bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical Significance of Free Circulating CD3 and CD5 in Patients with Chronic Lymphocytic Leukemia.
Authors: Hodge Menna , O’Brien Susan , Abdool Adam , Keating Michael , Jilani Iman , Kantarjian Hagop , Ferrajoli Alessandra , Estrov Zeev , Giles Francis , Albitar Maher 
Published: April 2020

Abstract
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CD5, a transmembrane protein expressed in T-cells, few B-cells, and chronic lymphocytic leukemia (CLL) B-cells, is the ligand for CD72 and may play a role in B-cell-T-cell communication. CD5 is part of the T-cell receptor (TCR)-CD3 complex in T-cells as well as the B-cell receptor (BCR) complex and serves as substrate for induction of tyrosine kinase activity. Since leukemic cells have high turnover and pour their protein, RNA, and DNA into the circulation, we speculated that free circulating CD3 (cCD3) and CD5 (cCD5) could be detected in the plasma of patients with CLL. We have developed a bead-based sandwich immunoassay to measure cCD3 and cCD5 in the plasma. Using this assay, we assessed the value of cCD5 measurement, alone and after normalization to cCD3 levels, as a tumor marker in CLL. Plasma levels of cCD3 and cCD5 were measured in 85 patients with CLL and 51 normal control subjects. cCD3 and cCD5 levels were significantly higher in patients with CLL (median, 7,465 and 55,806 U/μl, respectively) than in normal control subjects (median, 830 and 1,671 U/μl, respectively). Patients with CLL had significantly higher cCD5:cCD3 ratios (median, 5.28; range, 0–161) than did normal controls (median, 1.70; range, 0–8.06) (P &lt;0.0001). Levels of cCD5, but not cCD3, correlated positively with WBC count, β2-microglobulin level, splenomegaly, and Rai stage (all P &lt;0.01). The cCD5:cCD3 ratio also correlated with Rai stage (P = 0.04) and β2-microglobulin level (P = 0.03). cCD5 levels and the cCD5:cCD3 ratio both correlated with survival (P = 0.03). These findings confirm that free circulating surface markers can be detected in the circulation of patients with CLL, most likely reflect the tumor load, and can be used as tumor markers. The biological and therapeutic relevance of these free circulating proteins should be considered in pharmacokinetic and pharmacodynamic studies.
Valproic Acid Induces Apoptosis in Chronic Lymphocytic Leukemia Cells through Activation of the Death Receptor Pathway and Potentiates TRAIL Response.
Authors: Lagneaux Laurence , Gillet Nicolas , Delforge Alain , Dejeneffe Marielle , Stamatopoulos Basile , Massy Martine , Meuleman Nathalie , Kentos Alain , Willems Luc , Bron Dominique 
Published: April 2020

Abstract
Background: The anti-leukemic in vitro activity of valproic acid (VPA), a commonly used antiepileptic agent, was tested on lymphocytes derived from 40 patients with chronic lymphocytic leukemia (CLL) (Binet stage A=34, B=3, C=3). These patients had not been previously treated or remained untreated for the previous 6 months. Combined analysis of ZAP-70, CD38 and IgVH mutational status was performed for each patient.
Methods: Mononuclear cells were incubated with VPA at 1, 5 and 10 mM for 24 hours. Cell viability was assessed by trypan blue exclusion assay, apoptosis by annexin V/propidium iodide(PI) labelling and PI staining after cell permeabilisation. Caspase activation was studied by flow cytometry analysis after cell treatment with selective caspase inhibitors.
Results: Exposure of CLL cells to VPA resulted in dose-dependent cytotoxicity and apoptosis in all CLL patients tested. VPA-treatment induced apoptotic changes in CLL cells including phosphatidylserine (PS) externalisation and DNA fragmentation. The mean apoptotic rate was similar between IgVH mutated and unmutated patients or ZAP-70+/ZAP-70- cases. VPA induced apoptosis by the extrinsic pathway involving engagement of the caspase-8 dependent cascade. Although CLL cells are commonly resistant to death receptor-induced apoptosis, VPA increased significantly the sensitivity of leukemic cells to TRAIL (tumor necrosis factor α-related apoptosis-inducing ligand). In addition, VPA overcomed the prosurvival effects of bone marrow stromal cells.
Conclusions: These data indicate that VPA, at the pharmacological concentration of 1 mM, is a potent inducer of apoptosis in CLL and should be further explored as a single agent. Also the combination of VPA and TRAIL may be a promising approach in the treatment of CLL.
Transcriptosome and Serum Cytokine Profiling of B-CLL Identifies Oncogenic Pathways in Pathogenesis and Survival.
Authors: Choi James , Garewal Harinder , Riley Christopher , Cooke Laurence , Klinkhammer Thomas , Maddipati Sirisha , Sud Rohit , Spier Catherine , Mahadevan Daruka 
Published: April 2020

Abstract
Background: B-cell chronic lymphocytic leukemia (B-CLL) the most common leukemia in the Western world accounts for 25% of all newly diagnosed leukemias. Despite new therapeutic advances, B-CLL is currently not curable. Potential oncogenic signaling pathways in B-CLL require elucidation. We identified ROR-1 receptor tyrosine kinase (RTK) in the WNT/Planar Cell Polarity non-canonical pathway as a possible mechanism of oncogenesis by gene expression profiling (GEP).
Methods: Mononuclear cells from 8 low stage CLL patients were obtained through an IRB approved protocol and analyzed utilizing the HG-U133A 2.0 Affymetrix array (~18,400 transcripts, 22,000 probe sets) after isolating and purifying total RNA (Qiagen, RNAeasy). The control RNA samples were isolated from 8 normal volunteer peripheral blood (PB) B-cells (AllCell, CA) and a normal reactive lymph node. Tumor lineage was confirmed by immunohistochemistry (IHC). Quantitative real time RT-PCR was performed on 20 selected genes to validate the microarray GEP. Twenty one B-CLL patients (includes the 8 patients from the GEP) were evaluated by RT-PCR for several key components of oncogenic signaling pathways based on the GEP with gene specific probes. A serum cytokine profile (120 cytokines) on 12 CLL patients (8 for which GEP is available and 4 additional patients) and 4 normal volunteers were performed for identification of a signature for diagnostic and prognostic value.
Results: Data are represented as “robust” increases or decreases of relative gene expression common to 8 patients. ROR-2, a close member of ROR-1, has been shown to bind Wnt-5A via the cysteine rich domain (CRD) and to activate the JNK signaling pathway. Our findings identify over-expression of members of the non-canonical WNT/PCP- ROR-1- signaling pathway genes that were validated by RT-PCR. A serum cytokine profile of 12 patients provides a signature that may be useful in CLL diagnosis and prognosis.
Conclusions: GEP identified WNT/PCP-ROR-1 as key components of an autocrine pathway that helps B-CLL avoid apoptosis. Several serum cytokines are elevated and require validation as potential diagnostic and prognostic markers. GEP of B-CLL in combination with quantitative real time RT-PCR has identified several novel targets for therapy. The identification of ROR-1 RTK has led to the development of a molecular target for future therapeutic application. Several lead compounds have been identified and are being evaluated as potential therapies in B-CLL.
Ubiquitously Expressed Micro RNA miR-16 Is Not Mutated and Not Differentially Expressed in Patients with B-CLL.
Authors: Landgraf Pablo , Fuchs Uta , Novosel Astrid , Fulci Valerio , Schoch Claudia , Haferlach Torsten , Buhmann Raymund , Tuschl Thomas , Borkhardt Arndt 
Published: April 2020

Abstract
MicroRNAs (miRNAs) are a recent discovered class of conserved 21–23 nucleotide short non-coding RNAs regulating protein levels by translational inhibition and mRNA destabilization and are involved in a variety of processes including cell differentiation and apoptosis. About 68% of miRNAs are tissue specifically expressed and great attention was paid to this group of miRNAs. To assess which miRNAs are ubiquitously expressed we investigated 170 human, 68 mouse and 16 rat libraries from 26 different organ systems and cell types by cloning and sequencing a total of 332,070 small RNA sequences comprising 220,147 miRNA sequences. 39 human miRNA clusters are transcribed in at least 70% of tissues and cell types. Interestingly the only miRNA cluster detected in nearly all libraries including those of embryonic origin and 8 B-CLL patient samples are the mir-16 clusters comprising mir-16-1 with mir-15a on chr.13q14 and mir-16-2 with mir-15b on chr.3q26. Earlier reports linked a lower abundance of the mature miR-16 to loss of heterozygosity (LOH) of chr.13q14 and mutations in the precursor were implicated in B-CLL in those reports. We therefore extended our investigation of B-CLL samples. We did not find any mutation in the miR-15a-miR-16-1 gene in all patients examined (64 patients). There were also no mutations in 35 healthy controls.
Furthermore 37 of 39 patients had detectable levels of miR-16 in quantitative Northern blot analysis. Only one of the two patients with undetectable miR-16 had a positive LOH status of the 13q14 locus in 90% of cells examined by FISH while the other one had no chromosomal aberrations or deletions. Additionally, the expression level of miR-16 is not correlated with the del13q14 LOH status in our study (P = 0.43 in double sided Fisher’s exact test). We also investigated the stability and processing of a reported construct bearing a C-&gt;T mutation 7 nt 3′ of the mature miRNA in 293 cell culture. This mutation has been reported in two B-CLL patients. We could not detect any difference in miR-16 expression compared to the wild type construct. Another ubiquitously expressed miRNA miR-21 is less stably expressed and is not detectable in 7 out of 39 patient samples by quantitative Northern blotting. We conclude that the absence of miR-16 in patients with B-CLL is rare and is not due to mutations in the miR-15a-miR-16-1 gene cluster in the patients examined. Furthermore miR-16 seems not to be affected by gene dosis effects like other genes involved in the LOH of 13q14 maybe because of compensatory up regulation of the other miR-16 cluster.
Dipeptidyl Peptidase 2 - A Pro-Survival Molecule in Chronic Lymphocytic Leukemia.
Authors: Danilov Alexey V. , Danilova Olga V. , Klein Andreas K. , Huber Brigitte T. 
Published: April 2020

Abstract
Chronic lymphocytic leukemia (CLL) is a unique malignancy characterized by persistent accumulation of quiescent B-cells. Disruption of survival pathways leads to apoptosis and renders CLL cells sensitive to chemotherapeutic agents. As clinical outcomes in CLL are heterogeneous, it is important to better predict prognosis in each individual case of the disease. In this respect, detection of somatic mutations in the immunoglobulin variable heavy chain (IgVH) genes has become the gold standard. Overexpression of ZAP-70 (Zeta-chain associated protein kinase - 70 kDa) by CLL cells correlates with unmutated IgVH genes and predicts poor outcome in B-CLL. Dipeptidyl Peptidase 2 (DPP2) is a newly discovered serine protease which maintains lymphocytes in a quiescent state (G0). In this study we investigated whether DPP2 is involved in cell cycle control in B-CLL. 38 patients with B-CLL and 20 healthy controls were included in the study. Median age of CLL patients was 67 years. Median time from diagnosis to enrollment in the study was 102 months. 21 patients (55.3%) received treatment before enrollment in the study. Standard Ficoll-Hypaque techniques were used to isolate peripheral blood mononuclear cells (PBMC) from healthy donors and CLL patients. CLL cells were isolated to &gt;98% purity by means of a MoFlo using CD19−specific antibodies. Cells were treated with Val-boro-Pro, a pan-inhibitor of DPP, or with DPP2-specific inhibitor (DSI), incubated for 16 hours and stained with propidium iodide and Annexin V. Expression of CD38 was assessed by flow cytometry, DPP2 and ZAP-70 - by real-time reverse-transcription polymerase chain reaction, Bcl-2 and p27 - by western blot analysis.
We report that CLL B-cells expressed higher levels of DPP2 mRNA than normal B-cells. Inhibition of DPP2 in PBMC with VbP or DSI resulted in caspase-dependent apoptosis. In individuals with CLL, death of B-lymphocytes (and rescue by caspase inhibitors) was observed in 22 cases (57.9%). In the remaining 16 cases (42.1%) malignant B-cells did not undergo apoptosis upon inhibition of DPP2 with either VbP or DSI. We found that CLL cells resistant to DPP2 inhibition-induced apoptosis expressed higher levels of ZAP-70. Meanwhile, protein levels of p27 (cell cycle inhibitor) were decreased in resistant CLL. These patients exhibited worse disease prognosis such as shorter treatment-free period (p&lt;0.001), frequent episodes of treatment failure and faster disease progression. Between the two groups, we identified no differences in expression of Bcl-2. CLL B-cells express higher levels of DPP2, a serine protease involved in maintenance of G0 which may serve as a survival factor in CLL B-cells. Resistance vs. susceptibility to DPP2 inhibition-induced apoptosis can be employed as a prognostic factor in CLL.
Therapy with Fludarabine, Cyclophosphamide and Rituximab (FCR) for Relapsed or Untreated Progressive Chronic Lymphocytic Leukemia (CLL). A Single Center Experience.
Authors: Pavlovsky Carolina , Pavlovsky Santiago , Pardo Maria L. , Sapia Sandra , Monreal Mariela , Corrado Claudia , Pavlovsky Miguel A. , Fernandez Isolda , Juni Mariana 
Published: April 2020

Abstract
Purpose: To evaluate the efficacy of FCR in improving complete remission (CR), disease-free survival (DFS) and overall survival (OS) rates in patients previously treated with chlorambucil-prednisolone and untreated CLL patients.
Patients and methods: A total of 45 CLL patients started FCR. Forty-one patients completed treatment: 16 following previous relapse and 25 previously untreated with progressive disease. Four patients are still receiving treatment. Median patient age was 63 years (range 34–88 years). Binet’s stage were: A: 8%, B: 34% and C: 58%. CD38 expression was positive (&gt; 7% of cells) in 56% of patients and negative in 44%. FCR consisted of: fludarabine (25 mg/m2/day × 3); cyclophosphamide (250 mg/m2/day × 3) and rituximab (375 mg/m2/day × 1), all given intravenously, every 4 weeks for 4–6 cycles. CR was defined by CLL/NCI-WG criteria. Minimal residual disease (MRD) negativity was &lt; 1% CD19, CD5 positive cells in peripheral blood and bone marrow. Results: The results of this study are detailed in Table 1. To summarise: CR: 69%; nodular partial remission (PR): 15%; PR: 7% and stable disease: 2%. Grade 3–4 neutropenia occurred in 33% of patients and 34% required hospitalization due to infections.
Table 1: Patient details following FCR treatment Patients % CR % MRD negative % DFS 36 mo P % OS 36 mo P Untreated 25 87 96 90 95 Treated 16 53 87 75 0.10 89 0.90 Total 41 74 92 83 93 CD38 Positive 23 71 90 71 0.02 87 0.27 CD38 Negative 18 76 94 100 100
Conclusions: FCR induces a high CR (74%) and DFS (83%) rate and increases MRD negativity (92%). Significantly higher DFS rates at 36 months were observed in patients who were CD38 negative vs. CD38 positive (100 % vs. 71% P=0.02).
Constitutive Nuclear p65 NF-kB Expression Predicts for Spontaneous Apoptosis and In Vitro Sensitivity to Fludarabine in CLL Cells.
Authors: Hewamana Saman , Rowntree Clare , Lin Thet Thet , Pepper Chris , Brennan Paul , Fegan Chris 
Published: April 2020

Abstract
B-cell chronic lymphocytic leukemia (CLL) is characterized by the accumulation of monoclonal B-cells in the blood, secondary lymphoid tissues, and bone marrow. The leukemia cells primarily are arrested in the G0/G1 phase of the cell cycle and appear resistant to programmed cell death. Despite their apparent longevity in vivo, CLL cells typically undergo spontaneous apoptosis in vitro although there is considerable heterogeneity in the timing and extent of this process. In this study we investigated the degree of this apoptosis in vitro, and its association with constitutive nuclear p65 NF-kB expression (n = 25). Apoptosis was measured by annexin V / propidium iodide labeling using flow cytometry and nuclear NF-kB expression was evaluated using electrophoretic mobility shift assay and quantified using a p65 ELISA. In contrast to previous reports, we found constitutive NF-kB activity varied in our patient cohort with a trend towards higher expression in samples with unmutated VH genes, high CD38 expression and high ZAP-70 expression. Importantly, we demonstrated an inverse correlation between the percentage of Annexin V-positive cells and nuclear p65 NF-kB expression (r2 = − 0.21; P = 0.02) suggesting that high levels of p65 NF-kB are cytoprotective in CLL cells. We then investigated the relationship between p65 NF-kB and in vitro response to fludarabine. Sensitivity to fludarabine was evaluated using LD50 values calculated from sigmoidal dose-response curves. These LD50 values positively correlated with nuclear p65 NF-kB activity (r2 = 0.25; P = 0.006) indicating that in vitro drug resistance to fludarabine may be mediated, at least in part, through the transcriptional effects of NF-kB. Taken together, our data indicate that NF-kB is a key regulator of both spontaneous and drug induced apoptosis in CLL cells and is therefore a promising therapeutic target for the treatment of this incurable condition.
Individualized Dosing of Fludarabine (FLU) for Low Grade Lymphoproliferative Disorders Results in Administration of Less Medication with Fewer Infections and Equivalent Survival.
Authors: Pasquale Donald , Ramanathan Nalini , Scarpace Sarah L. , Nguyen Hung , Jackson Amy I. , Ramanarayanan Jeyanthi 
Published: April 2020

Abstract
When used in the treatment of low-grade lymphoproliferative disorders, common practice is to administer 6 monthly cycles of FLU 25-mg/m2 daily × 5. This practice is reported to cause infections (including opportunistic such as PCP and Listeria) in over 50% of patients. It has been our practice to individualize the number of monthly cycles of FLU per treatment course based on response in general administering 1 cycle of therapy when rapid and substantial response is obtained, or 1 cycle past best response. Our objective is to describe our experience with this dosing schema - incidence of infections in relation to the number of cycles of FLU administered per treatment course and overall survival. We conducted a retrospective chart review of medical records of all patients who received FLU at the Stratton VA Medical Center. We defined one treatment course as the number of five day monthly FLU doses received. Any time interval of 3 or more months between FLU was considered to indicate a new treatment course. All infectious episodes of NCI grades 3 or higher that occurred within 6 months of the last FLU treatment course were attributed to that prior treatment course. Survival was estimated by Kaplan-Meier analysis from date of diagnosis to death or last known alive. There were 15 patients treated for CLL, 9 for low-grade lymphoma, and 4 for macroglobulinemia. Three individuals treated with FLAG for ANLL were excluded. These 28 patients received a total of 44 treatment courses (each treatment course with 1 to 6 monthly cycles) of FLU. Fifty-four (54) percent of the treatment courses were with ≤2 cycles. Mean ± SD age of the patients was 65 ± 11 years.
Number of FLU cycles per treatment course Number of treatment courses with infection Number of treatment courses without infection % treatment courses with infection 1 3 12 20 2 1 8 11 3 6 3 67 4 3 1 75 5 3 1 75 6 2 1 67
The overall incidence of NCI grade 3 or higher infections was significantly increased with increased numbers of FLU cycles per treatment course (complex Chi-square = 0.0193). Treatment cycles with ≤2 compared with 3 or more cycles per treatment course (Chi-square with Yate’s correction = 0.0008) had significantly fewer infections. Median overall survival was 158 months with 78 ± 44 months of follow-up. Our data indicates that individualized dosing of FLU based on response results in using lower numbers of monthly cycles per treatment course, lower incidence of severe infections, and equivalent (based on historical controls) overall survival.
A Multi-Center, Open-Label Study To Evaluate the Safety and Efficacy of Pentostatin, Cytoxan, and Rituxan (PCR) in the Treatment of Previously Untreated or Treated, Stage II, III or IV CLL.
Authors: Mena Raul R. , Robles Robert , Auerbach Michael , Moezi Mehdi M. , Headley David 
Published: April 2020

Abstract
The decision to treat indolent CLL is often based on progressive bulky disease, worsening symptoms, and increasing hematological derangement. When treatment is indicated, these lymphoproliferative disorders are sensitive to combination immunochemotherapies utilizing pentostatin (P), a purine analog, cyclophosphamide (C), a DNA alkylator, and rituximab (R), an anti-CD20 monoclonal antibody. P+C+R represents a promising new approach in the treatment of patients with CLL. Eligibility criteria allow both treated and treatment-naïve patients diagnosed with Stage II/III/IV CLL (modified Rai classification) to be enrolled. Treatment consisted of intravenous infusions of P (4 mg/m2), C (600 mg/m2), and R (375 mg/m2) on day 1 of a 21-day cycle for a total of up to 10 cycles. Clinical evaluation (including CT scan) was performed after cycles 2, 4, 6, 8 and 10 (if necessary). The intent-to-treat (ITT) population consisted of 70 CLL patients (median age 64, range 35–83) who received a total of 286 cycles (median 4.5 per patient). ECOG status at enrollment was 0 (68%), 1 (28%) and 2 (4%). The overall response rate (CR+Cru+PR) of the 50 evaluable patients was 52% (CR 8%, Cru 14%, PR 30%, SD 46% and PD or RD 2%). 2 grade 4 neutropenia and 1 grade 4 respiratory distress was documented. 4 deaths have been recorded, all in elderly patients (&gt;80 years old), due to acute respiratory failure, myocardial infarction, pulmonary edema and sepsis. This immunochemotherapeutic regimen is active in Stage II/III/IV CLL and the incidence of significant toxicities was low with deaths occurring only in elderly (&gt;80 years old) patients. The study results will be updated at the meeting.