bluuebunny/crossref_metadata_2025_split huggingface dataset:
PDE4 Inhibitors Augment Glucocorticoid Receptor Levels in B-CLL Cells but Not in T Cells, B Cells, Monocytes or Neutrophils.
Authors: Lerner Adam , Meyers John A. , Taverna Josephine A. , Makkinje Anthony 
Published: October 2019

Abstract
cAMP analogues have long been known to induce apoptosis in specific lymphoid subsets and to augment the apoptotic effect of glucocorticoids. However, a clinically feasible means by which to take advantage of these effects for the treatment of lymphoid malignancies has not previously been identified. Inhibition of type 4 cAMP phosphodiesterases (PDE4) activates cAMP-mediated signaling in many cell types and several PDE4 inhibitors are currently in clinical trials for inflammatory illnesses such as asthma, COPD and psoriasis. We have previously reported that PDE4 inhibitors induce apoptosis in B-CLL and augment the apoptotic effects of glucocorticoids such as dexamethasone. However, the mechanism by which PDE4 inhibitors increase glucocorticoid-induced apoptosis has remained unclear. In gene chip studies, we find that the PDE4 inhibitor rolipram (10 uM) augments transcript levels of glucocorticoid receptor alpha (GRa). A rolipram-induced dose and time-dependent increase in GRa levels was confirmed both by real-time PCR and by Western analysis. In leukemic cells derived from eight B-CLL patients, GRa transcript levels rose over the first four hours to a mean of 4.8 +/− 0.2-fold above baseline and maintained such a fourfold increase for at least 24 hours. An analysis of GRa transcript half-life following actinomycin D treatment demonstrates that rolipram augments transcript levels in B-CLL cells by a transcriptional mechanism. Consistent with this, rolipram treatment augments GRa transcripts derived from promoters 1A and 1C to a greater degree than 1B. Remarkably, rolipram does not augment GRa transcript levels in normal T cells, B cells, monocytes or neutrophils, even in the presence of the adenylate cyclase activator forskolin. Similarly, no increase in GRa transcript levels was observed in leukemic T-CLL cells. As previously reported in B cell lines, treatment of B-CLL cells with dexamethasone reduced GRa transcript levels in a dose-dependent manner. Co-treatment with PDE4 inhibitors maintained GRa transcript levels above basal levels even at the highest dexamethasone concentration tested (1 uM). Finally, we have confirmed the ability of PDE4 inhibitors to augment dexamethasone-induced apoptosis using several PDE4 inhibitors currently in clinical trials for other indications. Our results suggest that co-treatment with PDE4 inhibitors and high-dose dexamethasone may increase leukemic cell GRa levels and augment the apoptotic efficacy of glucocorticoid therapy in B-CLL patients.
Clinical Features and Prognosis of Patients with Myelodysplastic/Myeloproliferative Syndrome-Unclassified (MDS/MPD-U): Refractory Anemia with Ringed Sideroblasts with Thrombocytosis (RARS-T) Is a Favorable Prognostic Subgroup.
Authors: Atallah Ehab , Kantarjian Hagop , Cortes Jorge , Pierce Sherry , Estey Elihu , Garcia-Manero Guillermo , Thomas Deborah , Ravandi-Kashani Farhad , Giles Francis , Verstovsek Srdan 
Published: October 2019

Abstract
MDS/MPD-U is a newly defined clinical entity in the WHO classification. It includes patients (pts) with features of both myelodysplasia and myeloproliferation that cannot be classified under any other WHO category. In addition it includes the provisional entity RARS-T. Recent reports found an increased incidence of JAK-2 mutation in pts with RARS-T, suggesting possibly a different biological behavior. The clinical outcome of MDS/MPD-U, and in particular RARS-T, pts have not been well described.
We reviewed 142 patients in the MD Anderson Leukemia Department patient database with the diagnosis of MDS/MPD or MPD-U seen from September 1987 to July 2006, and identified 32 pts that fulfilled MDS/MPD-U criteria according to WHO classification. The median age was 64 years (range 21–82), with a male: female ratio of 3:1. Six pts had RARS-T (19%). Six pts had a previous malignancy (19%), of which 3 received radiation therapy; no chemotherapy was given. Twenty-five pts (73%) were symptomatic at presentation, including fatigue in 17 (53%), weight loss in 13 (41%), fever in 2 (6%), and night sweats in 7 (22%) pts. Fifteen (45%) pts were transfusion dependent. The median hemoglobin was 9.6 g/dl (range 4.8–15.3), white blood cell count 9x109/L (range 0.6–141), platelets 234x109/L (range 7–1687), and percentage of bone marrow blasts 3% (range 0–16). Fifteen pts (45%) had more than one lineage dysplasia and 22 pts (69%) had increased reticulin in the bone marrow. Cytogenetic analysis revealed diploid cytogenetics in 13, trisomy 8 (either alone or with other abnormalities) in 8, complex cytogenetics in 3, and others in 8 pts. Twenty-two pts were treated with multiple different therapies, including hypomethylating agent in 8, with no sustained responses to any therapy. Six pts received supportive care only and 4 are unknown. Two pts underwent bone marrow transplant; one is in remission for >4 years and the other died from acute graft versus host disease. Median follow up time for alive pts was 10.6 months (range 0.3–147). Median survival of the whole group was 19 months (95% CI; range 8–29). Of the 6 pts with RARS-T, 5 had diploid cytogenetics, two had prior malignancy and 5 were transfusion dependent at the time of diagnosis. Patients with RARS-T had a significantly better survival than the other pts (median survival not reached vs. 12 months; p=0.019; Figure 1).
In conclusion, pts with MDS/MPD-U have a poor prognosis with a median survival of one year. However, RARS-T represents a subgroup of pts with better prognosis and long overall survival justifying their separate categorization.
Overall Survival Overall Survival
Both the Endoplasmic Reticulum and the Mitochondria Are Involved in Apoptosis of Erythroid Precursors in Low Grade Myelodysplastic Syndromes.
Authors: Gyan Emmanuel , Frisan Emilie , Deschemin Jean-Christophe , Randriamamptita Clotilde , Dubart-Kupperschmitt Anne , Beyne-Rauzy Odile , Dreyfus Francois , Guesnu Martine , Lacombe Catherine , Mayeux Patrick , Solary Eric , Fontenay Michaela 
Published: October 2019

Abstract
Cell death by apoptosis accounts for the ineffective erythropoiesis that characterizes low grade myelodysplastic syndromes (LG-MDS). We have shown that the death receptor Fas was over-expressed at the surface of LG-MDS erythroid precursors. In an ex vivo culture of bone-marrow CD34+ cells undergoing differentiation into red cells, we observed that apoptosis of LG-MDS erythroid precursors resulted from Fas-mediated activation of caspase-8. Both a Fas-Fc construct and the ectopic expression of a dominant negative mutant of the adaptor FADD (Fas-Associated Death Domain) could prevent the death of MDS erythroid precursors.
In this ex vivo culture system, apoptosis of erythroid precursors was associated with the release of cytochrome c from the mitochondria to the cytosol. To further address the mitochondria involvement in this apoptotic process, we ectopically expressed, by the use of a lentiviral vector, the anti-apoptotic protein Bcl-2 in MDS (n=20) and normal (n=10) CD34+ cells before inducing their erythroid differentiation. Bcl-2 expression delayed the erythroid differentiation of MDS as well as normal CD34+ cells. It also prevented phosphatidylserine exposure, mitochondrial membrane permeabilization, cytochrome c release, caspase-9 and -3 activation in LG-MDS erythroid precursors. Together with the cleavage of Bid, specifically observed in LG-MDS samples, these observations indicated that the Fas-mediated pathway that led to apoptosis of LG-MDS erythroid precursors was connected to the mitochondrial pathway to death.
We also addressed the role of the endoplasmic reticulum (ER) in this death pathway by using a lentiviral vector encoding an ER-targeted Bcl-2 construct, flag-Bcl2-Cb5. The corresponding protein co-localized with the ER protein calreticulin. ER-targeted Bcl-2 prevented apoptosis induced by thapsigargin, an inhibitor of endoplasmic/sarcoplasmic ATPases (SERCAs), without inhibiting apoptosis induced by lonidamine, a mitochondria targeting agent. Like wild-type Bcl-2, ER-targeted Bcl-2 inhibited phosphatidylserine exposure, mitochondrial membrane permeabilization, cytochrome c release, caspase-9 and caspase-3 activation that characterize spontaneous apoptosis of LG-MDS erythroid precursors in culture. Although ER calcium stores were increased in LG-MDS erythroid precursors, the release of calcium was not modulated by ER-targeted Bcl-2 in these cells. Altogether, our results indicate that, in LG-MDS erythroid precursors, spontaneous apoptosis starts at level of the death receptor Fas, then connects to the mitochondrial pathway that functions downstream of the ER through a calcium-independent and Bcl-2-regulated pathway.
Abrogation of the DNA-Damage Response in De Novo AML Versus MDS Cell Lines.
Authors: Boehrer Simone , Ades Lionel , Fabre Claire , Fenaux Pierre , Kroemer Guido 
Published: October 2019

Abstract
Background: The increasing evidence that activation of the DNA-damage-response (DDR) pathway in preneoplastic lesions of solid neoplasms hinders tumorigenesis prompted us to evaluate the same hypothesis in cell line models representing different subtypes of MDS or AML.
Methods: myeloid cell lines corresponding to different subgroups of MDS, having or not progressed to AML (p39, MOLM-13) and de novo AML (MV4-11, and KG-1) were assessed for their capacity to induce a DDR. Using the quintessential inducer of a DDR, the cell lines were gamma-irradiated (2, 5 and 10Gy) and their ability to arrest the cell cycle in G2/M and to undergo apoptosis were assessed.
Results: in contrast to the de novo AML-derived KG-1 cells - the irradiated MDS/AML-derived cell lines P39 and MOLM-13 exhibited an earlier and more pronounced arrest of the cell cycle in G2/M, as well as a higher percentage of apoptotic cells. Whereas 10 Gy induced a G2/M-arrest in about 30% of KG-1 cells, this percentage increased to about 60% in P39 and MOLM-13 cells. Comparably, whereas KG-1 cells exhibited about 15% of apoptotic cells following irradiation with 10Gy, the percentage reached 30% in P39 and MOLM-13 cells. Assessing the underlying molecular determinants we demonstrated that irradiated P39 and MOLM-13 cells retained the ability to activate crucial mediators of the DDR, whereas this ability was decreased in the de novo AML cell line KG-1. In particular, in comparison to KG-1 cells, irradiated P39 and MOLM-13 cells exhibited a more pronounced upregulation of the crucial cell cycle/checkpoint regulators Chk1-P-Ser317 and Cdk1-P-Tyr15. Noteworthy, MDS cell lines with an increased G2/M-arrest exhibited a higher expression of nucleophosmin - which serves as a shuttling protein between the nucleus and the cytoplasm - and were able to retain this increased expression upon irradiation. Moreover, elevated expression of NPM in MDS cell lines corresponded to higher PCNA-expression, which serves as an essential component of the DNA-repair machinery. To further investigate the functionality of the DDR-pathway, the different cell lines were gamma-irradiated in the presence of the ATM-inhibitor KU-55933 and the Chk1 inhibitor UCN-01. Whereas inhibition of Chk1 (and to a lesser extent inhibition of ATM) was able to hinder G2/M-progression in irradiated MDS cells, it remained without effect in the AML cell lines KG-1 and MV4-11. Noteworthy, whereas irradiated P39/MDS cells hindered to activate Chk1 decrease expression of Polo-like-kinase-1 (Plk-1) - which not only governs mitotic entry and exit, but also functions as an oncogene - KG-1 cells maintained a high expression of Plk-1, comparable to that observed in non-irradiated cells.
Conclusion: we demonstrated the different capacities of MDS (or AML post MDS) and de novo AML cell lines to activate the DDR and thus provide evidence that the DDR-pathway is abrogated in de novo AML as compared to MDS cells.
Clofarabine Is Active in Myelodysplastic Syndrome (MDS).
Authors: Faderl Stefan , Gandhi Varsha , O’Brien Susan , Giles Francis , Cortes Jorge , Plunkett William , Kantarjian Hagop M. 
Published: October 2019

Background. Clofarabine, an adenosine nucleoside analog, is active in acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL). Its anti-MDS activity is not well defined.
Study Aims. To evaluate the safety and efficacy of clofarabine in MDS in two studies, one with intravenous administration and one with oral, using lower doses than those for leukemia (40–52 mg/m2 IV daily x 5).
Study Group and Results. The first study randomized patients in a Bayesian design to receive clofarabine 15 or 30 mg/m2 IV daily x 5 every 4–6 weeks. The second study used clofarabine 40 mg/m2 orally daily x 5 every 4–6 weeks (assuming oral bioavailability about 50%). Standard eligibility criteria for MDS were used; only high risk MDS patients (IPSS intermediate or high) were eligible. So far, 16 patients have been treated, 10 on IV clofarabine and 6 on oral clofarabine. Age ≥60 yr in 13 (81%); prior therapy with decitabine/azacitidine 11 (69%); cytogenetic abnormalities 12 (75%). Responses were graded according to International Working Group criteria (IWG). Results are shown in the table below.
Myelosuppression was significant resulting in febrile episodes and hospitalization in 4/6 patients on oral clofarabine and in 4/10 patients on IV clofarabine. Rash grade 3 was noted in 1 patient on oral clofarabine.
Summary. Clofarabine is active in MDS. The oral dose of 40 mg/m2 daily x 5 may be too high for MDS; lower dose of oral clofarabine 30 mg/m2 orally daily x 5 will be tested with PK studies. The 2 IV schedules (15 vs. 30 mg/m2) are ongoing.
Oral Clofarabine IV Clofarabine • Treated/evaluable to date 6/3 10/6 • Response - CR 2 (67%) 3 (50%) - Other IWG response -- 1 (17%)
Long-Term Survival of Patients with Myelofibrosis Treated with Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Donato Michele L. , Pecora Andrew L. , Goldberg STuart L. , Hsu Jack W. , Siegel David S. , Goy Andre H. , Rowley Scott D. 
Published: October 2019

Myelofibrosis with myeloid metaplasia (MMM) is a disorder characterized by clonal myeloproliferation, ineffective erythropoiesis, extramedullary hematopoiesis and bone marrow fibrosis. Between 4/1998 and 1/2006, 17 patients with MMM were treated with allogeneic hematopoietic stem cell transplantation. Patients included 8 males and 9 females with a median age of 51.8 years (range 37–66). All patients were transfusion-dependent at the time of transplantation, 3 (18%) had abnormal cytogenetics and 4 (24%) had a prior splenectomy. Six patients (35%) received a reduced intensity conditioning regimen consisting of either fludarabine and busulfan (1 pt), fludarabine and low dose total body irradiation (1 pt), fludarabine and melphalan (3 pts) or fludarabine and cyclophosphamide (1 pt). Eleven patients (65%) received an ablative regimen of either busulfan and cyclophosphamide (10 pts) or cyclophosphamide and total body irradiation (1 pt). Patients with unrelated donors also received ATG as part of their conditioning. The hematopoietic cell source was peripheral blood in 12 pts (71%) and bone marrow in 5 pts (29%). Six pts (35%) received cells from an HLA-identical sibling, 1 pt (6%) had a 1 Antigen mismatch sibling and 10 pts (59%) received stem cells from an unrelated donor. Graft-versus-host disease prophylaxis consisted of tacrolimus in 15 pts (88%) and cyclosporine in 2 pts (12%).
Transplant-related mortality was 17% (sepsis 1 pt, VOD 1 pt, acute GVHD 1 pt) all occurring in the group treated with a myeloablative regimen. Of the remaining 14 patients, all achieved hematological recovery and transfusion independence. There were 2 relapses and one patient died of disease recurrence; both relapses occurred in the group receiving a reduced intensity regimen. The other 12 patients remain alive and in remission. The 6-year actuarial survival is 67% with no statistical difference between the groups who received a reduced intensity versus a myeloablative regimen.
In conclusion, Myelofibrosis with Myeloid Metaplasia can be successfully treated with allogeneic hematopoietic stem cell transplantation with a significant proportion of patients achieving long-term survival. Hematopoietic recovery can be achieved despite the presence of marrow fibrosis. The optimal preparative regimen remains to be determined with more relapses observed in the reduced intensity regimen group and a higher mortality seen in the myeloablative group. A risk-stratification strategy for regimen selection should be considered in future studies.
A Phase I Open Label Dose Escalation Study To Evaluate MEDI-507 in Patients with CD2-Positive T-Cell Lymphoma/Leukemia.
Authors: Casale Deborah A. , Bartlett Nancy L. , Hurd David D. , Foss Francine , Pro Barbara , Sokol Lubomir , Lieberman Ronald , Luz Hammershaimb 
Published: October 2019

Abstract
This ongoing multicenter study is a Phase I trial with MEDI-507 (Siplizumab) [a humanized IgG1k class monoclonal antibody that binds to the CD2 receptor on human T- and NK-cells] to determine the maximum tolerated dose (MTD) or the optimum biologic dose (OBD) in patients with relapsed/refractory CD2-positive T-cell lymphoma/leukemia [CD2-positive adult T-cell leukemia (ATL), cutaneous T-cell lymphoma (CTCL), peripheral T-cell lymphoma (PTCL), and large granular lymphocytic leukemia (LGL).] Open label 3+3 dose escalation was conducted in which patients receive bi-weekly infusions of MEDI-507 over 3 consecutive days at total doses of 0.7 mg/kg, 3.4 mg/kg or 4.8 mg/kg. Predose and serial MEDI-507 pharmacokinetics (ELISA) at Visit 2 and anti MEDI-507 antibodies (ELISA), peripheral blood total T-Cell and CD2-positive T-cells (flow cytometry), and C3 and C4 complement are obtained for each patient. Patients are followed for one year after their last dose of MEDI-507 for tumor assessment and CD2-positive T-cell recovery. 16 patients have been enrolled: 3 (0.7 mg/kg); 9 (3.4); 4 (4.8). Three additional patients were added to the 3.4 mg/kg cohort to replace patients who progressed early and one patient with tumor lysis syndrome. Diagnoses of the 16 patients: PTCL (9), CTCL (6) and NK-LGL (1). Frequent adverse events reported, to date, are infusion reaction (7/16) patients; hypertension (4/16); lymphopenia (4/16); fatigue (4/16) and leukopenia (3/16). Two DLT’s have been observed. The first, erythematous confluent dermatitis, occurred in one patient at 3.4 mg/kg and the cohort was expanded with eventual dose escalation. The second, pulmonary edema, occurred in one patient at 4.8 mg/kg and the cohort is currently being expanded without further DLT identified to date. Two responses have been observed, one PR in an NK-LGL patient at 3.4 mg/kg and one CR in a PTCL patient at 3.4 mg/kg. MEDI-507 has been well tolerated and shown anti-tumor activity. Dosing will change to weekly dosing with dose escalation (3+3) starting at 1.2 mg/kg as 0.8 mg/kg was seen to be safe in the single center study. MTD and/or OBD have not been determined.
RHAMM/CD168 Is a Novel Leukemia Associated Antigen with Prognostic Value for Patients with B-Cell Chronic Lymphocytic Leukemia.
Authors: Giannopoulos Krzysztof , Krober Alexander , Dmoszynska Anna , Rolinski Jacek , Dohner Hartmut , Stilgenbauer Stephan , Schmitt Michael 
Published: October 2019

Abstract
Background and Aims: Differential expression of molecules in patients with B-cell chronic lymphocytic leukemia (B-CLL) might define suitable targets for T cell based vaccines and/or antibody approaches.
Methods: We assessed the mRNA expression of the tumor associated antigen (TAA) RHAMM/CD168 defined earlier by serological analysis of cDNA expression libraries (SEREX) from leukemic cells.
Results: Peripheral blood mononuclear cells from 40 B-CLL patients and 20 healthy volunteers (HVs) were examined by quantitative RT-PCR. A leukemia-restricted expression of the antigen RHAMM/CD168 was observed in 39/40 B-CLL patients in contrast to the absence of its expression in HVs. To evaluate the immunogenicity of this novel LAA, mixed lymphocyte peptide cultures (MLPCs), followed by enzyme-linked immunosorbent spot (ELISPOT) and flow cytometry assays were performed to detect antigen-specific CD8+ T cells. RHAMM/CD168 specific responses by CD8+ HLA-A2/R3tetramer+CCR7-CD45RAhigh effector T cells were detected. As these CD8+ T cells contribute to the elimination of RHAMM+ CLL cells, we questioned whether expression of the antigen would be associated with a better survival. RHAMM/CD168 expression revealed to be higher in patients with unmutated IgVH status. RHAMM normalized against the housekeeping gene TATA binding protein (TBP), i.e. the RHAMM/TBP ratio was defined as a prognostic surrogate marker for B-CLL. B-CLL patients with a RHAMM/TBP ratio > 1.67 showed a significantly shorter treatment free survival (TFS) (Fig. 1). A tendency towards higher RHAMM/TBP expression ratios was observed in B-CLL cases with del11q.
Conclusion: RHAMM/CD168 is a novel LAA in B-CLL patients, an antigen correlating with the clinical course of the disease. Therefore, we consider RHAMM/CD168 an interesting target for immunotherapy in early stage B-CLL patients, especially with worse prognosis (IgVH unmutated).
Figure 1. Treatment-free-survival (TFS) according to RHAMM/TBP ratio Figure 1. Treatment-free-survival (TFS) according to RHAMM/TBP ratio
Integrating Prognostic Markers and Cellular Signaling Identifies More Chronic Lymphocytic Leukemia Patients with Adverse Prognosis.
Authors: Brennan Paul , Alghazal Suhair , Lloyd Naomi Price , Williams Janet E. , Griffiths Jeff D. , Rowntree Clare , Pratt Guy , Fegan Chris , Pepper Chris 
Published: October 2019

Abstract
In this study we devised an equation that provides a new way of identifying B-cell chronic lymphocytic leukemia (CLL) patients who will require early therapeutic intervention. The equation was developed by studying the biology of the disease with a focus on cell signal transduction. Using statistical analysis, we show that measured tyrosine phosphorylation can be described as a function of VH gene mutation status, and the expression of ZAP-70 and CD38 (r2 = 0.81, n = 49). Furthermore, we show that tyrosine phosphorylation has biological relevance as it sets a threshold for the activation of the transcription factor, NF-kB, resulting in the modulation of cell survival. Using the equation derived from our training dataset we calculated tyrosine phosphorylation in a cohort of 155 unselected CLL patient samples. The patient cohort was then divided into those with calculated tyrosine phosphorylation above or below the threshold required to activate NF-kB. Patients with high calculated tyrosine phosphorylation had a shorter time to first treatment. This method identified more patients at risk of requiring early treatment than VH gene mutation status, ZAP-70 or CD38. Furthermore, calculated tyrosine phosphorylation identifies patients that will require early intervention within the mutated VH gene cohort and even patients that have Stage A disease.