bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reduced Intensity Allogeneic Stem Cell Transplantation for Hodgkin’s Disease. Outcome Depends Primarily on Disease Status at the Time of Transplantation.
Authors: Robinson Stephen P. , Schmitz Norbert , Taghipour Goli , Sureda Anna 
Published: October 2019

Abstract
The use of reduced intensity conditioning regimens has extended the application of allogeneic stem cell transplantation to patients previously considered to old or medically unfit for high dose chemo-radiotherapy. However, the role of reduced intensity allogeneic transplantation (RIT) in the treatment of Hodgkin’s disease remains controversial due in part to the limited information available on the outcome of this procedure.We have therefore analysed a large cohort of patients with HD who have undergone RIT. A total of 311 patients from 127 centres were reported to the EBMT registry. Their median age at transplantation was 31.3 years (range 8–61) and 57% were male. They had undergone a median of 2 lines of prior therapy (range 1–6) and 45% of patients had undergone a prior high dose procedure. The median time from diagnosis to RIT was 3.2 years (range 0.25–24.5 years). At transplantation 158 patients had chemosensitive disease, 100 had chemoresistant disease and 53 had untested relapse. All patients underwent conditioning with reduced intensity regimens followed by bone marrow stem cell transplantation from matched family donors (221), matched unrelated donors (61) or mismatched donors (17). Peripheral blood stem cells were used in 263 patients, bone marrow in 46 and a combination in 2. In 294 cases assessable for engraftment sustained engraftment was seen in 93.6%. Chimerism studies performed in 115 patients revealed full donor chimerism in 85 and mixed chimerism in 30. Acute graft versus host disease (grade II–IV) was reported in 24% of patients and chronic graft versus host disease in 20%. With a median follow up of 1 year 59% of patients remain alive and the projected 2 year overall survival (OS) was 46%. In a univariate analysis only disease status at transplantation predicted for a worse OS (p<0.0001). The 100 day transplant related mortality (TRM) was 17% but this increased to 24% at 1 year and 27% at two years and was significantly worse for patients with chemoresistant disease at transplantation (p=0.02). Patient age and prior high dose therapy had no significant impact on TRM. At 1 and 2 years following transplantion 48% and 64% of patients had experienced relapse or progression of their disease. Only disease status at transplantation predicted for a higher risk of disease progression. Consequently the progression free survival (PFS) at 2 years was 26% and was significantly worse for patients with chemoresistant disease (p<0.0001). In univariate analysis patient age, prior high dose therapy and number of lines of prior therapy had no significant effect on PFS. Patients with HD may undergo RIT with acceptable toxicity irrespective of age and prior high dose therapy. However, the outcome depends upon the chemosensitivity of the disease at the time of transplantation and this factor should be a major consideration in the selection of patients for RIT.
Phenotypic and Molecular Characterization of Bone Marrow L-NGFR + Mesenchymal Stem Cells.
Authors: Quirici Nadia , Soligo Davide , Borsotti Chiara , Scavullo Cinzia , Zangrossi Stefano , Lambertenghi-Deliliers Giorgio 
Published: October 2019

Abstract
In a previous report we demonstrated that the immunomagnetic sorting of bone marrow (BM) cells labeled with low-affinity nerve growth factor receptor (L-NGFR) antibodies allows the selection of phenotypically and functionally homogeneous cells that are capable of expansion, self-renewal and differentiation into multiple mesenchymal cells lineages. Furthermore, we reported the presence of a subpopulation of L-NGFR+ cells coexpressing CD133 and CD34, markers associated with a primitive hematopoietic stem cell phenotype. In the present study we expanded on the phenotypic characterization of these cells and investigated their potential for multilineage differentiation.
BM L-NGFR+ cells were analyzed by flow cytometry immediately after immunoseparation and the expression of a variety of stem cell markers was studied. In 12 subsequent experiments L-NGFR+ cells expressed CD45low (97.5% ±3), CD34 (19.9%±13), CD133 (10.4%±6), CD105 (46.8%±36%), P1H12 (50.5%±18), KDR (34%±18) and SSEA-3 (0.47%±0.41). In addition L-NGFR+ expressed high levels of the SCF ligand CD117 (40%±16%). As we previously demonstrated, L-NGFR antibodies identify a subpopulation of cells with a high proliferative capacity and potential for multilineage differentiation along the mesenchymal lineage. We now show, in accordance to these phenotypic data, that the L-NGFR+ cells in the presence of SCF (100 ng/ml) doubled the number of CFU-F and expanded both adipocytic and osteoblastic differentiation in comparison to mesenchymal cultures without growth factors or supplemented with Flt-3L+IL-6 (both 100 ng/ml). SCF seems therefore to act at least as a survival/proliferation factor for mesenchymal stem cells.
Transdifferentiation potentialities towards endothelium were determined incubating L-NGFR+ cells in M199 supplemented with 10% FBS, 50 ng/ml VEGF, 1 ng/ml bFGF and 2 ng/ml IGF-1. At confluence, the cells were further purified using Ulex europaeus agglutinin-1 (UEA-1)-fluorescein isothiocyanate (FITC) and anti-FITC microbeads and expanded with VEGF. Immunophenotypic analysis of 8 samples showed a variable expression of endothelial markers: P1H12 ranging from 17 to 58%, CD105 from 98–100% and CD202b from 18 to 100%.
L-NGFR+ cells, immediately after immunoseparation, were expressing Desmin but not MyoD, Miogenin, Mrf4, Myf5 by means of RT-PCR, while these cells were expressing NSE, TRKA and GalC, but not Nestin and GFAP. Experiments are ongoing to demonstrate muscle and neuron-glial differentiation in vitro using specific media (DMEM 10% FBS + 3 mM %-azacytidine, astrocyte conditioned media, neural stem cell conditioned media),
In conclusion, the expression on NGFR+ cells of a variety of markers, not exclusively related to the mesenchymal lineage, and the reproducible ability to differentiate endothelial cells suggest that these cells may represent a subset of adult MSC with some multipotentiality.
Role of the Hypoxic Bone Marrow Microenvironment in Multiple Myeloma Tumor Progression.
Authors: Asosingh Kewal , De Raeve Hendrik , de Ridder Mark , Storme Guy A. , Willems Angelo , Van Riet Ivan , Van Camp Benjamin , Vanderkerken Karin 
Published: October 2019

Abstract
Recently we reported that pre-clinical myeloma disease progression in the 5T2MM mouse model is characterized by predominant CD45+ MM-cells in the early, pre-angiogenic stage stage of slow tumor progression, followed by expansion of CD45− MM-cells during the subsequent angiogenic stage of progressive tumor growth. Unlike other cancer cells, multiple myeloma (MM) cells have to survive and to grow in a microenvironment which is already hypoxic by nature. This hypoxic bone marrow (BM) microenvironment is essential for normal hematopoiesis. However, the role of BM hypoxia in myeloma tumor progression is not known. Herein we addressed this topic in the 5T2MM mouse model.
Flow cytometric analysis of control mice and 5T2MM diseased mice injected with pimonidazole hypoxyprobe indicated that both normal BM and myeloma infiltrated BM are hypoxic. However, in myelomatous BM the hypoxia was significantly decreased. Analysis of HIF-1a expression, a surrogate marker of hypoxia, by flow cytometry also demonstrated significantly lower levels of hypoxia in myeloma infiltrated BM. HIF-1a expression was found in 5T2MM-cells and was significantly higher compared to the non-tumor cell fraction. In vitro culturing of 5T2MM cells under hypoxic conditions, indicated increased activation of apoptosis inducing caspase-3 in the CD45− MM-fraction, but not in the CD45+ 5T2MM-cells, suggesting that native BM hypoxia selects the tumor population for tumor initiating CD45+ 5T2MM-cells. Although angiogeneic switch and angiogeneic heterogeneity has been reported in MM, the role of myeloma associated angiogensis is remains unclear. The decreased hypoxia in myeloma infiltrated BM adds strength to the hypothesis that myeloma associated neovascularization is functional by increasing BM oxygenation. The data also suggest that the angiogenesis allows expansion of CD45− 5T2MM-cells by decreasing BM hypoxia.
All together, these findings suggest an important role of BM hypoxia in myeloma tumor progression.
Endothelial Cells Induce Multiple Myeloma Cell Proliferation Protect Against Conventional and Novel Therapies.
Authors: Kumar Shaji , Raje Noopur , Hideshima Teru , Podar Klaus , Ishitsuka Kenji , Rocarro Aldo , Chauhan Dharminder , Munshi Nikhil C. , Anderson Kenneth C. 
Published: October 2019

Abstract
Angiogenesis or formation of new blood vessels from existing blood vessels, in contrast to vasculogenesis or de novo formation of new vessels, plays an important role in the progression and spread of most cancers. Multiple myeloma (MM) is characterized by increased microvessel density (MVD), a quantitative estimate of angiogenesis, which correlates with stage of disease. MVD increases with progression from MGUS to smoldering MM to newly diagnosed MM and relapsed MM. It is a powerful prognostic factor, predicting for overall survival. To further elucidate the biological basis for the prognostic value of increased angiogenesis in MM, we studied the interactions of MM cells with endothelial cells using HUVECS as a model system. Co-culture of MM cells (MM1.S, OPM2, U266) with HUVECS induced tumor cell proliferation. Enhanced tumor cell proliferation correlated with the number of HUVECs and was greater than that triggered by co-culture with patient bone marrow stromal cells. When HUVECs were fixed prior to co-culture there was a significant decrease in the tumor cell proliferation. Addition of HUVEC conditioned media to the MM cell lines also induced proliferation. Importantly, HUVECS protected against anti-MM agents including conventional agents (Dexamethasone, Doxorubicin, Melphalan) and novel drugs (Revlimid™). The protective effect afforded by co-culture was lost on HUVEC fixation. Intracellular signaling events following MM cell-endothelial cell contact were studied to understand the mechanisms of the proliferative and protective effects. Western blotting demonstrated activation of the JAK/STAT, PI3K/Akt and the MAPK pathways, mediating proliferation and survival. Ongoing studies focused on understanding cytokine as well as adhesion-mediated interactions between the endothelial cells and the MM cells will identify targets for new therapeutic approaches in MM.
Inhibition of Multiple Myeloma Cell Adhesion to Fibronectin by Ephrin Ligation.
Authors: Ratner Stuart , Schiffer Charles A. , Zonder Jeffrey A. 
Published: October 2019

Abstract
Multiple myeloma (MM) cell adhesion to fibronectin (FN), mediated via VLA-4 and VLA-5, has been shown to induce resistance to several chemotherapeutic drugs. Disruption of MM cell adhesion to FN and other marrow microenvironment elements might therefore enhance the effects of therapy. We now present the first evidence that Eph-ephrin signaling may be exploited to inhibit MM cell binding to fibronectin. Ephs are transmembrane tyrosine kinases and ephrins are their cell-surface ligands. There are two classes of Ephs and ephrins, A and B. Both Ephs and ephrins can transduce repulsive signals that cause interacting cells to lose contact with each other and with extracellular matrix. We are not aware of any previous systematic study of Eph and ephrin expression or function in MM cells. We have found MM cell lines H929, U266, and RPMI 8226 express members of the A classes of both Ephs and ephrins, but not the B classes. First, we demonstrated ligation with commercially available anti-ephrin A3 antibody was followed by ephrin capping and shedding from the cell surface. We next explored whether ephrin ligation affects MM cell adhesiveness in culture. Whereas H929, U266, and RPMI 8226 cells adhered rapidly to fibronectin-coated plastic surfaces, all three cell lines failed completely to adhere to a mixed coating of FN and rabbit anti-ephrin A3 antibody for a period of 2 hrs. This effect was not seen with FN + normal rabbit Ig. This suggests binding of ephrin A3 (or another cross-reacting A-class ephrin) by solid-state antibody triggers intracellular signals that interfere with initial steps of integrin-mediated adhesion. After 2 hr, spontaneous partial recovery of adhesion occurred, reaching a plateau of approximately 30% of control values by 24 hr. We postulate this recovery occurs via clipping of the extracellular ephrin domain by transmembrane metalloproteases, since recovery of FN adhesion was partially prevented by the metalloprotease inhibitor GM6001 (25 uM). Also consistent with this theory, we found in a separate experiment that GM6001 reduced the shedding of cross-linked A-class ephrins from MM cell lines. In summary, we have demonstrated that manipulation of EPH-ephrin signaling can impair MM-cell adhesion to FN, and that this effect is enhanced by simultaneous inhibition of metalloprotease activity. We are currently studying the effect of A-class ephrin ligation on adhesion-mediated drug resistance in MM cell lines. We also intend to evaluate EPH-ephrin expression in marrow specimens from patients with MM.
Increased Peripheral Platelet Destruction and Caspase-3-Independent Programmed Cell Death of Bone Marrow Megakaryocytes in Myelodysplastic Patients.
Authors: Houwerzijl Ewout J. , Blom Nel R. , van der Want Johannes J.L. , Louwes Henk , Esselink Mariet T. , Smit Jan W. , Vellenga Edo , de Wolf Joost Th.M. 
Published: October 2019

Abstract
To investigate underlying mechanisms of thrombocytopenia in myelodysplastic syndrome (MDS), radiolabeled platelet studies were performed in 30 MDS patients with platelets <100 x 109/L. Furthermore, plasma thrombopoietin and glycocalicin-index (a parameter of platelet or megakaryocyte destruction) were determined. Mean platelet life (MPL), corrected for the degree of thrombocytopenia, was reduced in 50% of the patients (4.3 ± 0.9 days (mean ± SD) vs 6.0 ± 1.3, p=0.0003). Platelet production rate (PPR, i.e. the number of platelets entering the circulation to maintain the platelet count) was reduced in 83% (68 ± 34 x 109/d vs 220 ± 65, p<0.0001), although 80% of these patients had normal or increased numbers of bone marrow megakaryocytes. Thrombopoietin levels (patients 223 ± 194 vs controls 118 ± 33 pg/ml, p=0.04) were not significantly correlated with the PPR. However, the glycocalicin-index was significantly higher compared to controls (15 ± 16 vs 0.7 ± 0.2, p=0.001) and significantly and inversely correlated with the PPR (p=0.02, r=−0.5), but not with the MPL (p=1.8). These findings indicate to ineffective thrombopoiesis as a result of increased destruction of platelets and megakaryocytes in the bone marrow leading to a diminished release of platelets to the peripheral blood (i.e. PPR). To investigate whether programmed cell death (PCD) of bone marrow megakaryocytes might be responsible for this ineffective thrombopoiesis ultrastructural studies were performed in 5 patients MDS patients. In the majority of megakaryocytes (66 ± 15%) ultrastructural features of necrosis-like PCD were found, without signs of apoptotic or apoptotic-like PCD, especially in mature megakaryocytes. The data correspond with a negative immunohistochemical staining for activated caspase-3 of bone marrow megakaryocytes. These results indicate that the main cause of thrombocytopenia in MDS might be caspase-3-independent necrosis-like PCD resulting in a decreased PPR in conjunction with an increased glycocalicin-index.
A Phase I/II Multicenter, Safety and Efficacy Study of Combination Treatment with Melphalan, Arsenic Trioxide and Vitamin C (MAC) in Patients with Relapsed or Refractory Multiple Myeloma.
Authors: Berenson James R. 
Published: October 2019

Abstract
Background: An urgent need exists for new treatments to overcome chemoresistance in MM patients. Recent in vitro and In vivo studies in our laboratory show that arsenic trioxide (ATO) can sensitize chemoresistant MM cells to melphalan-induced cytotoxic effects. Pre-clinical studies also show the most profound anti-MM effects when ATO, ascorbic acid and melphalan are used in combination compared with the effects observed when the drugs are used alone or combinations of any two of these agents. Based on encouraging results from a pilot study1, a larger, multicenter trial was recently started.
Methods: MM patients who showed relapse after responding to first-line chemotherapy and/or having proved to be refractory to chemotherapy are enrolled. Patients received a loading dose of ATO at 0.25 mg/kg IV followed by ascorbic acid 1 g IV days 1–4 of week 1 of each six-week cycle. ATO/ascorbic acid was given twice-weekly for the next 4 weeks of each cycle. Low-dose melphalan (0.10 mg/kg) was administered orally for the first 4 days of each cycle. Patients received a maximum of 6 cycles followed by weekly maintenance treatment with weekly ATO followed by ascorbic acid. The primary objectives of this study are to determine response rate and safety and tolerability of MAC therapy.
Results: Twenty patients have received at least one treatment cycle. Preliminary data show that eight (4 PR, 4 MR) of the 14 evaluable patients (57%) had an objective response, an additional three patients achieved stable disease, resulting in a total of 11 patients (79%) with disease control. Since responses were seen after 2 to 5 treatment cycles, it is possible that some patients with stable disease may experience additional disease response. Seven of the eight responding patients had failed two or more treatments: five patients had received prior thalidomide therapy, two had received melphalan and bortezomib, and two patients had undergone autologous peripheral stem cell transplantation. Of the six patients who have now completed the maximal numbers of cycles, four achieved PR, one MR, and one SD. The regimen was well tolerated with few significant side effects reported; mild cytopenias were reported as reversible.
Conclusions: These preliminary results in this treatment group of heavily pre-treated MM patients who had either relapsed or were refractory to standard and/or investigational multiple myeloma treatments suggests that the MAC treatment regimen (1) shows efficacy using a low dose of melphalan supporting the preclinical evidence that ATO can sensitize tumors to chemotherapy; (2) is well tolerated; (3) may require multiple cycles before response.
A Monoclonal Antibody Specific for Free Human Kappa Light Chains Induces Apoptosis of Multiple Myeloma Cells and Exhibits Anti-Tumor Activity In Vivo.
Authors: Asvadi Parisa , Jones Darren R. , Dunn Rosanne D. , Choo Andre B.H. , Raison Matthew J. , Raison Robert L. 
Published: October 2019

Abstract
Despite high dose chemotherapy and autologous stem cell transplant, multiple myeloma (MM) remains an incurable malignancy, with a median 5 year survival of less than 20%. With the exception of idiotype, few antigen targets have been identified that would facilitate specific immunotherapy of MM. We have previously described a murine monoclonal antibody that recognizes a conformation-dependent epitope on free human kappa light chains and a cell surface antigen, KMA, expressed on kappa MM plasma (MMκ) cells (Raison, RL and Boux, HA Mol Immunol 1985 22:1393). Here we show that the murine antibody, designated mKap, bound specifically to a range of MMκ cell lines and inhibited the in vitro growth of these cells. Flow cytometric analysis (Annexin-V and PI staining) of MMκcell lines incubated with mKap demonstrated a dose dependent induction of apoptosis. Furthermore, the presence of activated caspases in mKap treated cells was detected using the CaspACE™ FITC-VAD-FMK reagent. The induction, by mKap, of apoptosis in MMκ cells occured in the absence of cross-linking second antibody or effector cells. In vivo, anti-tumor activity by mKap was demonstrated in a SCID mouse tumor xenograft model. Tumor growth was measured by quantitation of secreted myeloma Ig over a period of 6 weeks. From week 4 onwards, significantly lower serum concentrations of myeloma Ig were detected in animal groups receiving 3.0, 1.5, 0.3 and 0.15 mg total mKap compared to the untreated control (P<0.005 at week 4; P<0.0001 at week 5). The tumor-restricted specificity of mKap, coupled with its ability to inhibit MMκ cell growth in vitro and in vivo, suggests the potential use of either a chimeric or humanised version of this antibody, alone or in combination with chemotherapy, for the treatment of kappa MM.
Genes Regulated by NPM-ALK Fusion Kinase Play a Key Role in the Activation of AP-1 Transcription Factors.
Authors: Staber Philipp B. , Linkesch Werner , Schauer Silvia , Moser Gerit , Kadin Marshall E. , Dirks Willi , Hoefler Gerald 
Published: October 2019

Abstract
Background: About half of nodal anaplastic large cell lymphomas (ALCL) express the nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) fusion protein, which is the product of a t(2;5)(p23;q35) chromosomal translocation. Expression of this protein has been shown to result in neoplastic change. Combining suppression subtractive hybridization (SSH) and cDNA microarray analysis we aimed at elucidating the consequences of NPM-ALK expression.
Methods: SSH cDNA libraries were constructed using mRNA from human embryonic kidney (293) cells transfected with active or kinase-dead NPM-ALK constructs, as well as pools of NPM-ALK positive and negative ALCL cell lines. The resulting cDNA clones and genes relevant for cancer pathogenesis were spotted, generating specific cDNA microarrays comprising 4992 genes. mRNA expression patterns were analyzed in individual cell lines. Real time quantitative RT-PCR of 20 selected genes validated the mRNA expression data of the microarrays.
Results: Expression of a set of 102 genes distinguishes NPM-ALK-negative (FE-PD, MAC-2A) from NPM-ALK-positive ALCL cell lines (SU-DHL-1, JB-6, SUP-M2, SR-786, DEL and Karpas 299). The majority are involved in regulation of cell cycle and apoptosis. 38 of these genes also discriminate 293 cells with respect to their NPM-ALK expression. Interestingly, AP-1 target genes, such as GM-CSFRA, GM-CSFRB, ARF5, FAS, FASL, and BCL3, are increased in NPM-ALK expressing cell lines. Electrophoretic mobility shift assay (EMSA) verifies NPM-ALK dependent AP-1 DNA binding activity.
Conclusion: This study reveals genes specifically regulated by NPM-ALK lymphoma kinase. Further, we demonstrate that AP-1 activation is a critical target of NPM-ALK signalling.