bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Incidence of Leukemia and Mortality from Sepsis in Patients with Severe Congenital Neutropenia Receiving Long-Term G-CSF Therapy.
Authors: Rosenberg Philip S. , Alter Blanche P. , Bolyard Audrey A. , Bonilla Mary A. , Boxer Laurence A. , Cham Bonnie , Fier Carol , Freedman Melvin , Kannourakis George , Kinsey Sally , Schwinzer Beate , Zeidler Connie , Welte Karl , Dale David C. 
Published: April 2020

Abstract
Background: In patients with severe congenital neutropenia (SCN), mortality from sepsis is reduced by treatment with granulocyte colony-stimulating factor (G-CSF), but myelodsyplastic syndrome and acute myeloid leukemia (MDS/AML) have been reported in treated and untreated patients.
Methods: We studied 374 patients with SCN and 29 patients with Shwachman-Diamond Syndrome (SDS) on long-term G-CSF enrolled in the Severe Chronic Neutropenia International Registry.
Results: In SCN, mortality from sepsis was stable at 0.9%/year. The hazard of MDS/AML increased significantly over time, from 2.9%/year after 6 years to 8.0%/year after 12 years on G-CSF. After 10 years, the cumulative incidence was 8% for mortality from sepsis and 21% for MDS/AML. The hazard of MDS/AML increased with the dose of G-CSF. Twenty-nine percent of SCN patients received more than the median dose of G-CSF (≥8 μg/kg/day), but achieved less than the median absolute neutrophil count (ANC) response (ANC &lt;2188 cells/μL at 6–18 months). In these less responsive patients, the cumulative incidence of adverse events was highest: after 10 years, 40% developed MDS/AML and 14% died of sepsis, compared to 11% and 4%, respectively, of more responsive patients whose ANC was above the median on doses of G-CSF below the median. An intermediate group achieved an ANC above the median on doses of G-CSF above the median; among them, the 10 year cumulative incidence was 15% for MDS/AML and 3% for mortality from sepsis. In secondary analyses, we found that pre-treatment blood cell counts could not predict the subsequent clinical outcome. Furthermore, on therapy, patients who were less responsive vis-à-vis their neutrophil counts had similar increases in eosinophils, basophils, monocytes, and lymphocytes, and similar decreases in platelets, as other patients maintained on ≥8 μg/kg/day. Consistent with the SCN results, in SDS patients, the limited available data do not suggest that G-CSF therapy is a risk factor for MDS/AML in SDS.
Conclusions: The risk of MDS/AML was similarly low in all patients who achieved an ANC above the median on any dose of G-CSF. It appears that G-CSF has reduced mortality from sepsis, and revealed the underlying leukemic predisposition of SCN.
Gene Expression Profiling Identifies Distinct Subclasses in Core Binding Factor Acute Myeloid Leukemia.
Authors: Bullinger Lars , Kurz Stephan , Dohner Konstanze , Scholl Claudia , Frohling Stefan , Schlenk Richard F. , Dohner Hartmut , Pollack Jonathan R. 
Published: April 2020

Abstract
Recurrent cytogenetic aberrations have been shown to constitute markers of diagnostic and prognostic value in acute myeloid leukemia (AML). However, even within well-defined cytogenetic AML subgroups with an inv(16) or a t(8;21) we see substantial biological and clinical heterogeneity which is not fully reflected by the current classification system. Therefore, we profiled gene expression in a large series of adult AML patients with core binding factor (CBF) leukemia [inv(16) n=55, t(8;21) n=38] using a whole genome DNA microarray platform in order to better characterize this disease on the molecular level. By unsupervised hierarchical clustering based on 8556 filtered genes we observed that our CBF leukemia samples separated primarily into three different subgroups. While two of the subgroups were characterized by inv(16) and t(8;21) cases, respectively, the third subgroup contained a mixture of both cytogenetic groups. There was no obvious correlation with known secondary aberrations or molecular marker like FLT3-ITD, NRAS and KIT mutations between the cases in the mixed subgroup and the others. However, the newly defined inv(16)/t(8;21)-subgroup (n=35) was characterized by distinct clinical behavior with shorter overall survival times (P=0.029; log rank test) compared to the other two groups. Unsupervised analyses within the inv(16) and t(8;21) cases also revealed corresponding inv(16) and t(8;21) subgroups with a strong trend towards inferior outcome (P=0.11 and P=0.09, respectively; log rank test). Since the primary translocation/inversion events themselves are not sufficient for leukemogenesis, distinct patterns of gene expression found within each of these cytogenetic groups may suggest alternative cooperating mutations and deregulated pathways leading to transformation. Therefore, we performed a supervised analysis to determine the characteristic gene expression patterns underlying the cluster-defined subgroups. We identified 528 genes significantly differentially expressed between the newly defined inv(16)/t(8;21)-subgroup and the other CBF cases (significance analysis of microarrays, false discovery rate &lt; 0.001). Potential candidates for cooperating pathways characterizing the mixed inv(16)/t(8;21)-subgroup included e.g. AVO3, a member of the mTOR pathway, oncogene homologs like LYN and BRAF, as well as FOXO1A and IL6ST which have been previously identified to correlate with outcome in AML (Bullinger et al., N Engl J Med350:1605, 2004). In conclusion, while the observed signatures remain to be validated for their functional relevance, both supervised and unsupervised methods provide numerous insights into the pathogenesis of CBF AML, identifying clinically significant patterns of gene expression, as well as candidate target genes involved in leukemogenesis.
The Impact of Histologic Grade on the Outcome of High-Dose Therapy and Autologous Stem Cell Transplantation for Follicular Lymphoma.
Authors: Pham Rosalyn N. , Gooley Ted A. , Keeney Grant E. , Press Oliver W. , Pagel John M. , Petersdorf Stephen H. , Maloney David G. , Gopal Ajay K. 
Published: April 2020

Abstract
Follicular lymphomas (FL) represent approximately one-fourth of all non-Hodgkins lymphomas (NHL) and are the most common indolent lymphomas. Studies suggest that patients (pts) with relapsed FL treated with high-dose therapy (HDT) and autologous stem cell transplantation (ASCT) have prolonged progression-free survival (PFS) and overall survival (OS) as compared to those treated with standard chemotherapy alone, yet the impact of histologic grade on these outcomes has not been established. In order to address this issue, we evaluated all FL pts that underwent HDT and ASCT at our center from Dec 1985 to June 2005 (n=219). Pts with transformation to diffuse large B-cell lymphoma were excluded. Histologic grades 1, 2, and 3 were present in 106 (48%), 75 (34%), and 38 (17%) pts, respectively. Other baseline characteristics at the time of transplant for this cohort included: Female = 39%, median age = 48 (range 24 – 66), stage III/IV = 93%, elevated LDH = 27%, median number of prior regimens = 3 (range 1 to 11), prior radiation therapy (RT) = 16%, bulk ≥5cm = 19%, chemosensitive = 68%, complete remission = 18%. 53% of pts are alive and 36% are alive without relapse at last contact leading to 5- and 10-year estimates of 60% and 42% for OS and 39% and 28% for PFS, respectively. The median follow-up for surviving patients was 7.8 years. The 5- and 10-year OS estimates for the histologic grades were grade 1: 57% and 42%, 2: 60% and 36%, and 3: 65% and 55% (Figure). Five- and 10-year PFS estimates were grade 1: 35% and 27%, grade 2: 44% and 25%, and grade 3: 45% and 40% (Figure). In order to adjust for any potential imbalances of critical prognostic factors between the three grades, we performed a multivariable analysis adjusting for criteria that were found to be associated with OS and/or PFS (age, # of prior regimens, prior RT, chemosensitivity, and LDH) and obtained the hazard ratios (HR) and p-values noted (Table). These data suggest that 1) prolonged OS and PFS can be attained via HDT and ASCT in FL pts, 2) histologic grade does not statistically significantly impact outcomes, and 3) other factors noted above should be utilized to predict outcome and counsel patients.
Grade HR for Death (95% CI) p HR for Death or Progression (95% CI) p 1 1 (– –) – 1 (– –) – 2 1.00 (0.66–1.54) 0.98 0.92 (0.63–1.34) 0.69 3 0.66 (0.35–1.24) 0.20 0.79 (0.48–1.33) 0.38 Figure Figure
Crystal Structure of the Interaction of Human Urokinase Plasminogen Activitor with Its Receptor.
Authors: Huai Qing , Mazar Andrew P. , Parry Graham , Kuo Alice , Cines Douglas B. , Li Yongdong , Yuan Cai , Bian Chuanbing , Chen Liqing , Furie Bruce , Furie Barbara C. , Huang Mingdong 
Published: April 2020

Abstract
Urokinase-type plasminogen activator (uPA) and its cellular receptor (uPAR) mediate plasminogen activation. The uPA binds to uPAR with high affinity (Kd 0.1–1nM), thus localizing the generation of plasmin from plasminogen on the surface of a variety of cells. uPA-uPAR binding is also involved in other cellular functions and diverse pathophysiological processes such as tissue remodeling during wound healing, atherosclerosis, angiogenesis and tumor metastasis. We have determined the structure of uPAR complexed with the amino terminal fragment (amino acid residues 1–143) of uPA (ATF), which contains the uPAR binding domain, at 1.9 Å resolution by X-ray crystallography. Soluble uPAR (suPAR) and the ATF were expressed separately in stably transfected Drosophila Schneider 2 (S2) cells. The suPAR-ATF complex was crystallized by the sitting-drop vapor diffusion method. However, the diffraction of this crystal was limited to 3.1 Å resolution. Therefore, the Fab fragment of an antibody raised against suPAR, ATN-615, was used to facilitate suPAR-ATF crystallization. Crystals of the suPAR-ATF-ATN615 ternary complex were generated by microdialysis with 4% PEG4K, 5% ethylene glycol, 5% methanol, 0.05% sodium azide, 50 mM cacodylate pH 6.5. A complete data set of the ternary complex to 1.9 Å was collected using synchrotron radiation at the Advanced Photon Source (APS), Argonne National Laboratory. The crystals belong to the monoclinic space group, with unit cell parameters a=51.79 Å, b=86.81 Å, c=124.69 Å and β=94.54°. uPAR is comprised of three consecutive domains (D1, D2 and D3) that form the shape of a thick-walled teacup with a diameter of about 52 Å and a height of 27 Å. At the center of teacup and surrounded by three suPAR domains is a cone shape cavity with a wide opening (25 Å) and large depth (14 Å). ATF consists of a growth factor domain (GFD) and a kringle domain. Both domains pack more tightly in the complex structure compared with their unbound state. The GFD domain of uPA occupies part of the uPAR cavity and is primarily responsible for uPAR binding. The D1 domain of uPAR forms three hydrogen bonds and many hydrophobic interactions with the GFD domain of uPA, thus playing an important role in the binding of uPA. However, D2 and D3 of uPAR also have direct interactions with the GFD domain of uPA. The kringle domain of uPA sits outside the uPAR pocket, but forms some direct contacts with the D1 domain of uPAR. Therefore, the three domains of uPAR and two domains of uPA form a complementary interaction, which describes the structural basis for the high affinity binding of uPA to uPAR. This structure presents the first high resolution view of uPA-uPAR interaction, and may provide a new platform to design de novo uPA-uPAR antagonists.
Tissue Factor Activation Involves Disulfide Switching.
Authors: Chen Vivien M. , Ruf Wolfram , Berndt Michael C. , Hogg Philip J. 
Published: April 2020

Abstract
Tissue factor (TF), the essential cofactor for FVIIa, is required for activation of FX and FIX to generate thrombin. Transmembrane TF resides in a cryptic configuration on the cell surface with low procoagulant activity, however TF can be rapidly switched to an active configuration on exposure to ionomycin. The precise nature of this switch is unknown, however it is known to be an extracellular event (Wolberg et al Biophys Biochem Comm.272, 2000). The extracellular part of TF consists of 2 fibronectin type III domains. The disulphide-bond in the membrane proximal domain (Cys186–Cys209) is atypical for fibronectin domains in that it links adjacent strands in the same β-sheet, a cross-strand bond. The Cys186-Cys209 TF bond has the same unusual configuration as the disulphide-bond in the second domain of CD4, which controls CD4 function by switching between oxidized (disulphide) and reduced (dithiol) states (Matthias et al. Nature Immunol.3, 727, 2002). Ablation of the cross-strand bond severely impairs procoagulant activity (Rehemtulla et al. J. Biol. Chem.266, 10294, 1991). Labeling with a biotinylated maleimide, we demonstrate that cryptic tissue factor is reduced at the domain 2 disulfide and oxidised on activation. In HL60 cells, membrane based tissue factor procoagulant activity is blocked by the mono-thiol alkylator N-ethylmaleimide but increased by formation of the disulfide via the thiol oxidiser, HgCl2 or thiol cross-linkers, bismaleimidohexane and bismalemidoethane. These results indicate that the activation of TF involves a change of conformation of the domain 2 of TF caused by formation of the cross-strand cys186–cys209 disulfide bond. This is likely to be the physiological change that facilitates productive binding of FIX and FX in coagulation.
Rituximab Plus GM-CSF for Patients with Chronic Lymphocytic Leukemia.
Authors: Ferrajoli Alessandra , O’Brien Susan M. , Faderl Stefan H. , Wierda William G. , Ravandi-Kashani Farhad , O’Neal Brandi , Schlette Ellen , Albitar Maher , Kantarjian Hagop M. , Keating Michael J. 
Published: April 2020

Abstract
Treatment with single agent rituximab is associated with overall response (OR) rate of 15–35% in previously treated patients and up to 44% in untreated patients with CLL. The majority of the responses are partial (PR). GM-CSF was found to increase surface CD20 expression in vitro and potentiate ADCC and therefore improve the efficacy of rituximab. The combination of rituximab and GM-CSF has been investigated in follicular lymphomas and found to be associated with significant responses (McLaughlin et al. Ann. Oncol16:v68, 2005). Therefore, we designed a phase II study of rituximab plus GM-CSF in CLL. Patients were eligible for this study if they belonged to one of the following: Group 1: previously untreated patients, Rai stage 0-II and b2M &gt;3 mg/mL, B symptoms or significant fatigue. Group 2: previously untreated patients ≥ 70 years with indication for treatment (NCI-WG criteria). Group 3: previously treated patients with evidence of active disease (NCI-WG criteria). All patients received 4 weekly infusions of rituximab 375 mg/m2 plus GM-CSF 250 mcg sc three times a week for 8 weeks. Prior to study entry patients were evaluated for genomic aberration (FISH), IgVH mutation status, ZAP70 expression, and number of CD20 antigen sites. Changes in self-reported fatigue were measured using the FACT-An fatigue scale. Eighty-two patients have been enrolled at this time and 55 are evaluable for response and toxicity. Responses (NCI-WG):
Pts CR (%) NPR (%)* PR (%) OR (%) *Nodular PR:residual lymphoid nodules present Group 1 9 7 7 (78) Group 2 20 3 2 12 17 (85) Group 3 26 2 3 9 14 (54) OVERALL 55 5 (9) 5 (9) 28 (51) 38 (69)
Toxicities attributable to the administration of GM-CSF consisted in Grade 1 injection site reactions in 25% and Grade 1–2 bone pain in 13%. Grade 3 thromobocytopenia was observed in 2% of the patients and Grade 3–4 neutropenia in 4%. Grade 3 infections were observed in 9% of the patients.
The following characteristics were correlated with responses.
Characteristic Patients CR% OR% FISH 11q-/17p-11 1 (9) 4 (36) 13q-/+12/normal 38 4 (11) 32 (84) p&lt;.001 IgVH Unmutated 23 1 (4) 11 (48) Mutated 28 4 (14) 23 (82) p=.01 ZAP70 Positive 30 1 (3) 17 (57) Negative 23 4 (17) 17 (74) p=NS CD20 sites/cell ≤10,000 20 10 (50) &gt;10,000 16 3 (19) 13 (81) p=.05
Treatment with rituximab plus GM-CSF was associated with a reduction of self-reported fatigue (median score reduction 7 points) in two thirds of the patients.
In conclusion, the combination of rituximab plus GM-CSF is well tolerated and associated with an encouragingly high response rate. Favorable genomic profile, mutated IgVH status and high level of CD20 expression predicted for response to this combination. Ongoing correlative studies aim to clarify the mechanism of action of this combination. Accrual to this study is ongoing.
The Interaction of the Wnt and Notch Pathways Modulates NK vs. T Cell Commitment.
Authors: Aoyama Keisuke , Varnum-Finney Barbara , Moon Randall T. , Bernstein Irwin D. 
Published: April 2020

Abstract
The Wnt and Notch signaling pathways have critical roles in cell fate decisions. However, the interaction of these pathways is poorly understood. Using highly purified Wnt3a and immobilized Notch ligand, Delta1ext-IgG, we investigated the mechanisms involved in Wnt and Notch signaling interactions during hematopoietic stem cell differentiation. When CD34+CD38- cord blood stem cells were cultured for 2 to 3 weeks with five growth factors (SCF 300ng/ml, Flt-3L 300ng/ml, TPO 100ng/ml, IL-6 100ng/ml, and IL-3 10ng/ml), most precursor cells lost CD34 expression and differentiated into mature cells, most of which were monocytes. However, as we previously reported, when cells were cultured with Delta1ext-IgG, we found an increased percentage of lymphoid progenitors (CD34+CD7+) and more mature lymphoid precursors (CD34-CD7+) in the cell population. The addition of purified Wnt3a (100ng/ml) alone without Delta1ext-IgG did not significantly change the percentage of CD7+ cells (1% vs. 3%). However, when both Wnt3a and Delta1ext-IgG were added, we saw an increased percentage of CD7+ cells (11% with Delta1ext-IgG alone to 27% with both) (Fig. 1A). Wnt3a also enhanced gene expression of CD3ε and preTα induced by Delta1ext-IgG. These results suggest that Wnt3a enhances the effect of Notch signaling on T-cell lineage development. To test the role of endogenous Wnt signaling, we added Dickkopf1, an inhibitor of Wnt signaling. When CD34+CD38- cells were cultured with Dickkopf1 (300ng/ml) alone for a 2 week, the percentage and number of CD56+ NK cells was unaffected. However, when cultured with Dickkopf1 and Delta1ext-IgG, the percentage and number of CD56+ NK precursor cells were increased (2% with Delta1ext-IgG alone vs. 17% with Dickkopf1 and Delta1ext-IgG; 0.2 ×105 vs. 1.5×105, p&lt;0.01) (Fig. 1B). This result shows that decreased endogenous Wnt signaling enhances the generation of NK cells in the presence of Notch signaling. To address whether Wnt signaling affects Notch signaling by modulating protein levels, we assessed the amount of activated Notch1 intracellular domain with or without Wnt 3a by Western blot (Fig. 2A). After stimulation for 24 h, Wnt3a increased the amount of activated Notch1 intracellular domain induced by Delta1ext-IgG. Wnt3a also enhanced the expression of a primary target gene of Notch signaling, Hes1, determined by quantitative RT-PCR (Fig. 2B). In contrast, Dickkopf1 reduced Delta1ext-IgG-induced Hes1 expression. These results suggest that Wnt signaling directly modulates Notch signaling. Thus, these studies suggest that Wnt signaling is a key factor in cell fate determination at the point of NK/T cell commitment that is mediated via an interaction with Notch signaling. These studies also suggest that Wnt directly regulates Notch signaling by modulating protein turnover.
Fig. 1, Fig. 2 Fig. 1, Fig. 2.
Decreased Responsiveness of Mature Dendritic Cells from Cord Blood to Chemotaxis Induced by CXCR4 Ligand CXCL12, and CCR7 Ligand CCL19 May Explain Lowered GVHD Associated with Cord Blood Tranplantation.
Authors: Li Geling , Han Myung-Kwan , Broxmeyer Hal E. 
Published: April 2020

Abstract
Cord blood has been successfully used as a transplantable source of hemopoietic stem cells with a relatively low incidence of graft-versus-host disease (GVHD). Mature dendritic cells (DC), as the most efficient antigen presenting cells, play a critical role in the pathogenesis of GVHD and their function is intimately linked to their capacity to migrate. Chemokines and their G-protein linked receptors are of particular importance in DC migration. CCR7 and CXCR4 are two chemokine receptors expressed on mature DC. Binding of CCR7 ligand, MIP-3β/CCL19 and CXCR4 ligand, SDF-1/CXCL12 induce chemotaxis of mature DC and are pivotal for DC to initiate their functions in vivo. We hypothesized that the relatively lowered level of GVHD elicited by cord blood, compared to adult cells, might be due, at least in part, to a reduced capability of cord blood DC to migrate to lymphoid organs to initiate immune reactions and to organs such as liver and skin where GVHD pathogenesis usually occurs. To begin testing this hypothesis, we randomly picked 16 cord blood donors and 14 adult blood donors and did paired comparison study on chemotactic responses of cord blood and adult blood DC to CXCL12 and CCL19. Monocytes were isolated from both cord blood and adult blood and cultured with GM-CSF and IL-4 for 5 days to generate immature DC. LPS was added into cell culture for an additional 1 day to induce the maturation of these DC. Mature DC were harvested on day 6. We found that cord blood DC expressed significantly reduced surface levels of CCR7 compared with that of adult blood DC (p = .00067) and migrated at significantly lower efficiency towards the CCR7 ligand, CCL19 than did adult blood DC (p=0.029). Cord blood DC expressed significantly higher surface expression of CXCR4 than adult blood DC (p = .0018), but migrated at significantly lower efficiency towards CXCR4 ligand, CXCL12 than adult blood DC (p=0.00012). The lowered responsiveness of mature cord blood DC to chemotaxis by CXCL12 may reflect the enhanced truncation of CXCR4 we found on these cells. Also, cord blood DC migrated less well than adult blood DC toward the combination of CXCL12 and CCL19 than adult blood DC (p = .0014). These results suggest that low incidence and pathogenesis of GVHD observed in cord blood might be due, at least in part, to reduced capability of cord blood mature DC to migrate to critical organs where immune reaction and inflammation occur, effects reflecting lowered expression of CCR7 and enhanced truncation of CXCR4 on mature cord blood DC.