bluuebunny/crossref_metadata_2025_split huggingface dataset:
Motexafin Gadolinium, a Redox Mediator, Induces Apoptosis in Chronic Lymphocytic Leukemia B Cells and Is Additive or Synergistic with Fludarabine.
Authors: Naumovski Louie , Bone Nancy D. , Sirisawad Mint , Miller Richard , Jelinek Diane F. , Tschumper Renee C. , Kay Neil E. 
Published: April 2020

Abstract
Motexafin gadolinium (MGd, Xcytrin®) is a tumor-selective redox mediator that oxidizes intracellular reducing metabolites and protein thiols, thereby generating reactive oxygen species (ROS) through a process known as redox cycling. B-cell chronic lymphocytic leukemia (CLL) has elevated levels of the antioxidant ascorbate, and is known to be sensitive to redox stress. Since MGd can generate ROS through oxidation of ascorbate, we reasoned that MGd would be cytotoxic to CLL cells. Mononuclear cells were isolated from the blood of CLL patients (n=15) characterized by Rai Stage, CD38, FISH and immunoglobulin heavy chain variable (Ig VH) mutational status. CLL cells were incubated in vitro over 2–5 days with MGd at concentrations ranging from 0–50 uM. Cytotoxicity was assessed by flow cytometry using annexin V binding/propidium iodide uptake. Accumulation of MGd in CLL cells was demonstrated by flow cytometry using drug fluorescence (&gt;650 nm). MGd-induced cytotoxicity was found to be dose-dependent over a range of 2.5 to 50 uM in cells from 11 of 15 CLL patients. CLL cells from 11/15 patients treated with 50 uM MGd and 4/15 patients treated with 10 uM MGd had cytotoxicity levels at least 2 fold above baseline. MGd uptake was similar in sensitive and resistant CLL cells, implying that mechanisms other than uptake are responsible for the differential responses. CLL cells that were sensitive to MGd treatment showed an increase in caspase-3 activity and cleavage of the caspase substrate PARP was found by immunoblot suggesting that MGd induced apoptosis. The induction of apoptosis was found to be exclusive of Rai stage, CLL FISH defects, CD38 expression or Ig VH mutational status. Cells from 5 CLL patients were studied for drug synergy using seven concentrations of MGd (7.5 – 50 uM) and the active moiety F-ara-A of fludarabine (0.25 –2.0 uM). The cells were treated with each drug individually or both drugs for 48 hours prior to analysis for cytotoxicity. MGd was found to exhibit additive activity (N=4) or synergistic (N=1) effects on CLL B cytotoxicity when combined with fludarabine. Thus, our pre-clinical results reveal that MGd is cytotoxic to CLL cells in vitro through induction of apoptosis. Furthermore, MGd shows additive or synergistic activity when combined with fludarabine. These results support the evaluation of MGd in ongoing CLL clinical trials utilizing this agent alone or in combination with fludarabine.
Effect of 2-Deoxyglucose, a Glycolytic Inhibitor, on the ATP Levels and Viability of B Lymphocytes from Subjects with Chronic Lymphocytic Leukemia (CLL).
Authors: Tidmarsh George F. , Tanner Laura I. , O’Connor Julie , Eng Clarence , Mordec Kasia 
Published: April 2020

Abstract
One hallmark of tumor cells is a high rate of glycolysis, which underlies the utility of the glucose analog, 2-[18]fluoro-2-deoxyglucose for PET imaging of solid tumors. The glucose analog, 2-deoxyglucose (2DG), inhibits glycolysis, and is predicted to decrease the viability of tumor cells, which are primarily dependent on the glycolytic pathway for ATP generation. We have recently shown that 2DG enhances the efficacy of either adriamycin or paclitaxel in delaying tumor growth in mouse xenograft cancer models (Maschek et al, Cancer Res 64 (2004): 31). We are currently conducting a phase 1 clinical trial to examine the safety of a combination administration of 2DG and Taxotere in patients with solid tumors. The goal of the current study is to measure the extent to which 2DG inhibits energy production and, consequently, reduces viability of leukemic cells. Heparinized blood was obtained from four subjects with a confirmed diagnosis of CLL (peripheral lymphocyte blood count ranged from 5,000–24,000/mm3), with an additional sample drawn from each subject two weeks later for replicate determinations. The mononuclear cell fraction (predominantly B cells) was isolated using Histopaque. Subsequent treatment of isolated CLL cells with 2DG (164 μg/ml) at 37°C (in the presence of 0.5 mM glucose) resulted in a 20–30% decrease in cellular ATP levels after 1 hr (compared with untreated cells) and a 60–70% decrease in ATP levels after 24 hrs. In the presence of 5 mM glucose, incubation of CLL cells with 2DG elicited a 5–10% decrease in ATP at 1 hr and 15–20% reduction in ATP levels at 24 hrs. The effect of 2DG on cell viability was assessed using Alamar Blue. Treatment of CLL cells with 2DG (164 μg/ml, in the presence of 0.5 mM glucose) resulted in a 50% loss in cell viability at 24 hrs (compared with untreated cells at the same time point), and a 70% loss at 48 hrs. In the presence of 5 mM glucose, the effect of 2DG was more modest. At 24 hrs, there was a 20% loss of cell viability and a 30–50% loss at 48 hrs. In conclusion, these results demonstrate that in vitro treatment of CLL B cells with 2DG decreases cellular ATP levels, ultimately resulting in a loss of cell viability. Updated results with samples from additional CLL subjects will be presented, together with an assessment of the therapeutic potential of 2DG for the treatment of CLL.
Treatment of Patients with Chronic Lymphoproliferative Disorders (CLPD) with Fludarabine, Cyclophosphamide, and Rituximab (FCR) Combination.
Authors: Galanopoulos Athanasios G. , Tsakiridou Anastasia , Michalis Eurydiki , Marinakis Theodoros , Gortzolidis George , Zomas Athanasios , Tsourveloudis Ioannis , Tsirogianni Maria , Papadhimitriou Stephanos , Kritikou-Griva Elpiniki , Anagnostopoulos Nikolaos I. 
Published: April 2020

Abstract
The FCR combination in a phase III study (Blood 102, abstract #373, p110a,2003) has shown promising results in the treatment of chronic lymphocytic leukemia (CLL) patients leading in increase of proportion of complete remissions and improved survival. Based on these encouraging results we tried to study the efficacy and safety of the FCR combination in patients of our Haematologic Unit. Seventeen patients, 8 males and 9 females with a median age of 69,5 years old, with relapsed/refractory or de novo CLPD ( 9 CLL and 8 NHL patients) were enrolled in this study between February 2002 and August 2004. Fifty percent of CLL patients had Rai stage I/II and the rest 50% had Rai stage III/IV disease. Four NHL patients had also Ann Arbor stage I/II and the rest four Ann Arbor stage III/IV disease. They were treated with Rituximab 375 mg/m2 on day1 in combination with Fludarabine and Cyclophosphamide ( 25 mg/m2 and 250 mg/m2 respectively) for days 2 to 4, every 4 weeks, for 6 consecutive cycles. Nine patients had a history of a prior unsuccessful treatment. Overall, 14 out of 17 evaluable patients were responsive to the treatment [12 patients complete response (CR) and 2 patients partial response (PR), overall response rate (ORR) 82%]. The remaining 3 patients did not show any response (NR), 2 progressive and 1 stable disease. Hematological toxicity was acceptable ( grade 2–3 neutropenia in 6/17 patients, grade 2–3 anemia and thrombocytopenia in 2/17 patients). There were no septic episodes except one case with neutropenic fever. There were no adverse events like nausea or vomiting except one patient with a serious anaphylactic reaction due to Rituximab administration. Three CLL patients died because of stable or progressive disease. In conclusion, this preliminary report suggests that the FCR regimen is an effective and safe treatment for CLPD patients, achieving higher CR rates than previous treatments. A longer follow up of a larger number of patients is required to confirm an improved survival in these patients.
FLT3/ITD Expression Increases Expansion, Survival and Entry into Cell Cycle of Human Hematopoietic Stem Cells.
Authors: Li Li , Piloto Obdulio , Kim Kyu-Tae , Ye Zhaohui , Nguyen Bao , Yu Xiaobing , Levis Mark , Cheng Linzhao , Small Donald 
Published: April 2020

Abstract
FMS-like tyrosine kinase-3 (FLT3) is a Class III receptor tyrosine kinase that is important for normal hematopoiesis. Activating mutations of FLT3 by internal tandem duplications (ITDs) in the juxtamembrane region are the most common molecular aberrations found in acute myeloid leukemia (AML). The contributions of FLT3 activating mutations in the leukemic transformation of normal human hematopoietic stem/progenitor cells (HSCs) have not yet been fully elucidated. In this study, using a single lentiviral vector containing two promoters, we achieved consistent and efficient coexpression of FLT3/ITD and green fluorescent protein (GFP) in transduced human CD34+ HSCs. When cultured in medium containing SCF, TPO and FLT3 ligand (FL), FLT3/ITD-transduced cells survived with enhanced self-renewal and survival potential, which was not affected by the withdrawal of FL. These cells retained a surface immunophenotype typical of HSCs (CD34+CD38−). Compared to cells transduced with a vector expressing GFP alone, FLT3/ITD-transduced HSCs had a higher fraction of cells in cell cycle. Clonogenic assays showed that FLT3/ITD-transduced HSCs produced fewer CFU-GM, implying that they were at least partially blocked in their ability to differentiate along the myeloid lineage. FLT3/ITD-transduced HSCs were more sensitive to the induction of cytotoxicity by CEP-701, a selective FLT3 inhibitor. In the FLT3/ITD-transduced HSCs, we detected increased expression of Pim-1, a serine/threonine kinase with an important role in cell survival, proliferation and differentiation, c-myc, a transcription factor involved in cell proliferation and cell cycle regulation, and Cyclin D3, a key factor in cell cycle regulation, each of which may contribute to the altered genetic program instituted by FLT3/ITD signaling.
These results together indicate that FLT3/ITD mutations may contribute to leukemic transformation of normal HSCs by prolonging survival, promoting proliferation, and blocking differentiation. CEP-701 may act as a potent agent for AML stem cells harboring FLT3/ITD mutations.
Cyclin D1 Immunohistochemistry Identifies a Subset of Plasma Cell Myeloma with Superior Overall Survival.
Authors: Cook James R. , Hsi Eric D. , Tubbs Raymond R. , Worley Sarah , Hussein Mohamad A. 
Published: April 2020

Abstract
The expression of Cyclin D1 is dysregulated in approximately half of cases of plasma cell myeloma due to translocations, aneusomy, or other abnormalities. Recent studies using quantitative mRNA analysis have suggested that increased Cyclin D1 mRNA expression is associated with a favorable prognosis. Previous attempts to examine the significance of cyclin D1 protein expression by immunohistochemistry have been hampered by the use of antibodies with weak staining and high background. In this study, we employ a newly available, commercial antibody that gives superior staining in B5 fixed tissues. We performed immunohistochemistry for Cyclin D1 on bone marrow core biopsies from a series of 44 newly diagnosed plasma cell myeloma patients who were uniformly treated on a Phase II study of rituxan, melphalan and prednisone. 22 patients (50%) were positive for Cyclin D1, defined as any plasma cells with positive nuclear staining. Cyclin D1 positive and negative cases displayed no significant differences in the initial levels of β2m (3.6±0.5 mg/L vs. 3.5±0.5 mg/L, p=0.860), number of bone marrow plasma cells (63±5.5% vs. 47±6.2%, p=0.063), or proportion of cases classified as SWOG stage 3-4 (2 of 22 (9%) vs. 5 of 22 (23%), p=0.412). The cyclin D1 positive cases displayed a superior overall survival with an estimated 3-year survival of 95% for Cyclin D1 positive cases versus 56% for Cyclin D1 negative cases (p=0.032). The cyclin D1 positive cases also displayed a trend towards better progression-free survival (median progression free survival of 15.7 months for Cyclin D1 positive versus 12.8 months for Cyclin D1 negative, p=0.13). In a Cox proportional hazards regression model, used to compare the effect of Cyclin D1 protein expression on overall survival time while adjusting for stage, the Cyclin D1 positive patients continued to show a strong trend to better overall survival (p=0.062). This study demonstrates that cyclin D1 immunohistochemistry, which could be readily performed in most pathology laboratories, is capable of identifying a subset of plasma cell myeloma with a favorable survival. Additional studies are ongoing to determine if these results can be generalized to other forms of therapy. If confirmed, routine cyclin D1 immunohistochemistry at the time of diagnosis may offer important prognostic information that could identify lower risk patients for whom less intensive therapies might be appropriate.
Ploidy, as Detected by Fluorescence In Situ Hybridization, Defines Different Subgroups in Multiple Myeloma.
Authors: Avet-Loiseau Herve , Wuilleme Soraya , Robillard Nelly , Lode Laurence , Magrangeas Florence , Beris Helen , Harousseau Jean-Luc , Proffitt John , Minvielle Stephane 
Published: April 2020

Abstract
Ploidy appears as an important parameter in both the biology and the clinical evolution of multiple myeloma. However, its evaluation requires either a successful karyotyping (obtained in 30% of the patients), or a DNA index calculation by flow cytometry (not routinely performed in myeloma). We validated a novel method for the detection of hyperdiploidy based on interphase fluorescence in situ hybridization (FISH) that can be utilized on almost all patients. We selected a 3-color probe set composed of probes specific for chromosomes 5, 9, and 15. Two hundred and five patients with myeloma were analyzed by flow cytometry (DNA Index) and FISH, and results obtained by both techniques were then compared. As shown in the table, FISH was very specific and sensitive for the detection of hyperdiploidy. Only 6 of the 104 hyperdiploid patients, all with a low hyperdiploidy (DNA Index &lt;1.10), were classified as diploid by FISH. Extended studies looking for t(4;14), t(11;14), and del(13q14) showed that most recurrent 14q32 translocations occur in non-hyperdiploid clones, and that deletions of chromosome 13 were less frequently observed in hyperdiploid clones (48% vs 66%). These data confirmed that chromosomal abnormalities are not randomly distributed in myeloma and that FISH allows analysis of all the major chromosomal abnormalities. Further large studies are ongoing to evaluate the prognostic value of ploidy in myeloma and to correlate the true prognostic value of all chromosomal abnormalities.
Diploid by FISH Hyperdiploid by FISH Tetraploid by FISH t(11;14) t(4;14) del(13q14) DNA index &lt; 1 7 3 2 3 DNA index=1–1.04 91 42 17 61 DNA index=1.05–1.5 6 98 7 7 50 DNA index &gt;1.6 3 1 2
Expression of TRAIL Receptors on the Multiple Myeloma Cells.
Authors: Li Juan , Li Junhe , Luo Shaokai , Zhao Yin 
Published: April 2020

Abstract
Objective To study the different expression of death receptors and decoy receptors on mononuclear cells from patients with multiple myeloma and myeloma cell line KM3 and compare the different expression of TRAIL receptors after chemotherapy or exposure to doxorubicin, to explore the mechanisms by which TRAIL selectively kills tumor cells.
Methods Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and flow cytometry was used to investigate the expression of four receptors on mononuclear cells in 23 multiple myeloma patients and myeloma cell line KM3 and 15 controls, we furthermore compared the changes of expression mode after chemotherapy and incubation of KM3 cell with sub-clinical concentration of Doxorubicin.
Results There finds only DR4 and DR5 on KM3 cell line without the expression of DcR1 and DcR2. Expression of DR4 and DR5 on mononuclear cells of MM patients is higher than that of controls (P&lt;0.05), but DcR1 and DcR2 expression was lower than that of controls (P&lt;0.05), after chemotherapy and exposure to Doxorubicin, the expression of DR5 on MM cells was up-regulated (P&lt;0.05)
Conclusions The expression of four receptors on myeloma cells and normal controls was significantly different, which might account for the selective killing effect of TRAIL on MM cells. DR5 was up-regulated on KM3 when incubating with Doxorubicin and after chemotherapy which suggests chemotherapy agents might enhance the apopotosis of MM cells through up-regulating of DR5 receptor.
Frequent Detection of Minimal Residual Disease (MRD) in Autografts from Patients with Multiple Myeloma during Remission Using a New Culture Method.
Authors: Cohen Yossi , Hardan Yzhar , Nagler Arnon , Zipori Dov 
Published: April 2020

Abstract
Current treatment of multiple myeloma includes autologous stem cell transplantation. However, it is unknown at the moment what is the extent of graft contamination with clonotypic myeloma cells. In order to evaluate the extent of residual contamination of the graft with myeloma cells, we used our new myeloma cell culture and expansion method developed in the Weizmann Institute of Science for the detection of MRD. We observed readily growing residual myeloma cells in 6 of seven cases, confirmed by clonal markers (FACS, PCR and FISH). However, there was some variability in the pattern of growth; one case of plasma cell leukemia and two cases with t(4;14) showed earlier and more pronounced growth, whereas one case with systemic amyloidosis and another case with MGUS failed to grow in this culture. We are currently arranging a multicenter study for further assessment of these findings and aim to answer the question whether the culture can distinguish between multiple myeloma and other plasma cell dyscrasias. Another goal is to correlate the pattern of in vitro growth of multiple myeloma cells, with clinical and chromosomal characteristics.
Figure Figure
Low Incidence of Deletions of ATM (ataxia-telangiectasia mutated) Gene in Patients (pts) with Multiple Myeloma (MM) as Detected by FISH.
Authors: Chanan-Khan Asher Alban , Miller Kena , McCarthy Philip , Panasiewicz Joanna M. , Block AnneMarie W. 
Published: April 2020

Abstract
Loss of function of ATM gene is associated with higher incidence of leukemia and lymphoma. ATM localized to 11q22.3-q23.1, is a tumor suppressor gene and plays an important role in cell cycle arrest, DNA repair mechanisms, and apoptosis. The incidence of deletion of ATM in MM remains unknown. Using a new commercially available fluorescently labeled DNA probe, we investigated the incidence of homozygous and heterozygous deletions of the ATM gene locus to explore its potential role in the pathogenesis of multiple myeloma (MM). Fifty-three MM pts were evaluated by fluorescence in situ hybridization (FISH) using the Vysis, Inc. LSI ATM (11q22.3) probe to assess the status of the ATM gene. LSI ATM (11q22.3) is a ~500 kb probe that hybridizes to the 11q22.3 region of chromosome 11 and spans the entire ~184 kb ataxia telangiectasia mutated (ATM) gene and several others. Probe copy number was assessed in each patient after analyzing at least 200 interphase nuclei. Only pts with&gt; 10% bone marrow involvements with malignant plasma cells were analyzed. Heterozygous ATM deletion was noted in 2/53 (3.7%) patients. These patients had 82% and 20% involvement of bone marrow (BM) with malignant plasma cells. Twenty-eight of 53 (%) patients showed an extra signal suggestive of trisomy 11, while no abnormality (2 signals) was noted in 23/53 (%) pts. There was no significant difference in the stage, B2M or extent of marrow involvement among patient with or without aberration of ATM gene. Up to 15–20% of the pts with B-cell lymphoproliferative disorders (CLL, mantle cell and other NHL) are reported to have ATM deletion suggesting a possible role of ATM protein in the pathogenesis of at least a subset of these disorders. Even though MM is a B-cell disorder, in our observation the prevalence of ATM deletion in these pts is uncommon and thus unlikely to be a major factor in the pathogenesis of MM.
Rituximab in Anti-Mag Associated Neuropathy : A Monocentric Experience.
Authors: Delarue Richard , Deau Benedicte , Alyanakian Marie-Alexandra , Varet Bruno R. , Hermine Olivier 
Published: April 2020

Abstract
Anti-MAG associated neuropathy is a rare disorder characterized by a sensory-motor polyneuropathy in the context of monoclonal IgM, which activity is directed against myelin-associated glycoprotein on the surface of myelin sheaths. Treatments are usually ineffective. Most common therapy is based on alkylating agents such as Chlorambucil, which however only rarely improve neurological tests nor induce disappearance of monoclonal component. More recently, the benefit of monoclonal anti-CD20 antibody Rituximab was reported in case reports. Thus, we have performed since 2002 a prospective pilot study to assess the benefit of Rituximab in anti-MAG associated neuropathy. We report here the first four successive patients treated.
Patients and Methods : inclusion criteria were as follow : peripheral sensory-motor polyneuropathy with clinical and electrophysiological symptoms compatible with anti-MAG associated neuropathy; serum monoclonal IgM; serum anti-MAG reactivity; absence of other cause of peripheral neuropathy such as amyloidosis; absence of underlying condition contra-indicating Rituximab therapy. Patients received 4 courses of Rituximab (375 mg/m2) with a 7 days interval between two infusions. Usual premedication with corticosteroids and paracetamol were administered. Evaluation of clinical, biological and immunochemical efficiency was performed every three months.
Results : characteristics of the first four patients treated are as follow : 3 males and 1 female; median age : 65 years (57 – 87); median time between onset of symptoms and Rituximab therapy : 4 years (3 – 6). Previous treatments were Chlorambucil (3/4), corticosteroids (2/4), intravenous Ig (1/4), anthracyclin-containing regimen (1/4), plasmapheresis (1/4). All were ineffective. Rituximab was well tolerated by all patients. Median follow-up after treatment is 2 years (1 – 3). None of the patients exhibit improvement of clinical neurological symptoms. Electrophysiological evaluation show only a mild improvement in 1 patient. Moreover, no decrease in monoclonal component nor disappearance of anti-MAG reactivity of serum were observed in any cases. Interestingly, one patient exhibited later neurological and biological improvement after high dose Melphalan followed by autologous stem cell transplantation.
Conclusion : Rituximab monotherapy does not seem to improve neurological outcome of patients with anti-MAG polyneuropathy as well as is unable to clear serum from monoclonal IgM. Therefore, new approaches are needed, for example with high dose chemotherapy with stem cell support in selected patients.