bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Genetic and Expression Markers in DLBCL Cell Lines That Influence the In Vivo Response of DLBCL to Doxorubicin.
Authors: Houldsworth Jane , Petlakh Marina , Chaganti R. S. 
Published: October 2019

Abstract
Diffuse large B-cell lymphoma (DLBCL) is the largest aggressive group of non-Hodgkin’s lymphoma (~40%) and exhibits great clinical and pathological heterogeneity. While DLBCL represents a chemosensitive disease, ultimately, only approximately 50% of patients are cured with current treatment options. Standard front-line therapy comprises CHOP and more recently Rituximab-CHOP. CHOP commonly contains doxorubicin as the curative anthracycline. Controversy currently exists in the use of genetic and expression markers in predicting response to treatment and outcome of DLBCL, thus, we examined the cellular responses of a panel of eight DLBCL cell lines to doxorubicin in order to define molecular markers that influence the response. Expression profiling and subsequent classification of the cell lines revealed that six of the lines were of a germinal center B-cell-like (GCBL) subtype and two (Ly3 and Ly10) were of a non-GCBL subtype. Ly3 and Ly10 exhibited a rapid apoptotic response within 24 hours after exposure to doxorubicin as evaluated by Hoescht staining. Expression profiling of the response of Ly10 cells (in triplicate) revealed the up-regulation of transcripts consistent with mediation of the cellular response via a p53-dependent pathway. Mutation analysis and Western blotting revealed wild type TP53 in Ly3 and Ly10 with an induction of p53 levels after doxorubicin. The remaining cell lines exhibited mutated TP53 sequences and no induction of p53 by doxorubicin. Therefore, in DLBCL cell lines, wild type p53 effects an early rapid apoptotic response to doxorubicin. In the six non-GCBL cell lines, a decrease in viability after doxorubicin was observed at much later times (up to 48 hours after treatment) by both apoptotic (exemplified by Ly2 and Ly8) and non-apoptotic mechanisms (exemplified by Ly1). Expression profiling of these three cell lines revealed alterations in expression of transcripts consistent with a G2/M cell cycle delay induced by doxorubicin, which was confirmed by follow-up flow cytometric analysis. An induction of transcripts consistent with NF-κB and AP-1 activation was observed in Ly1, but not in Ly2 and Ly8, which was confirmed by electrophoretic mobility shift assays. In order to determine if this cellular response plays a role in determining the response of DLBCL to doxorubicin in vivo, the expression profiles of untreated Ly1 and Ly2 were compared, and the resulting 160 differentially expressed transcripts were used to cluster 41 newly-diagnosed DLBCL specimens which had previously been profiled and had a known response to doxorubicin-containing therapy. This analysis revealed a marked correlation between capacity to induce NF-κB and AP-1 with in vivo treatment failure. Overall, these experimental studies have begun to outline genetic and biologic features that together contribute to the response of DLBCL to doxorubicin in vivo.
Efficacy and Safety of Fondaparinux (ARIXTRA®) in the Initial Treatment of Venous Thromboembolism in Obese Patients.
Authors: Buller Harry R. ,   the MATISSE Investigators
Published: October 2019

Abstract
Background: The MATISSE trials demonstrated that once-daily subcutaneous fondaparinux, a synthetic selective factor Xa inhibitor, was at least as effective and as safe as standard therapies in the initial treatment of deep-vein thrombosis (DVT) or pulmonary embolism (PE). Since obesity is a risk factor for venous thromboembolism (VTE), we analyzed the MATISSE data from the subpopulation of obese patients (body-mass index, BMI ≥30 kg/m2).
Methods: Fondaparinux was administered at a once-daily subcutaneous dose of 7.5 mg (5.0 mg and 10.0 mg in patients <50 kg and >100 kg, respectively). In the MATISSE-DVT trial, fondaparinux was compared with twice-daily subcutaneous enoxaparin (1 mg/kg) in patients with DVT. In the MATISSE-PE trial, it was compared with adjusted-dose intravenous unfractionated heparin (UFH) in patients with PE. All drugs were given for at least 5 days and until anticoagulation with oral anticoagulants was therapeutic. The primary efficacy and safety outcomes were recurrent VTE during 3 months’ follow-up and major bleeding (MB) and death during the initial treatment period.
Results: The percentage of obese patients was 26.3% in MATISSE-DVT and 28.7% in MATISSE-PE (Tables). In both MATISSE-DVT and -PE, efficacy and safety data from obese patients were similar to those from non-obese patients (BMI <30kg/m2). The rates of recurrent VTE and MB in obese patients were not significantly different between fondaparinux and either enoxaparin (MATISSE-DVT) or UFH (MATISSE-PE).
Conclusion: Once-daily fondaparinux is at least as effective and as safe as standard therapies in the initial treatment of DVT or PE in obese patients.
MATISSE DVT BMI<30 kg/m2 BMI≥30 kg/m2 Enoxaparin Fondaparinux Enoxaparin Fondaparinux *As treated patients n 791 803 300 280 VTE, n/N 35/791 (4.4%) 31/803 (3.9%) 10/300 (3.3%) 10/280 (3.6%) MB*, n/N 10/788 (1.3%) 12/798 (1.5%) 3/297 (1.0%) 0/279 (0%) MATISSE PE BMI<30 kg/m2 BMI≥30 kg/m2 UFH Fondaparinux UFH Fondaparinux *As Treated Patients n 760 757 322 314 VTE, n/N 35/760 (4.6%) 30/757 (4.0%) 20/322 (6.2%) 12/314 (3.8%) MB*, n/N 8/752 (1.1%) 11/751 (1.5%) 4/314 (1.3%) 2/311 (0.6%)
Rhogtpase RAC2 Is Required for In Vivo Transformation Activity by P210 Bcr-Abl Fusion Oncogene.
Authors: Krishnan Nithya , Bailey Jeff R. , Summey-Harner Victoria , Brunstein Claudio , Verfaillie Catherine M. , Williams David A. 
Published: October 2019

Abstract
Bcr-Abl, the translocation product of the Philadelphia chromosome implicated in human chronic myelogenous leukemia (CML), is a kinase affecting hematopoietic stem cell (HSC) behavior with respect to proliferation, apoptosis, adhesion and migration. Rho GTPases, particularly the Rac subfamily, have been shown to regulate these same cell functions in normal HSC and also regulate gene expression in many mammalian cells. BCR contains a “GTPase-activating protein” domain and a guanine nucleotide exchange domain, the latter or which is preserved in p210 Bcr-Abl. Since HSC functions regulated by Bcr-Abl and Rac are similar, we studied the potential involvement of Rac activation in
Bcr-Abl signaling cascade. Human CML samples demonstrate baseline activation of Rac proteins that is reversed by in vitro treatment with STI571. To study the specific involvement of Rac2, we used a gene targeted mouse model with Rac2 null bone marrow. Using retovirus-mediated gene transfer, we introduced p210 Bcr-Abl in the MSCV vector into wild-type or Rac2−/− HSC/P and studied the behavior of these cells in vitro and in vivo.
Irradiated recipient mice injected with LDBM cells transduced with p210 developed a uniformly fatal myeloproliferative syndrome (Median survival: 45 days, N=12), while mice injected with p210 transduced Rac2−/− LDBM cells (N=12, 2 independent exp.) had 100% survival and no development of leukocytosis, splenomegaly or organ infiltration of hematopoietic cells. These data suggest that Rac GTPases are critical for the transformation of HSC by Bcr-Abl and provide an additional therapeutic target for intervention in CML.
WILD TYPE Rac 2 −/− Empty Vector MSCV-p210 Empty vector MSCV-p210 *p < 0.01 vs WT-MIEG3, **p< 0.01 vs WT-p210 bcr-abl. Proliferation (CPM) Medium 562 ± 278 16,207± 1605* 819.7 ± 363 3,135.5 ± 498** SCF (100ng/ml) 856 ± 187 23,226 ± 2203* 853.7 ± 524 3,756.8 ± 207** Cytokines (SCF, GCSF, MGDF) 8011± 1412 42,711± 13393* 4833 ±1019 3,614.5 ± 1982** Migration (%) Fibronectin 7 ± 0.4 38 ± 1.9* 0.4 ± 0.0 0.8 ± 0.1** SDF-1α 30 ±2.8 13 ±1.1* 0.5 ± 0.0 0.6 ± 0.0** Adhesion (% ) Fibronectin 76± 2.9 40 ±3* 4 ±0.4 10 ±0.1 **
Transplantation with Higher Dose of Natural Killer Cells Associated with Better Outcomes in Terms of Non-Relapse Mortality and Infectious Events after Allogeneic Peripheral Blood Stem Cell Transplantation from HLA-Matched Sibling Donors.
Authors: Sohn Sang Kyun , Kim Dong Hwan , Lee Nan Young , Baek Jin Ho , Kim Jong Gwang , Suh Jang Soo , Lee Kun Soo , Lee Kyu Bo , Shin Ho Jin , Chung Joo Seop 
Published: October 2019

Abstract
Background: Little is known about the role of the CD56+ natural killer (NK) cell dose on the outcome of allogeneic peripheral blood stem cell transplantation (PBSCT). Recently, a higher dose of NK cells has been associated with a lower incidence of severe GVHD in a PBSCT setting. Therefore, the current study attempted to evaluate the effect of the NK cell dose on transplant outcomes, including non-relapse mortality (NRM) and infectious events, in an allogeneic PBSCT setting.
Methods and Materials: Sixty-one cytokine mobilized PBSC recipients from HLA-matched sibling donors were analyzed according to the infused dose of CD34+ cells and NK cells in relation to overall survival (OS), NRM, GVHD, and infectious events.
Results: The group of patients that received a higher dose of NK cells (≥ 5x107/Kg) showed a lower incidence of NRM (p=0.0186) and infectious events (p=0.0107). When confining the analysis to the group that received a CD34+ cell dose of ≥ 6x106/Kg, those patients that received a higher dose of NK cells exhibited a lower incidence of extensive chronic GVHD (p=0.0704). In a multivariate analysis using Cox’s regression model, a higher dose of NK cells was significantly associated with better transplant outcomes (for NRM, NK cell dose p=0.042, for CD34+ cell dose p=0.018; for infectious events, NK cell dose p=0.013, CD34+ cell dose 0.016; for bacterial infection, NK cell dose p=0.049). The group that received a higher NK cell dose also showed a faster immune recovery (p=0.046 for NK cell recovery, p=0.034 for helper T-cell recovery) in serial measurements of peripheral lymphocyte subsets at D+90, +180, and +365.
Conclusions: The present data suggests that a high dose of NK cells may play an important role in improving transplant outcomes, in terms of reducing NRM and infectious events together with CD34+ cells. The protective role of NK cells against infections may also be associated with a faster immune recovery after allogeneic PBSCT.
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The C-Mpl Receptor Expressed on Endothelial Cells Does Not Contribute Significantly to the Regulation of Circulating Thrombopoietin (TPO) Levels.
Authors: Geddis Amy E. , Fox Norma E. , Kaushansky Kenneth 
Published: October 2019

Abstract
Blood levels of TPO, the primary regulator of platelet production, are thought to be regulated primarily by cellular degradation following its binding to, and internalization by its cell surface receptor, c-Mpl. This physiology is supported by in vitro studies, in which TPO can be removed from solution by incubation with platelets, and accounts for the very high levels of circulating TPO in individuals with congenital amegakaryocytic thrombocytopenia (CAMT), as in this disorder there are no c-Mpl receptor-bearing cells to bind and remove the hormone. Previous reports have demonstrated that in addition to hematopoietic cells, c-Mpl is expressed on several types of endothelial cells. We hypothesized that the c-Mpl expressed on endothelial cells would contribute to the regulation of circulating TPO levels. As a model to test this hypothesis, we transplanted 10 Mpl knock-out mice and 10 wild-type (WT) controls with WT marrow cells and analyzed platelet counts and plasma TPO levels over the course of 6 months. The resulting two groups of chimeric mice both express c-Mpl on megakaryocytes and platelets, but only the WT recipients express the receptor on endothelial cells. If c-Mpl receptors normally expressed on endothelial cells take up circulating TPO and degrade it, we expected that the Mpl-null mice reconstituted with WT cells would display increased TPO levels and an increased steady state platelet count compared to the WT recipients. As expected, pre-transplant the platelet counts in the Mpl-null mice were low (average value 398x109/L) and TPO levels were high (average value 8040 pg/ml), as compared to WT controls (platelet counts average 883x109/L, TPO level 745 pg/ml). However, following transplant with WT cells, we found that TPO levels in the chimeric mice were virtually identical (954 pg/ml for the Mpl-null recipients vs. 829 pg/ml for the WT recipients) as were the steady state platelet counts up to 6 months following transplantation (891x109 /L vs. 883x109/Lx109 /L, respectively). These results indicate that the c-Mpl receptor expressed on endothelial cells does not contribute significantly to the regulation of circulating TPO levels or to steady state platelet counts. The apparent lack of TPO adsorbtion by the endothelial cells is surprising, as prior published studies indicate that Mpl receptors expressed on human umbilical vein endothelial cells bind TPO and that c-Mpl is functionally active on hepatic endothelial cells; in addition, the composite surface area of the endothelium is large. These results also imply that patients with CAMT that have successfully engrafted with normal hematopoietic stem cells should have normal (not elevated) TPO levels. Similarly, gene replacement strategies designed to restore c-Mpl in CAMT do not need to be expressed on endothelial cells to establish the normal regulation of TPO.
TLR9 Agonist Immunomodulator Treatment of Cutaneous T-Cell Lymphoma (CTCL) with CPG7909.
Authors: Kim Youn , Girardi Michael , Duvic Madeline , Kuzel Timothy , Rook Alain , Link Brian , Pinter-Brown Lauren , Comerci Carol , McAuley Sonja , Schmalbach Tess 
Published: October 2019

Abstract
CPG 7909 belongs to a new class of chemically defined CpG immunomodulators that target dendritic cell TLR9 receptors inducing IL-12, IFN-gamma, and NK cell function. The rate and durability of response to CPG 7909 was investigated in refractory patients with recurrent or advanced CTCL, who had failed one or more systemic therapies. Dose escalation with weekly sc dosing of patients at 0.08, 0.16, 0.24, or 0.28 mg/kg (3 patients/cohort) for 24 weeks is nearing completion. Additional patients continue to receive treatment at 0.32 (4 patients) or 0.36 mg/kg (12 patients). Clinical response, monitored by Composite Assessment of Index Lesion Disease Severity (CA) and Physician’s Global Assessment of Clinical Condition, has been documented. Of 28 patients enrolled, 7 (25%) have achieved objective clinical response, 5 with partial response (PR) and 2 with complete response (CR). Eleven patients have maintained stable disease (SD), while 10 have had progressive disease (PD). Eight patients have completed 24 weeks of treatment (5 SD, 2 PR, 1 CR) with 12–16 weeks of response while on study. Six patients (3 SD, 2 PR,1 CR) are currently ongoing in the study. Three patients (2 PR, 1 SD) continue to receive long term CPG 7909 at 0.12 mg/kg (58 total doses), 0.28 mg/kg (34 total doses) or 0.32 mg/kg (29 total doses) in a follow on protocol. Responses have been maintained up to 46 weeks.
Weekly doses up to 0.36 mg/kg have been well tolerated. Most reported adverse events have been of CTC grade 1 or 2. The most common are dose-related local injection site reactions (erythema, induration, edema, inflammation and pain) and mild or moderate flu-like symptoms (fatigue, rigors, fever, arthralgia). Four patients had CTC grade 3 drug related AEs: one decreased lymphocyte count (0.08 mg/kg), one increased gamma glutamyl transferase (0.16 mg/kg), one decreased absolute polys (0.36 mg/kg) and one fatigue (0.36 mg/kg).
Enrollment in the phase II portion of the study is ongoing and compares results of patients randomized to receive either 10 mg or 25 mg sc weekly for 24 weeks (equating to effective doses seen in dose escalation).
Clinical Response with CPG 7909 - 16 M, 12 F
Dose N Disease Stage CR PR SD PD 0.36 mg/kg 12 IB (7), IIB, III (3), IVA 0 2 6 4 0.32 mg/kg 4 IIA, IIB, IVA (2) 1 0 1 2 0.28 mg/kg 3 IB (2), III 0 1 2 0 0.24 mg/kg 3 IB, IIB (2) 0 1 1 1 0.16 mg/kg 3 IB (2), IIA 1 1 1 0 0.08 mg/kg 3 IB (2), IVA 0 0 0 3 28 7% 18% 39% 36%
Human Autologous Tumor-Specific T-Cell Responses Induced by Liposome Encapsulated Lymphoma Membrane Proteins.
Authors: Neelapu Sattva S. , Gause Barry L. , Harvey Linda , Frye Andrea R. , Horton Jessie , Robb Richard J. , Popescu Mircea C. , Kwak Larry W. 
Published: October 2019

Abstract
The unique antigenic determinants (Idiotype [Id]) of the immunoglobulin expressed on a given B cell malignancy can serve as a tumor-specific antigen for active immunotherapy. We have previously demonstrated that Id vaccines in follicular lymphoma (FL) patients administered in the minimal residual disease (MRD) state are immunogenic and are associated with induction of complete molecular remissions and long-term disease-free survival (Nature Med 5:1171–1177, 1999). This hybridoma-derived vaccine is now being tested in a pivotal Phase III clinical trial. However, the production of Id protein by hybridoma technology for such vaccine formulation is an expensive and laborious process requiring an average of 3 to 6 months to manufacture the vaccine for each patient. To overcome this difficulty, we developed a novel vaccine formulation where we directly extracted the membrane proteins from lymph node biopsy-derived tumor cells and incorporated them into liposomes along with IL-2. Testing in preclinical studies showed this formulation to be as potent as our prototype hybridoma-derived Id protein vaccine. In the present study, 11 previously untreated and/or relapsed FL patients received 5 injections of this novel vaccine formulation subcutaneously and/or intratumorally at approximately monthly intervals. The vaccine was well tolerated and induced only minor local reactions at the sites of injection. T cell responses were evaluated by cytokine induction and IFNg ELISPOT against autologous tumor. Post-vaccine, but not pre-vaccine, peripheral blood mononuclear cells (PBMC) from 6 out of 10 patients that were assessed, recognized autologous tumor cells, as demonstrated by TNFa, GM-CSF and/or IFNg production. Significant production of cytokines was observed only in response to autologous tumor cells, but not normal B cells. The precursor frequency of tumor-reactive T cells was significantly increased in postvaccine PBMC (range 19–115 IFNg spots/100,000 PBMC), compared with prevaccine PBMC (range 2–7 IFNg spots/100,000 PBMC). Anti-MHC Class I and Class II antibodies inhibited cytokine production suggesting that both CD4+ and CD8+ T cells were involved in the anti-tumor immune responses. Vaccination was associated with induction of a sustained complete response in one patient and correlated with the generation of a potent anti-tumor T cell response. The remaining 10 patients progressed after a median duration of 8 months. We conclude that liposomal delivery of lymphoma membrane proteins is safe, induces tumor-specific CD4+ and CD8+ T cell responses, and may serve as a model for vaccine development against other human cancers. The induction of clinical response warrants further testing of this novel formulation in the setting of MRD where the immunosuppressive effects of the tumor are likely to be least.
Mcl-1 Sequesters Pro-Apoptotic BH3-Only Bim To Inhibit Its Collaboration with Bax in Myeloma Cells.
Authors: Gomez-Bougie Patricia , Clement Monique , Juin Philipe , Bataille Regis , Amiot Martine 
Published: October 2019

Abstract
Multiple Myeloma is a fatal malignancy of B-cell origin characterized by the accumulation of plasma cells within the bone marrow. IL-6 is a major myeloma cell survival factor that induces the activation of both, Ras/MAP kinase and JAK/STAT pathways, the latter being involved in cell survival. Thus, IL-6 starvation or IL-6 signaling blockade by AG490 (a JAK 2 inhibitor) trigger apoptosis. In myeloma cells, others and us have shown that Mcl-1 is the major anti-apoptotic protein, since Mcl-1 antisenses induce apoptosis while Bcl-2 or Bcl-xL antisenses have no effect. In the present study, we examined the pro-apoptotic BH3-only molecule Bim which is implicated in apoptosis induced by growth factor deprivation. The three major Bim isoforms (EL, L and S) are expressed in viable human myeloma cell lines and are negatively regulated by IL-6. Blockade of IL-6 signaling induces the up-regulation of Bim isoforms concomitant to the down-regulation of Mcl-1. Of major interest, in viable myeloma cells, Bim is strongly associated to Mcl-1, to Bcl-2 and to a lesser extent to the dynein light chain, suggesting that Bim is always complexed in viable cells. Bim/Mcl-1 endogenous interaction is disrupted upon apoptosis induction, either by IL-6 starvation or AG 490 treatment. Of note, Bim/Bcl-2 complex is not affected under the same conditions. In parallel to disruption of Bim/Mcl-1 interaction and Bim release, Bax activation was observed. Microinjection of a peptide comprising the minimal BH3 domain from Bim triggered apoptosis in myeloma cells, confirming that free Bim might exert a direct pro-apoptotic effect in these cells. Therefore, in myeloma cells Mcl-1 neutralises Bim through endogenous complex formation. Under apoptosis induction, Mcl-1 drastic down-regulation leads to Bim release, which in turn exerts its pro-apoptotic function through Bax activation.