bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dose-Dense Rituximab Improves Outcome of Elderly Patients with Poor-Prognosis Diffuse Large B-Cell Lymphoma (DLBCL): Results of the DENSE-R-CHOP-14 Trial of the German High-Grade Non-Hodgkin Lymphoma Study Group (DSHNHL).
Authors: Pfreundschuh Michael , Zeynalova Samira , Poeschel Viola , Haenel Mathias , Schmitz Norbert , Hensel Manfred , Reiser Marcel , Loeffler Markus , Schubert Joerg 
Published: November 2019

Abstract
Background: While 6 to 8 cycles of CHOP in combination with rituximab are widely accepted as standard regimen of care for aggressive lymphomas, the optimal dose and number of rituximab application have not been determined to date. In a previous pharmacokinetic study we had shown that the concomitant application of CHOP and rituximab does not achieve a plateau of rituximab trough levels until cycle 5 (Reiser, Blood 108, 778a, 2006). In order to achieve high rituximab levels early, we increased the number of rituximab applications.
Methods: 100 elderly patients with aggressive CD20+ B-cell lymphoma received 6 cycles of biweekly CHOP-14 combined with 12x rituximab (375 mg/m2) on days 0, 1, 4, 8, 15, 22, 29, 43, 57, 71, 85, and 99. Radiotherapy was planned to sites of initial bulk and/or extranodal involvement. The primary endpoint was event-free survival (EFS). 306 patients treated within the RICOVER-60 trial (Pfreundschuh et al., Blood 64a, 2006) with 6xCHOP-14 and 8 applications of rituximab served as control.
Results: 97/100 patients are evaluable for response. Dose-dense rituximab resulted in plateau trough serum levels of rituximab as early as day 1 of the first chemotherapy cycle and higher rituximab levels were maintained throughout the treatment compared to 8 bi-weekly applications. Because 3 therapy-associated deaths were observed among the first 20 patients treated, prophylaxis with levofloxacin, acyclovir and cotrimoxazol became mandatory for the following patients. Despite a less favorable study population DENSE-R-CHOP-14 resulted in a somewhat higher complete remission (83% vs. 78%) and lower progression under therapy rate (5% vs. 7%) rate, but event free and overall survival were not different compared to 8 biweekly applications of rituximab. However, a subgroup analysis of patients according to IPI risk group showed that DENSER-R-CHOP-14 resulted in a higher complete response rate of patients with poor-prognosis (IPI:3–5) disease (81% vs. 68%). This advantage translated into a better 1-year event-free survival rate (74% vs. 65%) of these patients.
Conclusion: In combination with 6 cycles of CHOP-14 densification of rituximab achieves higher rituximab serum levels and results in higher complete remission and event-free survival rates in elderly patients with poor-prognosis DLBCL. These observations from a phase-II trial must be confirmed in a randomized study.
Development of Osteonecrosis Is Associated with Difference in Upregulation of Antithrombin, Protein C and Protein S after Four Weeks of Dexamethasone during Induction Treatment in Childhood Acute Lymphoblastic Leukemia.
Authors: te Winkel Mariel L. , Pieters Rob , Appel Inge M. , Van Den Heuvel Marry M. 
Published: November 2019

Abstract
Osteonecrosis (ON) is a serious complication of chemotherapy for pediatric acute lymphoblastic leukemia (ALL). Impairment of microcirculation seems to be involved in the pathogenesis of ON. We hypothesized that variation in hypercoagulability plays a role in the etiology of ON during treatment of ALL. Dexamethasone (dexa) and asparaginase (asp) are important drugs in the induction treatment of pediatric ALL that may influence the coagulation system. In the current study differences in coagulation parameters between pediatric ALL patients with and without ON were evaluated. Blood samples were taken at diagnosis and at several time-points during induction of the DCOG-ALL9 treatment schedule. Procoagulant factors (fibrinogen, factor II,V,VII,IX,X), anticoagulant factors (antithrombin (AT), protein C and protein S), parameters of thrombin generation (prothrombin fragment 1+2 (F1+2), thrombin antithrombin (TAT) complex) and of fibrinolysis (α2-antiplasmin (a2AP), plasminogen, plasmin-α2AP (PAP) complex, D-dimers) were determined. In total 137 patients without ON and 24 patients with ON were evaluated. In addition plasminogen activator inhibitor type 1 (PAI-1) activity and tissue plasminogen activator antigen (tPA-ag) concentration were studied in 25 patients, 15 without ON and 10 with ON. At diagnosis no significant differences in any of the coagulation parameters were found between patients with and without ON. After 4 weeks dexa treatment (day 29), AT, protein-C activity and protein-S total (anticoagulants) were significantly less upregulated in ON positive patients as compared to ON negative patients (respectively p=0.043, p=0.029, p=0.023). During tapering of dexa and administration of 4 doses asp at day 29, 33, 36 and 40 these parameters significantly decreased in both ON positive and negative patients. However, the nadir of these anticoagulant proteins reached lower levels in the ON positive patients as compared to ON negative patients and in ON positive patients levels even reached below lower limits of normal reference ranges. No other dexa or asp induced differences in any of the other coagulation parameters were found between ON negative and positive patients. These results indicate that development of ON in pediatric ALL patients seems to be associated with less upregulation of AT, protein C and protein S after 4 weeks treatment with dexa (day 29). ON positive patients subsequently declined below the lower limit of normal reference range during tapering of dexa and administration of asp in contrast to ON negative patients. This may suggest a therapy induced hypercoagulable state contributing to the development of ON.
The Role of Autologous Stem Cell Transplantation (ASCT) in Patients with Advanced Waldenström’s Macroglobulinemia.
Authors: Kyriakou Charalampia , Canals C. , Sureda A. , Taghipour G. , Gisselbrecht C. , Mazza P. , Montserrat E. , Niederwieser N. , Indrak K. , Kolbe A. , Biron P. , Levis A. , Goldstone A. , Schmitz N. 
Published: November 2019

Abstract
Waldenström’s macroglobulinaemia (WM) is a relatively rare disorder that primarily affects elderly patients. Conventional therapies for symptomatic WM result in response rates of up to 70%. However, complete responses are rare and the disease remains incurable. Due to the indolent nature of the disease and the older age of patients the role of autologous stem cell transplantation (ASCT) in the treatment of patients with WM has not been analyzed in large series. In this retrospective multicenter study we report the outcome of 201 WM patients (132 male, 69 female), who underwent ASCT between 1992 and 2005. The median age at transplant was 53 years (22–73) and the median time from diagnosis to transplant was 18 months (3–239). Forty patients (20%) were in 1st maximum response (MR1), 24 (12%) in ≥MR2, 83 (41%) in PR1, 27 (13%) in ≥PR2 and 27(14%) were primary refractory to treatment. Conditioning regimens were BEAM (44%), TBI/Cyclophosphamide or Melphalan (28%), Melphalan (14%), BuCy (2%) and others (12%). The source of stem cells was PB in 188, BM in 10, and both in 3 patients. All patients but 3 had successful engraftment. With a median follow-up of 26 months (5–163), 112 (56%) patients are alive and free of disease, 73 (36%) patients have relapsed after a median of 14 months (1–110) post ASCT. Fifty-two patients died, 36(18%) from disease progression and 16(8%) from treatment toxicity. Non- relapse mortality was 6% at 1 year. The actuarial OS was 86% at 1 year, 75% at 3 years, and 61% at 5 years. The probability of relapse was 20% at 1 year, 38% at 3 years and 55% at 5 years with an estimated PFS of 74%, 54% and 33% at 1, 3, and 5 years respectively. Multivariate analysis revealed that, chemosensitive disease at the time of ASCT was the most important factor for NRM (p<0.001), relapse rate (p<0.01), PFS (p<0.001) and OS (p<0.001). In conclusion, this study suggests that ASCT is a safe procedure in patients with WM and that a significant proportion of heavily pre-treated patients with this disorder can respond to the procedure and achieve prolonged PFS.
Effect of the Percent Decrease in Monoclonal Protein with Induction Therapy on the Outcome after Autologous Stem Cell Transplantation in Patients with IgG and IgA Multiple Myeloma.
Authors: Reece Donna E. , Trieu Young , Pokoradi Alida , Xu Wei , Fung Sharon , Chen Christine , Kukreti Vishal , Mikhael Joseph , Trudel Suzanne , Stewart A. Keith 
Published: November 2019

Abstract
One potential approach to improve the results of autologous stem cell transplantation (ASCT) in multiple myeloma (MM) involves the use of more potent induction regimens; the achievement of deeper remission pre-ASCT should translate into longer progression-free (PFS) and overall survival (OS). To evaluate this hypothesis, we assessed the influence of the percent reduction in serum monoclonal protein (M protein) pre-ASCT in patients (pts) treated with dexamethasone-based regimens (most often VAD) on the best response, PFS and OS achieved after ASCT. Between 2000–2006, 376 pts with IgG (267) or IgA (109) non-progressive MM underwent ASCT. Median age was 59 (21–73) yrs; 60% were male. The median hemoglobin at diagnosis was 103 g/L, creatinine 98 μmol/L, β2-microglobulin 241 nmol/L and albumin 35 g/L. Maintenance therapy was given to 84 (22%) after ASCT and consisted of corticosteroids in 29, thalidomide in 10, both in 41 and other regimens in 4 pts. Patients were divided into 4 groups based on the per cent reduction in M protein after induction therapy:
group A, ≥ 99%; group B, 90–98%; group C, 50–89%; group D, < 50%.
Post-ASCT responses included complete remission (CR), very good partial remission (VGPR) (≥ 90% reduction in M protein), PR (≥ 50% reduction) and stable disease (SD). Median follow-up from diagnosis is 37.4 mos and from ASCT 24.1 mos. For all pts, the median OS from diagnosis is 90.8 mos (95% CI 73.9–129.1 mos) and from ASCT 63.9 mos (95% CI 50.8–69.4 mos), while the median PFS from ASCT is 21.3 mos (95% CI 19.1–23.3 mos). Maintenance therapy had no significant effect on PFS (p=0.49) or OS (p=0.59). The post-ASCT results in evaluable patients according to the percent reduction in M protein after induction therapy are summarized below. We conclude:
High-grade remissions after dexamethasone-based induction therapy are uncommon, with only 2% achieving ≥ 99% and 13% achieving 90–98% reduction in serum M protein; post-ASCT CR and VGPR rates were higher in these 2 groups; there was no significant difference in PFS or OS-based on protein response prior to transplant; whether newer induction regimens, which incorporate novel agents and which produce more CRs and VGPRs before ASCT, will confer better PFS and OS post-ASCT will be of great interest.
Table 1 Group N CR (%)1 VGPR (%)1 PR (%)1 SD (%)1 Median PFS (mo)2 Median OS (mo)3 1p<0.001;2p=0.77;3p=0.79 A 7 50 50 Not reached Not reached B 47 19 64 17 23.3 Not reached C 232 14 17 68 1 20.7 59.1 D 87 11 7 54 28 21.1 69.4 Figure Figure Figure Figure
Cloning of AML1-LAF4 Fusion Gene in Childhood T-Cell Acute Lymphoblastic Leukemia with t(2;21)(q11;q22) by Bubble PCR Method for cDNA.
Authors: Chinen Yoshiaki , Taki Tomohiko , Nishida Kazuhiro , Shimizu Daisuke , Okuda Takashi , Yoshida Naohisa , Kobayashi Chie , Koike Kazutoshi , Tsuchida Masahiro , Hayashi Yasuhide , Taniwaki Masafumi 
Published: November 2019

Abstract
The AML1/RUNX1 gene is frequently rearranged by chromosomal translocations in acute leukemia. To date, more than 10 fusion genes involving AML1 have been cloned, such as AML1-MTG8 in acute myeloid leukemia (AML) with t(8;21), AML1-EVI1/MDS1 in therapy-related AML/myelodysplastic syndrome with t(3;21), and TEL/ETV6-AML1 in B precursor ALL with t(12;21). We analyzed a pediatric patient having T-cell acute lymphoblastic leukemia (T-ALL) with t(2;21)(q11;q22), and identified that the LAF4 gene on 2q11.2–12 was fused to the AML1 gene on 21q22 using the bubble PCR method for cDNA. The genomic breakpoints were within intron 7 of AML1 and intron 7 of LAF4 resulting in the in-frame fusion of exon 7 of AML1 and exon 8 of LAF4. LAF4 gene is a member of the AF4/FMR2 family, and was previously identified as a fusion partner of MLL in B-precursor ALL with t(2;11)(q11;q23), although AML1-LAF4 was in T-ALL. LAF4 is the first gene fused with both AML1 and MLL in acute leukemia. These findings provide new insights into the common mechanism of AML1 and MLL fusion proteins in the pathogenesis of ALL. In this study, we first applied the panhandle PCR method that is usually used for cloning the fusion partners of MLL or NUP98; however, no fusion transcripts could be obtained. Therefore, we searched for another method for cloning the fusion transcripts and successfully adapted the bubble PCR method for cloning the novel AML1-LAF4 fusion transcript. To date, bubble PCR has been performed for cloning unknown genomic fusion points but not fusion cDNAs. Using double-strand cDNA, we could apply the bubble PCR method for cloning fusion cDNA with rare non-specific products. Bubble PCR for cDNA could amplify in both 5′ to 3′ and 3′ to 5′ directions of the gene or transcript and handle any exons fused to unknown partners for amplification easily. This method will contribute to identifying numerous novel translocation partners more easily, and these findings may help to clarify the role of the fusion proteins in leukemogenesis.
Detection of Genomic Lesions in Childhood Precursor-B Cell ALL in Diagnosis and Relapse Samples Using High Resolution Genomic Profiling.
Authors: Kuiper Roland P. , van Leeuwen Frank N. , Keijzers-Vloet Suzanne T.M. , van Reijmersdal Simon V. , Hehir-Kwa Jayne Y. , van Wering Elisabeth R. , van der Velden Vincent H.J. , van Dongen Jacques J.M. , Schoenmakers Eric F.P.M. , van Kessel Ad Geurts , Hoogerbrugge Peter M. 
Published: November 2019

Abstract
Due to advances in therapeutic regimens developed during the last two decades, the majority of children with acute lymphoblastic leukemia (ALL) respond well to therapy. However, in approximately 25% of the patients relapses occur. Chomosome aneuploidies and recurrent chromosomal translocations are of considerable prognostic importance, and are routinely used in the course of clinical decision making. Current technological developments in molecular cytogenetic techniques have revealed that genetic lesions driving tumorigenesis frequently occur at the submicroscopic level and, consequently, escape standard cytogenetic observations. Therefore, we have previously performed high resolution genomic profiling of precursor-B-cell ALL samples obtained at diagnosis, using 250k NspI SNP-based oligoarrays from Affymetrix (Kuiper et al., 2007). By doing so, we detected multiple de novo genetic lesions, some of which were subtle and affected single genes. Many of these lesions involved recurrent (partially) overlapping deletions and duplications, encompassing various established leukemia-associated genes, such as ETV6, RUNX1, and MLL. Importantly, the most frequently affected genes were those controlling G1/S cell cycle progression (e.g. CDKN2A, CDKN1B, and RB1), followed by genes associated with B-cell development. The latter group included the B-lineage transcription factors PAX5, EBF, E2-2, and IKZF1 (Ikaros), as well as genes with other established roles in B-cell development, i.e., RAG1 and RAG2, FYN, PBEF1, or CBP/PAG. Here we have selected 34 additional precursor-B cell ALL cases that suffered from relapses 6 months to 9 years after diagnosis. Lesions affecting genes involved in G1/S cell cycle progression and B-cell development were observed with similar frequencies in the diagnosis and relapse samples as compared to our previous cohort of patients with unknown therapy response. However, additional (secondary) lesions were observed in the relapse samples in nearly all patients analyzed, indicating that these relapse samples are genomically distinct. In addition, several cases were encountered in which the diagnosis and relapse samples carried alternative lesions affecting the same gene(s), including CDKN2A and PAX5, suggesting that inactivation of these genes were secondary but essential events required to develop a full blown leukemia.
Evaluation of Platelet Behavior in Response to Varied Conditions in a Microfluidic Flow Cells.
Authors: Conant Carolyn G , Schwartz Michael A , Ionescu-Zanetti Cristian 
Published: November 2019

Abstract
Abstract 5075
Platelet function under shear flow can be tested using a variety of instruments ranging from cone and plate viscometers to microfluidic flow cells. While these methods offer relevant data, they generally suffer from high blood volume requirements, low throughput and complex operating protocols. We have developed a flow-based well plate microfluidic device that operates at very wide shear ranges (from 0-200dyn/cm2) and use minimal blood volumes ('1ml per experiment). Here we present data using the microfluidic device to examine platelet response over time on coated surfaces. Flow channels were coated with fibronectin, von Willebrand factor (vWF), and collagen I. Whole blood was introduced at increasing shear stress ranging from 0-200dyn/cm2. Devices were used to study dose responses to abciximab (ReoPro) on vWF and collagen I at 200 and 10dyn/cm2 respectively by examining percent platelet coverage following application of flow. Within the concentration range tested, 1.25-20 μg/ml (26.5nM-470nM), for both collagen and vWF, the IC50 fell slightly below 26.5nM. We compared the data generated in the microfluidic channels to data generated in traditional parallel plate flow chambers. While response to coated surfaces and ReoPro were predictably comparable, the time to set up the microfluidic device and the volume of blood used were significantly decreased, 6X and 10X respectively, leading to higher throughput in one experimental session and more data generated from one donor. Therefore, the use of such microfluidic devices can greatly streamline studies with multiple variables enabling many data points to be generated from single donors within a single device.

Disclosures
Conant: Fluxion : Employment. Schwartz:Fluxion : Employment, Equity Ownership. Ionescu-Zanetti:Fluxion: Employment, Equity Ownership.
STAT3-Activated GM-CSFRα Shuttles To The Nucleus and Protects CLL Cells From Apoptosis
Authors: Li Ping , Rozovski Uri , Harris David , Zhiming Lui , Ferrajoli Alessandra , Bueso-Ramos Carlos E. , Hazan-Halevy Inbal , Grugurevic Srdana , Wierda William G. , Burger Jan A. , O'Brien Susan , Faderl Stefan H. , Keating Michael J. , Estrov Zeev 
Published: November 2019

Abstract
Granulocyte-macrophage colony stimulating factor (GM-CSF) regulates the survival, proliferation, differentiation, and activation of hematopoietic cells, dendritic cells, and T cells. Upon GM-CSF binding, the α and β subunits of the GM-CSF receptor (GM-CSFR) dimerize and activate signaling. While exploring a possible role for GM-CSF in treating chronic lymphocytic leukemia (CLL), we found that GM-CSF did not enhance the phosphorylation of STAT3, Akt, or ERK, suggesting that GM-CSF does not directly affect CLL cells. Indeed, as in normal B cells, flow cytometry analysis of CLL cells did not detect GM-CSFRβ. However, unlike in normal B cells, GM-CSFRα (CD116) was present in CLL cells. Using confocal microscopy, cell fractionation studies, and GM-CSFRα antibody epitope mapping, we detected GM-CSFRα not only on the surfaces but also in the cytosol and nuclei of CLL cells, suggesting that GM-CSFRα has functions that are unrelated to receptor-ligand interaction. Because STAT3 is constitutively activated in CLL cells and because the promoter of the GM-CSFRα gene harbors putative STAT3 binding sites, we hypothesized that STAT3 activates GM-CSFRα in CLL cells. Both chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assay (EMSA) analyses revealed that STAT3 binds to the promoter of GM-CSFRα. We therefore cloned the GM-CSFRα promoter in multiple myeloma cell line MM1 and used luciferase assay we identified STAT3 binding activity. To investigate the function of GM-CSFRα, we induced the overexpression of GM-CSFRα in 293FT cells and analyzed the RNA and protein that co-immunoprecipitated with GM-CSFRα. We identified 7200 RNA transcripts that co-immunoprecipitated with GM-CSFRα, including SKI and MAFA oncogenes, two serine/threonine kinase genes, and DEFT1P2 and DEFP1P, which encode proteins that belong to members of the death effector domains family, known to have a role in modulating apoptosis. Overall, the transcripts that co-immunoprecipitated with GM-CSFRα belong to survival pathways, the JAK/STAT pathway, and hematopoietic lineage pathways. Mass spectrometry analysis revealed that housekeeping proteins, chaperon proteins, KAP1, and ISG-15 co-immunoprecipitated with GM-CSFRα. We found that GM-CSFRα–bound KAP1 enhanced the transcriptional activity of STAT3, whereas GM-CSFRα–bound ISG-15 inhibited the nuclear factor-κB pathway. Nevertheless, overexpression of GM-CSFRα protected MM1 cells from dexamethasone-induced apoptosis, and GM-CSFRα-siRNA induced the apoptosis of CLL cells. Taken together, our data suggest that STAT3 induces the transcription of GM-CSFRα, which has distinct, ligand-independent, anti-apoptotic activities.

Disclosures:
No relevant conflicts of interest to declare.