-γ CSA). We found CTA specific T cells in 11.1% of patients (5 of 45) with frequencies ranging from 0.0005% to 0.2% (median 0.024%). Subsequently, we studied expression of 15 cancer-testis antigens (CTA) at the RNA and protein levels in AML, MM and Hodgkins’ lymphoma cell lines before and after exposure to demethylating agent 5-aza-2′-deoxycytidine (Aza) and histone deacetylase inhibitor sodium valproate (VPA), as single agents and in combination. We found that expression of CTAs HAGE, SACA3, SPANXB, LAGE, XAGE, MAGEA3 could be induced in a dose dependent manner by Aza alone but not with VPA alone. We also observed that expression induced by Aza treatment was increased further by combination treatment with VPA. Induction of CTAs was confirmed in vitro in primary AML cells and in vivo in 2/6 patients on VPA/Aza trial. Furthermore, using interferon-γ ELISA assay we observed that Aza-induced expression of the CTA MAGEA3 in MM cell lines was accompanied by increased recognition by MAGEA3-specific T cells.
These studies confirm the importance of members of the CTA family as targets of a T cell mediated immune response. Our data demonstrate that the expression of these putative immunodominant antigens in haematological malignancies in disease such as AML and myeloma, can be significantly up-regulated by epigenetic therapies with functional increases in target cell killing and support the use of adjunctive epigenetic therapies after allogeneic transplantation with the aim of augmenting a GVL response. A Phase II clinical trial combining post transplant azacitidine with a reduced intensity allograft is currently ongoing in patients with AML.
MicroRNA Signature in Waldenstrom Macroglobulinemia
Authors: Roccaro Aldo M , Sacco Antonio , Chen Changzhong , Leleu Xavier , Runnels Judith , Azab Feda , Azab Abdel Kareem , Jia Xiaoying , Ngo Hai T , Melhem Molly R. , Burwick Nicholas , Quang Phong , Varticovski Lyuba , Novina Carl D , Rollins Barrett J. , Anderson Kenneth C. , Ghobrial Irene M. 
Published: April 2020

Abstract
Background. MicroRNAs (miRNAs) constitute a class of small, non-coding, 18–24 nucleotide RNAs, that act as negative regulators of gene expression. By repressing several target mRNAs, mature miRNAs play a pivotal role in regulating development, cell differentiation, apoptosis, and cell proliferation. Moreover, miRNAs have been described to play roles in both solid tumors and hematologic malignancies, however the role of miRNAs in WM has not been yet elucidated.
Methods. We performed miRNA expression profiling of bone marrow-derived CD19+ WM cells, compared to their normal cellular counterparts and validated data by qRT-PCR. In vitro and in vivo functional studies were performed on miRNA-155 knockdown WM cells. Effect of miRNA-155 on signaling cascades have been evaluated by western blot and immunofluorescence. NF-kB activity has been investigated using a DNA-binding enzyme-linked immunosorbent assay-based assay. Gene expression profile analysis has been performed on both miRNA-155 knockdown- and control probe-transfected WM cells in order to identify potential miRNA-155 targeted genes. Finally correlation between miRNA signature and prognosis has been evaluated.
Results. We identified a WM specific miRNA signature characterized by increased expression of miRNA-363*/-206/-494/-155/-184/-542-3p; and decreased expression of miRNA-9* (ANOVA;P&lt; 0.01). Our data showed that miRNA-155 regulates proliferation and growth of WM cells in vitro and in vivo, by inhibiting MAPK/ERK, PI3/AKT, and NF-kB signaling pathways. Potential miRNA-155 target genes were identified using gene expression profiling and included genes involved in cell cycle progression, adhesion, and migration. Importantly, increased expression of the 6 miRNAs significantly correlated with a poorer outcome predicted by the International Prognostic Staging System for WM (P&lt;0.01). We further demonstrated that therapeutic agents commonly used in WM (rituximab, perifosine, bortezomib) alter the levels of the major miRNAs identified, by inducing down-modulation of five increased miRNAs (all but miR-206) and upmodulation of patient-downexpressed miR-9* microRNA.
Conclusion. These data indicate that miRNAs play a pivotal role in the biology of WM; represent important prognostic markers; and provide the basis for the development of new miRNA-based targeted therapies in this disease.
Aberrant Overexpressions of MicroRNA-21 and MicroRNA-155 Activate AKT Signaling Via Downregulation of Tumor Suppressors in NK-Cell Lymphoma/Leukemia.
Authors: Tagawa Hiroyuki , Yamanaka Yasuo , Watanabe Atsushi , Takahashi Naoto , Sawada Ken-ichi 
Published: April 2020

Abstract
Abstract 1917
Poster Board I-940

Background:
Natural Killer (NK) cell lymphomas/leukemias are characterized groups of highly aggressive lymphoid malignancies, which are comprised of “extranodal NK/T cell lymphoma, nasal type” and “aggressive NK-cell leukemia”. Notably, these two subtypes show many similarities in their morphologic features, immmunophenotypes and genotypes, and are invariably associated with Epstein-Barr virus (EBV), which suggests they may share the same genetic alterations. The gene(s) responsible for natural killer (NK)-cell lymphoma/leukemia have not been identified.

Materials and Methods:
In the present study, we used Northern and quantitative PCR analyses to screen for and quantitatively assess miRNA expression in NK-cell lymphomas/leukemias using 66 probe sets of miRNAs. For function analysis, we conducted antisense oligonucleotides and lentivirus transfection assays using NK-cell lymphoma cell lines.

Results:
We found that in NK-cell lymphoma lines (n=10) and specimens of primary lymphoma (n=10), levels of miR-21 and miR-155 expression were inversely related, and were significantly higher than those seen in normal natural killer (CD3-CD56+) cells (n=8). To determine the functions of these microRNAs in lymphomagenesis, we examined the effects of antisense oligonucleotides targeting miR-21 (ASO-21) and/or miR-155 (ASO-155) in NK-cell lymphoma lines overexpressing one or both of these miRNAs. Conversely, cells showing little endogenous expression of miR-21 or miR-155 were transduced using lentiviral vectors, leading to their overexpression. Reducing expression of miR-21 or miR-155 led to upregulation of phosphatase and tensin homologue (PTEN), programmed cell death 4 (PDCD4), or Src homology-2 domain-containing inositol 5-phosphatase 1 (SHIP1). ASO-21- and ASO-155-treated cell lines all showed downregulation of phosphorylated AKTser473 (pAKT). Moreover, transduction with either miR-21 or miR-155 led to downregulation of PTEN and PDCD4, or SHIP1 with upregulation of pAKT.

Conclusions:
Because recent reports suggest that EBV infection can lead directly to upregulation of miR-21 and miR-155, aberrant overexpressions of these miRNAs via EBV infection could be the first-hit genetic alterations during NK-cell lymphomagenesis. Collectively, these results provide important new insight into the pathogenesis of NK-cell lymphoma/leukemia and suggest targeting miR-21 and/or miR-155 may represent a useful approach to treating NK-cell lymphoma/leukemia.

Disclosures:
No relevant conflicts of interest to declare.
The Lymphoproliferative Disease of Granular Lymphocytes: Updated Criteria for Diagnosis
Authors: Semenzato Gianpietro , Zambello Renato , Starkebaum Gordon , Oshimi Kazuo , Loughran Thomas P. 
Published: April 2020

AbstractThe lymphoproliferative disease of granular lymphocytes (LDGL), also referred to as LGL leukemia, is a heterogeneous disorder, but is clinically, morphologically, and immunologically distinct. Although LDGL has recently been included in the revised classification of lymphomas as an independent clinical entity, no consensus exists on the criteria to establish the diagnosis. The aim of this report was to refine the parameters needed to make the diagnosis of LDGL. We studied 11 patients with chronic granular lymphocytosis selected from among 195 cases observed by our institutions from three different geographic areas (North America, Europe, and Asia). These cases did not meet the current criteria for inclusion in LDGL, since all patients had less than 2,000 GL/μL. However, in each of these patients, we found evidence for expansion of a discrete GL population. Clonal rearrangement of the T-cell receptor (TCR) β gene was found in peripheral blood mononuclear cells (PBMC) of all nine patients with CD3+ LDGL. Using recently generated monoclonal antibodies (MoAbs) against the TCR Vβ gene regions, we identified a unique TCR Vβ on GL from each of three patients studied. In two patients with CD3− LDGL, we also identified a restricted pattern of reactivity, by staining with MoAbs against p58 antigen found on normal natural killer (NK) cells. The clinical features of these 11 patients with relatively low absolute number of GL were similar to those reported previously for patients with greater than 2,000 GL/μL. These data demonstrate that newer techniques such as MoAbs against Vβ gene regions and p58 molecules and molecular analyses are useful to identify expansions of discrete GL proliferations. Demonstration of an expansion of a restricted GL subset is evidence for the diagnosis of LDGL, even in patients with a relatively low GL count. Our results also contribute to distinguish between the end of normality and the beginning of pathology in the broad spectrum of GL lymphocytoses.
Recombinant Methionyl Human Stem Cell Factor and Filgrastim for Peripheral Blood Progenitor Cell Mobilization and Transplantation in Non-Hodgkin's Lymphoma Patients — Results of a Phase I/II Trial
Authors: Moskowitz Craig H. , Stiff Patrick , Gordon Michael S. , McNiece Ian , Ho Anthony D. , Costa John J. , Broun E. Randolf , Bayer Robert A. , Wyres Melody , Hill Jerome , Jelaca-Maxwell Kathy , Nichols Craig R. , Brown Sherri L. , Nimer Stephen D. , Gabrilove Janice 
Published: April 2020

AbstractTo examine the safety and efficacy of recombinant-methionyl human stem cell factor (r-metHuSCF), 38 patients with intermediate-grade or immunoblastic high-grade non-Hodgkin's lymphoma who were eligible for autologous transplantation were randomized to receive r-metHuSCF (5, 10, 15, or 20 μg/kg/d) plus Filgrastim (10 μg/kg/d) or Filgrastim (10 μg/kg/d) alone to mobilize peripheral blood progenitor cells. Subcutaneous administration of r-metHuSCF was well tolerated in conjunction with a multi-agent pre-medication regimen; local injection site reactions were the most commonly seen adverse event. The total mononuclear cell count, CD34+ cell content, granulocyte-macrophage colony-forming cells (GM-CFC), and burst-forming units-erythroid (BFU-E) per kilogram in the apheresis product was similar when all patients were analyzed by treatment cohort and mobilization regimen (Filgrastim or r-metHuSCF in combination with Filgrastim); however, when prior chemotherapy was taken into account in a supplementary analysis, clinically important differences were observed. Extensive prior therapy was defined as the amount of exposure to specific stem cell toxic chemotherapeutic agents that patients received. These agents include procarbazine, nitrogen mustard, melphalan, nitrosoureas (≥2 cycles of any of these drugs) or greater than 7.5 g of cytosine arabinoside. In these patients, there was an increased number of CD34+ cells (1.76 v 0.28 × 106/kg), GM-CFC (20.5 v 5.0 × 104/kg), and BFU-E (36.9 v 8.9 × 104/kg) in patients receiving r-metHuSCF and Filgrastim (N = 18) compared with Filgrastim alone (N = 5). These patients also had a decreased time to an untransfused platelet count of 20 × 109/L that was 10.5 days shorter in the patients who received r-metHuSCF and Filgrastim (12.5 v 23 days). These differences were not found to be statistically significant, possibly because of small size, but are clinically important.