bluuebunny/crossref_metadata_2025_split huggingface dataset:
Repetitive DNA Hypomethylation in Multiple Myeloma
Authors: Fabris Sonia , Bollati Valentina , Mosca Laura , Pegoraro Valeria , Ronchetti Domenica , Ciceri Gabriella , Baldini Luca , Morabito Fortunato , Deliliers Giorgio Lambertenghi , Bertazzi Pier Alberto , Baccarelli Andrea , Neri Antonino 
Published: April 2020

Abstract
Multiple myeloma (MM) is a malignant proliferation of bone marrow plasma cells characterized by a wide spectrum of genetic and epigenetic changes. Global hypomethylation of repetitive genomic sequences such as long interspersed nuclear elements-1 (LINE-1) and Alu repetitive elements (approximately 500.000 and 1.4 million in the human genome) has been associated with chromosomal instability. Additionally, satellite alfa DNA (SAT-alpha DNA) hypomethylation has been reported to be associated to karyotypic instability in human cancer, possibly playing a role in centromere function. So far, the LINE-1/Alu and centromeric SAT-alpha DNA methylation patterns have not been investigated in the context of the different clinical and molecular MM subtypes. Global DNA methylation changes were investigated in a panel of 53 newly diagnosed, untreated MMs, 7 plasma cell leukemias (PCL) and 11 healthy subjects as controls. DNA was extracted from purified plasma cells, treated with bisulfite and analyzed by bisulfite-PCR and Pyrosequencing. Methylation of LINE-1 and Alu elements was shown to correlate with total 5mC content and thus used to estimate global DNA methylation. MMs showed a decrease of Alu (21.1%) and LINE-1 (70.0%) methylation average levels compared with controls (25.2% and 79.5% respectively). Lower median methylation levels were also found in centromeric SAT-alpha DNA of MMs (77.95%) compared to controls (89.5%). The median methylation level of PCLs was lower than MMs (16.7% versus 21.1% for Alu; 45.5% versus 70.0% for LINE-1; and 33.3% versus 77.9% for SATalpha DNA). Notably, a statistically significant association between SAT-alpha DNA and LINE-1 methylation (Spearman’s rank correlation, ρ = 0.94; P &lt; 0.001) was found in MM. The comparison between methylation pattern and different molecular MM subgroups by means of non parametric tests, revealed that LINE-1 and SAT-alpha DNA methylation was significantly lower in the nonhyperdiploid versus hyperdiploid (HD) tumors (P = 0.01 and 0.02 respectively). Alu and SAT-alpha were significantly lower in the MMs with t(4;14) (P = 0.02 and 0.004 respectively). Finally, in the context of translocation/cyclin D (TC) classification, a statistically significant differences inside the five different groups were found for SAT-alpha DNA methylation (P = 0.008, Kruskal-Wallis test). These findings may provide insights into the molecular mechanisms of MM pathogenesis and suggest that our approach may contribute toward a more exhaustive stratification of the disease.
Identification of New Upregulated Genes with Possible Relevance for Multiple Myeloma Tumorigenesis
Authors: Felix Roberta Spetic , Colleoni Gisele W. B. , Caballero Otavia L. , Almeida Manuella Sampaio , Andrade Valeria C.C. , Chauffaille Maria L. L. F. , Silva Wilson A. , Begnami Maria Dirlei , Soares Fernando Augusto , Zago Marco Antonio , Vettore Andre Luiz 
Published: April 2020

Abstract
Serial analysis of gene expression (SAGE) allows a comprehensive profiling of gene expression within a given tissue and also an assessment of transcript abundance.
Objectives: We generated SAGE libraries from normal and neoplastic plasma cells to identify genes differentially expressed in multiple myeloma (MM).
Material and Methods: Normal plasma cells were obtained from palatine tonsils and MM SAGE library was generated from bone marrow plasma cells of MM patients.
Results: We obtained 29,918 SAGE tags from normal and 10,340 tags from tumor libraries. Computer-generated genomic analysis identified 46 upregulated genes in the MM library. Ten upregulated genes were selected for further investigation. Differential expression was validated by quantitative real-time PCR in purified plasma cells of 31 patients and three controls. P53CSV, DDX5, MAPKAPK2, RANBP2 were found to be upregulated in at least 50% of the MM cases tested. All of them were also found upregulated in MM when compared to normal plasma cells in a meta-analysis using ONCOMINE microarray database. Antibodies specific to DDX5, RANBP2 and MAPKAPK2 were used in a TMA containing 57 MM cases and confirmed the expression of these proteins in 74, 96, and 21% of the MM samples, respectively.
Conclusions: Analysis of differential expression using SAGE could identify new genes important for myeloma tumorigenesis (P53CSV, DDX5, MAPKPK2 and RANBP2) and that could potentially be useful as therapeutic targets.
CCR1 Inhibition Targets Osteoclast-Myeloma Adhesion and Exerts in Vivo Anti-Osteoclast Activity
Authors: Vallet Soni a , Vaghela Nileshwari , Fulciniti Mariateresa , Veiby Petter , Hideshima Teru , Pozzi Samantha , Santo Loredana , Mukherjee Siddhartha , Cirstea Diana , Scadden David T. , Anderson Kenneth C. , Raje Noopur 
Published: April 2020

Abstract
Among the components of the tumor microenvironment, osteoclasts (OC) and bone marrow stromal cells (BMSC) play a key role in stimulating multiple myeloma (MM) cell proliferation via cell-to-cell contact and cytokine secretion. In a vicious loop, MM cells promote OC development and impair osteoblasts (OB) differentiation shaping the microenvironment to support tumor growth and chemoresistance. These effects translate in osteolytic lesions that negatively affect patient quality of life and survival. CCL3 is involved in the pathogenesis of MM bone disease. Indeed, high bone marrow plasma levels of CCL3 correlate with osteolytic lesions in MM and CCL3 has a pro-osteoclastogenic effect. We have previously shown that inhibition of CCL3 receptor, CCR1, results in impaired osteoclastogenesis and blocks the proliferative advantage conferred by OCs to MM cells in vitro. (Vallet et al, Blood 2007) Here, we extend our studies and demonstrate in an in-vivo SCID-hu mouse model the anti-OC effects of MLN3897, a small molecule CCR1 inhibitor (Millennium Pharmaceuticals, Cambridge). CB17 SCID mice bearing a human fetal bone implant engrafted with INA6 MM cells were treated orally with MLN3897 for a total of 49 doses. After 4 weeks of treatment the bones were harvested and stained for TRAP activity and hematoxylin-eosin. The number of OC/400x field was significantly reduced in the treated group (2.7 vs 1.9, p&lt;0.05), thus confirming in vivo the anti-osteoclastogenic effect of CCR1 inhibition. Although a 2-fold inhibition of tumor growth in the treated group was noted, this difference did not reach statistical significance. Our previous data suggested that CCR1 inhibition impaired OC-MM cell adhesion with subsequent reduced MM cell proliferation. Here, we observed a specific effect of CCR1 inhibition on MM cell/OC adhesion rather than BMSCs and fibronectin. Interestingly, these effects translated in anti-proliferative effects of the CCR1 inhibitor in OC-MM cells coculture with no effects in BMSCs-MM coculture, suggesting a specific inhibitory effect on the OC compartment. Ongoing studies to characterize the underlying mechanism of CCR1 inhbition on OC-MM cell adhesion will be presented. Combination approaches to study whether forced OB differentiation by the proteasome inhibitor, bortezomib, will improve BMSC sensitivity to CCR1 inhibition are being studied. These data confirm the specific in vivo anti-OC effects of CCR1 inhibition and will highlight the role of novel combinations by using anti-OC agents like MLN3897 and anabolic drugs like Bortezomib.
Diffuse Large B-Cell Lymphoma (DLBCL) Patients with Composite Histology Have Improved Early Outcome Compared to De Novo DLBCL When Treated with R-CHOP
Authors: Maurer Matthew J. , Witzig Thomas E. , Macon William R. , Syrbu Sergei I. , Cerhan James R. , Allmer Cristine , Johnston Patrick B. , Link Brian K. , Habermann Thomas M. 
Published: April 2020

Abstract
Background: A significant percentage of DLBCL patients present with a composite histology, often seen as a node containing both follicular lymphoma and DLBCL or DLBCL in the node and discordant indolent lymphoma in the bone marrow. Literature from the pre-rituximab era suggests DLBCL patients with transformed lymphoma or composite histology have worse outcome than de novo DLBCL. Here we report on early events in a cohort of R-CHOP treated patients.
Goal: To determine whether patients with composite lymphoma have an inferior event free survival (EFS) and overall survival (OS) compared to de novo diffuse large B-cell lymphoma when treated with R-CHOP.
Methods: Newly diagnosed patients treated with an R-CHOP containing regimen were prospectively enrolled in our Lymphoma SPORE registry from 9/2002 through 6/2007. Pathology was centrally reviewed. All patients were followed for retreatment, disease progression, and death.
Results: 401 DLBCL patients were enrolled; 14% (57/401) had a composite histology. 33 patients had DLBCL and another histology, predominantly follicular lymphoma (n=29), in the same node. 20 patients had a non-DLBCL histology in a distinct location from the DLBCL; this was primarily indolent lymphoma in the bone marrow (n=15). 4 patients had both. 19% (75/401) of patients died during follow-up and 30% (121/401) had an event (death due to any cause, progression, or retreatment). Median follow-up for living patients was 34 months (range, 5–73). Composite DLBCL patients had higher event-free (3 year EFS = 79%) and overall (3 year OS = 93%) survival then de novo DLBCL (3 year OS = 66%, 3 year EFS 79%), p=0.05 and p=0.005 respectively. These differences remained statistically significant after adjusting for the International Prognostic Index (IPI): EFS HR = 0.53, 95% CI: 0.29–0.97, p=0.02; OS HR=0.28, 95% CI: 0.10–0.76, p=0.002. OS and EFS for composite DLBCL more closely resembled R-CHOP treated grade III follicular lymphoma (A,B) from the same cohort (3 year EFS = 81%, 3 year OS = 93%). Improved outcome for composite DLBCL was consistent whether the additional histology was in the same node or distinct from the DLBCL.
Conclusions: R-CHOP treated DLBCL patients with indolent discordant bone marrow involvement or other composite histology have improved early OS and EFS compared to de novo DLBCL. Further follow-up is needed to assess the long-term prognosis of composite DLBCL in the rituximab era.
Histology N Age &gt; 60 Stage III/IV LDH &gt; ULN PS &gt; 1 &gt;2 EN Sites 3 YR EFS 3 YR OS * Denotes statistically significant difference at p=0.05 de novo DLBCL 344 58% 56% 56% 17% 22% 66% 78% Composite DLBCL 57 65% 77%* 34%* 18% 32% 79%* 93%* Figure Figure
T-Cell Receptor Signaling Proteins, BCL10 and Phosphorylated-P65 NFKB, Are Expressed in Peripheral T-Cell Lymphomas and Associated with Clinical Outcome
Authors: Briones Javier , Espinosa Iñigo , Delgado Julio , Sierra J. 
Published: April 2020

Abstract
Background: Peripheral T-cell lymphomas (PTCL) are characterized by T-cell receptor (TCR) signaling alterations and NF-kB activation. Protein kinase C (PKC) is a family of proteins involved in signal transduction and mediates activation of the NF-kB pathway. In normal T cells, PKC 𝛉 plays a major role in TCR-mediated activation of a novel NF-kB pathway that involves phosphorylation of p65 at Serine 536 (P-p65Ser536). BCL10 acts along the same pathway downstream of PKC 𝛉 to activate NF-kB. We thus investigated the relationship between PKC 𝛉, BCL10 and P-p65Ser536 status in biopsy specimens from patients with PTCL.
Methods: Paraffin-embedded tissues obtained at diagnosis from 30 patients with nodal PTCL (PTCL unspecified, 21cases; angioimmunoblastic, 3; ALK− anaplastic large-cell, 6) treated with curative intention were evaluated. Expression of PKC 𝛉, BCL10, and P-p65Ser536 proteins were assessed using immunohistochemistry on a tissue microarray.
Results: Expression of PKC 𝛉 was detected in 22 of 30 cases (73%), BCL10 in 20 of 30 (67%), and P-p65Ser536 in 21 of 30 (70%). Expression of BCL10 was associated with PKC 𝛉 (18 of 22) (P&lt;0.0001), and P-p65Ser536 (19 of 21) expression (P&lt;0.0001). After a median follow-up of 60 months for surviving patients (range, 21–160 months), 5-year overall survival (OS) was 33%. Patients with BCL10 or P-p65Ser536 positive tumors fared better, with a 5-year OS of 48% and 45%, respectively, versus 0% for those with negative tumors (P=0.029, and P=0.04, respectively). Remarkably, BCL10 expression identified a subgroup of patients within the low Ki-67 group (&lt;80% positive tumor cells) with a different outcome; thus, within this subgroup patients with BCL10 positive tumors had a 5-year OS of 85% while it was 0% for those with BCL10 negative tumors (P=0.011). Multivariate analysis showed that BCL10, Ki-67, and P-p65Ser536 expression were independent factors associated with OS (P=0.039, 0.029, and 0.050, respectively).
Conclusion: BCL10 is expressed in PTCL, correlates with PKC 𝛉 and P-p65Ser536 NF-kB expression and it seems to be associated with improved survival in PTCL.
Clearance Mechanisms for Chilled Platelets
Authors: Rumjantseva Viktoria , Josefsson Emma C , Grewal Prabhjit K , Wandall Hans H. , Neyeb-Hashemi Sara , Larson Goran , Marth Jamey D , Hartwig John H. , Hoffmeister Karin M. 
Published: April 2020

Abstract
When platelets are chilled for short times (&lt;4 hours), βN-acetylglucosamine (βGlcNAc) residues of N-linked glycans on clustered GPIbαmolecules are recognized by the lectin domain of αMβ2 receptors on hepatic macrophages, and are removed from circulation. Clearance can be prevented by galactosylation of the exposed βGlcNAc residues. Galactosylation, however, does not improve the circulation of platelets refrigerated for longer times (&gt;48 h-refrigerated) (Wandall et al. Blood, 2008). Investigating the clearance mechanism of 48 h-refrigerated platelets, we now demonstrate macrophage-independent hepatic removal, that is mediated by the Ashwell-Morell asialoglycoprotein receptor (AMR). This conclusion is based on the following evidence:
macrophage depletion in mice using clodronate encapsulated liposomes, a procedure that dramatically improves the circulation of 4 h chilled platelets, does not prevent the clearance of 48 h chilled platelets; streptavidin-POD staining reveals abundant long-term refrigerated and biotinylated platelets in hepatocytes; 48 h-refrigeration increases by ~1.7-fold the binding of the βgalactose-recognizing lectin RCA, but not of the βGlcNAc recognizing lectin sWGA lectin to platelets; KO mice lacking Asgr-1 or Asgr-2 subunits of the AMR support 48 h-refrigerated platelet circulation times comparable to room temperature stored platelets; Co-injection of asialofetuin, a competitive inhibitor of the asialoglycoprotein-receptors, restore the survival of 48 h-refrigerated platelets in WT mice; and hepatocyte HepG2 cells ingest fluorescently labeled long-term refrigerated platelets in culture, and asialofetuin prevents this ingestion.
Regarding the platelet target of the Asgr-1/Asgr-2 receptors, circulation of 48 h-refrigerated platelets is markedly improved by removal of GPIbα’s N-T domain using O-sialopeptidase. We conclude that longterm refrigeration exposes galactose residues on GPIbα subunits of platelets to generate ligands recognized by hepatocyte Ashwell-Morell receptors.
Engagement of Fibrinogen and β3 Integrin Is Required for Maintenance of Platelet Intracellular and Cell Surface P-Selectin Expression
Authors: Yang Hong , Lang Sean , Zhai Zhimin , Spring Christopher M. , Reheman Adili , Chen Pingguo , Flick Matthew J. , Degen Jay L. , Li Ling , Kahr Walter H.A. , Freedman John , Ni Heyu 
Published: April 2020

Abstract
Platelet P-selectin plays important roles in inflammation and contributes to thrombosis and hemostasis. While it has been reported that von Willebrand factor (VWF) affects P-selectin expression on endothelial cells, little information is available regarding regulation of platelet P-selectin expression. Here, we first observed that P-selectin expression was significantly decreased on platelets of fibrinogen and VWF double deficient mice. Subsequently, we identified this was due to fibrinogen, but not VWF, deficiency. The impaired P-selectin expression on fibrinogen-deficient platelets was further confirmed in a human patient with severe hypofibrinogenemia and his heterozygous parents. We demonstrated that this impairment is unlikely due to excessive P-selectin shedding, deficient fibrinogen-mediated cell surface P-selectin binding, or impaired platelet granule release, but rather is due to decreased platelet P-selectin content. This effect seems to be specific for P-selectin, as other alpha-granule proteins (e.g. thrombospondin-1, vitronectin, platelet factor 4) were not decreased in fibrinogen-deficient mouse or human platelets. We found that fibrinogen transfusion recovered the P-selectin expression in fibrinogen-deficient mice to &gt;80% of the normal level within 48 hours, and four days post-transfusion, the P-selectin levels were nearly completely recovered. Furthermore, engagement of the C-terminus of the fibrinogen γ chain with β3 integrin was required for this process since a similar impairment of P-selectin expression was also observed in fibrinogen γΔ5 mice and β3 integrin-deficient mice. To determine whether fibrinogen affects the biogenesis of platelet alpha-granules, platelets were examined via electron microscopy. No statistically significant difference in the number of platelet alpha-granules was observed in fibrinogen-deficient or β3 integrin-deficient mice compared to controls (P&gt;0.05). These data suggest fibrinogen may play important roles in inflammation, including immune-mediated inflammation, thrombosis and hemostasis via enhancement of platelet P-selectin expression. Furthermore, there are 2–3 fold variations in human plasma fibrinogen levels in healthy populations, and there are patients with hypo- and afibrinogenemia, hyperfibrinogenemia, and Glanzmann’s Thrombasthenia (β3 integrin deficiency). Our data suggest that platelet P-selectin expression can be affected by all these conditions. Therefore, P-selectin as a platelet activation marker should be used with caution.
Oct4 Expression and Mesenchymal Stromal Cell Plasticity
Authors: Yannarelli Gustavo , Jiang Huijie , Montanari Sonia , Helke Simone , Wang Xinghua , Keating Armand 
Published: April 2020

Abstract
Mesenchymal stromal cells (MSCs) have the capacity to differentiate along multiple lineages and are now in cell therapy clinical trials, especially for injured myocardium. Mechanisms mediating tissue regeneration remain unclear and are likely multifactorial. We recently showed that bone marrow-derived murine MSCs can acquire cardiac markers but retain MSCs properties when co-cultured with rat embryonic cardiomyocytes (RECs) (Rose et. al., Stem Cells. Aug7, 2008). The aim of our study was to determine whether expression of the embryonic transcription factor, Oct4 was modulated in this model of MSC plasticity. Wild-type Fvb mouse MSCs (passage 4) were co-cultured with RECs for 5 days and expression of Oct4, Nanog and Sox2, was analyzed by qRT-PCR with mouse-specific primers. Oct4 protein was assessed by immunocytochemistry (ICC) and flow cytometry (FC). The MSCs expressed Oct4, Nanog, and Sox2 transcripts. After co-culture, mRNA levels for Oct4 and Sox2 were upregulated 2.6±1.2-fold (p&lt;0.05) and 2.4±0.4-fold (p&lt;0.05), respectively, compared with MSCs controls, in contrast to Nanog gene expression which remained unchanged (1.2±0.3-fold; p=ns). 83±9% of MSCs nuclei were positive for Oct4 by ICC. To distinguish mouse from rat cells in co-culture, cells were stained with an anti-mouse CD44 antibody which does not cross react with rat CD44. CD44 is expressed on all MSCs and absent on RECs. Flow cytometry showed that Oct4 was over-expressed in CD44+ cells after co-culture. Our data demonstrate that these embryonic transcription factors are constitutively expressed in murine MSCs and that Oct4 and Sox2 transcript levels are increased after co-culture with RECs. Oct4, Nanog and Sox2 are known to maintain pluripotency of embryonic stem cells. Moreover, induced pluripotent stem cells can be generated from mouse fibroblasts by the introduction of Oct4 and Sox2 without the need for Nanog (Cell126:663,2006). Our data infer that embryonic transcription factors are involved in MSCs progression to different cell lineages. Our findings suggest new mechanisms that may mediate MSC plasticity.
IFN-Alpha and gm-CSF Can Reverse Immunomodulatory Effects of Imatinib
Authors: Bund Dagmar , Buhmann Raymund , Kolb Hans Jochem 
Published: April 2020

Abstract
Presently, Imatinib (IM) is the first line therapy for chronic myeloid leukaemia (CML), but in most patients CML recurs after discontinuation of IM. In contrast, allogeneic stem cell transplantation (ASCT) is considered to be the only curative treatment for this malignancy curing by the immune effects of donor T-cells against CML progenitor cells. Recent reports have described that IM has inhibitory effects on both the function of T-cells and antigen presenting cells (APCs) like dendritic cells. In the present study we further characterized the immunomodulatory effects of IM on T-cells and APCs and analysed whether IFN-alpha (interferon alpha, IFN-a) alone or in combination with GM-CSF (granulocyte-macrophage colony-stimulating factor) can influence the negative IM effects in vitro. Patient derived CML cells were examined to stimulate allogeneic HLA-mismatched T-cells without and with Imatinib (1, 2 or 5 micro M). The activation profile (CD25) and the expression of the TCR-alpha (T-cell receptor) on the T-cells was determined by FACS after 5 days in presence of IM as well as the proliferative T-cell response which was evaluated using CFSE (Carboxy Fluoroscein Succinimidyl Ester). In [51Cr]-release assays the cytotoxicity of CD8+ T-cells was assessed. The cytokine profile of the culture supernatants was detected by CBA (Cytokine bead array, Beckton Dickinson, USA). We also characterized the antigen profile (HLA-DR, CD40, CD54, CD58, CD80 and CD86) of the CML cells as APCs over a 5 day culture with and without IM. The rescue of the CML cells was initiated by IFN-a or IFN-a/GM-CSF after 3 days incubation with IM. On day 6 the APC profile of the CML cells was obtained and cells were used as stimulators in terms of T-cell proliferation which was determined by CFSE. We could show that the proliferation of allogeneic T-cell was inhibited in the presence of IM in a dose-dependent manner. In addition, the T-cell activation marker CD25 and the TCR-alpha expression were significantly downregulated by different concentrations of IM. Moreover, IM impaired the cytotoxic function of allogeneic T-cells in a dose-dependent manner. Interestingly, after stimulation with CML-cells cytotoxic T-cell activity could only be induced in those cases that did not secrete IL-6. Down-regulation of the APC profile on CML-cells (adhesion/costimulatory molecules) by increasing concentrations of IM could be in part reversed by the addition of IFN-a and completely restored by the combination of IFN-a and GM-CSF. Although, the proportion of proliferating T-cells could not be increased further by the combination of IFN-a ± GM-CSF, the number of T-cells was increased in the presence of IFN-a + GM-CSF. In conclusion, the down-modulating effects of IM could be almost completely reversed by the addition of IFN-a and GM-CSF. It remains to be seen whether these findings can be successfully applied to the treatment of CML-patients.