bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gene Expression Profiles as Prognostic Factors in Patients with Multiple Myeloma Treated with Conventional Versus Novel Agents in Correlation with Outcome.
Authors: Broyl Annemiek , Hose Dirk , de Knegt Yvonne , Lokhorst Henk , Goldschmidt Hartmut , van Duin Mark , Sonneveld Pieter 
Published: October 2019

Abstract
Background: Bortezomib (Bor), a proteasome inhibitor, and Thalidomide (Thal), an anti-angiogenic and immunomodulatory drug, have a remarkable effect in patients with relapsed or refractory MM with 30–40% response rates. In newly diagnosed patients the response rates vary from 70–85%. However, 15–30% of newly diagnosed patients do not respond to Bor or Thal. Secondly, 30% of the patients treated with these novel agents have to stop prematurely because of intolerable side effects, such as polyneuropathy, thrombocytopenia, thrombosis and gastro-intestinal symptoms.
Aim: To gain new insights regarding the mechanisms of drug response and toxicity associated with these agents, we have embarked on a prospective study to analyze gene expression profiles (GEP) of myeloma specific genes in plasma cells purified from bone marrow from myeloma patients at diagnosis who have been treated with these novel agents in order to learn which genes govern outcome upon treatment with Bor and Thal.
Methods: GEP of CD138 magnetic cell selected (MACS) myeloma plasma cells was performed using Affymetrix GeneChip Human Genome U133 plus 2.0 arrays. This program has been initiated in a large multicenter, prospective, randomized phase III trial, comparing Bor in combination with Adriamycin, Dex (PAD, arm A) followed by high dose therapy with stem cell rescue and maintenance therapy with Bor vs. Vincristine, Adriamycin and Dex (VAD, arm B) followed by high dose therapy with stem cell rescue and maintenance therapy with Thalidomide (HOVON65/GMMG-HD4). This cooperative trial in the Netherlands and Germany has started in April 2005 and has included 600 patients. Gene arrays were analyzed using correlation analysis in Omniviz. Differentially expressed genes in the different groups and in Bor responders were determined using a t-test with adjusted p-value (p<0.0001) and a 1000 permutation analysis (BRB tool).
Results: Based on correlated GEPs we divided myeloma patients into 10 clusters. We determined differentially expressed genes in the different clusters and correlated the clusters with chromosomal aberrations like 13q loss, 9 and 11 gain and translocations. Three clusters showed high expression of MMSET/FGFR3, MAF downstream targets and CCND1, respectively.
Conclusion: unsupervised cluster analysis led to the subdivision of myeloma patients into clusters, each with a specific gene expression signature. We determined differentially expressed genes in the different clusters and could confirm 3 subgroups (MMSET/FGFR3, MAF/MAFB, and a group with high CCND1 expression) as have been published before.
Laboratory Response to Intranasal Desmopressin in Women with Menorrhagia and Underlying Platelet Dysfunction.
Authors: Rose Shelonitda S. , Faiz Ambarina , Miller Connie H. , Saidi Parvin , Philipp Claire S. 
Published: October 2019

Abstract
Intranasal desmopressin (IN-DDAVP) is used for home treatment of menorrhagia in women with inherited bleeding disorders. The effect of IN-DDAVP on laboratory hemostatic parameters in women with menorrhagia and associated platelet dysfunction is unknown. We evaluated the effects of IN-DDAVP on hemostatic parameters in women with menorrhagia and platelet dysfunction and correlated them with menstrual blood flow. Eleven women (aged 18–45) with menorrhagia and abnormal platelet aggregation and/or platelet adenosine tri-phosphate (ATP) release had determination of factor VIII coagulant activity (FVIII: C), von Willebrand factor antigen (VWF: Ag), von Willebrand factor ristocetin cofactor (VWF: RCo) activity, platelet aggregation, and platelet ATP release pre and 60 minutes post IN-DDAVP. Eight of eleven women underwent Platelet Function Analyzer (PFA-100) closure time determination with collagen/epinephrine (CEPI) and collagen/adenosine diphosphate (CADP) cartridge pre and post treatment. IN-DDAVP was administered during two consecutive menstrual cycles. Menstrual blood flow was assessed during each cycle using a pictorial blood assessment chart (PBAC). Following administration of IN-DDAVP there was a 2–3 fold increase in FVIII: C, VWF: Ag, and VWF: RCo observed in most patients over baseline levels. Mean FVIII:C increased from 130% ± 34.8% to 218% ± 96.5% (P < 0.007), VWF: Ag increased from 86.9% ± 35.1% to 128.7% ± 64.3% (P < 0.011), and VWF: RCo increased from 89.8% ± 42.8% to 139.6% ± 86.5% (P < 0.02). The median baseline PBAC score of 235 decreased to 136 following IN-DDAVP. In addition, there were significant inverse correlations between changes in PBAC score and changes in VWF: Ag (rs = − 0.85, p = 0.02), VWF: RCo (rs = −0.81, p = 0.029), and FVIII:C (rs = 0.92, p = 0.003) with IN-DDAVP. Mean PFA-100 closure time with the CEPI cartridge shortened from 166.8sec ± 85sec to 90.9sec ± 49.3sec (p< 0.02), and the closure time with the CADP cartridge shortened from 104.5sec ± 34.3sec to 64.8sec ± 40.4sec (p <0.007). There were significant inverse correlations between post IN-DDAVP PFA-100 CADP closure time and post IN-DDAVP FVIII:C (rs = −0.86, p = 0.005), VWF: Ag (rs = −0.72, p = 0.04), and VWF: RCo (rs = −0.80, p = 0.01). In addition there were also significant inverse correlations between post IN-DDAVP PFA-100 CEPI closure time and post IN-DDAVP FVIII:C (rs = −0.73, p = 0.03), and VWF: RCo (rs = −0.74, p = 0.03), but not VWF: Ag (rs = −0.66, p = 0.07). In-vitro platelet aggregation and platelet ATP release response did not correct and did not correlate with changes in menstrual blood flow. Our results demonstrate a correlation between VWF parameters and menstrual blood flow following IN-DDAVP in women with menorrhagia and underlying platelet dysfunction.
Proteins of the Calnexin Cycle Regulate αIIbβ3 Surface Expression in Human Megakaryocyte-Lineage Cells.
Authors: Yazdani-Abyaneh Mahmoud , Mitchell W. Beau 
Published: October 2019

Abstract
The αIIbβ3 integrin receptor mediates platelet aggregation, and individuals without functional αIIbβ3 manifest the mucocutaneous bleeding disorder Glanzmann thrombasthenia. Since platelets also participate in the pathophysiology of thrombosis, anti-αIIbβ3 agents have been successful in reducing mortality after percutaneous coronary interventions and from platelet-mediated coronary thrombosis. Accordingly, much research has focused on the structure, activation and signaling of the mature receptor on the cell surface. There is much to be gained medically and scientifically by understanding the intracellular processes by which integrins are assembled and expressed. Some forms of Glanzmann thrombasthenia result from mutations that disrupt normal integrin folding and assembly, causing the integrin subunits to be retained in the ER and degraded. These cases indicate the existence of a quality control mechanism for αIIbβ3 by which misfolded subunits are recognized, retained and degraded. Previous studies in HEK293 cells and murine megakaryocytes have identified the calnexin cycle as part of this quality control mechanism. We have expanded upon these findings and studied their functional implications by determining the effect of siRNA-induced knock down of proteins from six families of post-translational processing proteins on αIIbβ3 surface expression on human megakaryocyte-lineage cells derived from umbilical cord blood (UCB): lectins, glucosyltransferases, mannosidases, HSP90s, HSP70s, BiP, protein disulfide isomerases, and tetraspanins. We report that siRNA against two proteins in the calnexin cycle decreased surface expression of natively-folded αIIbβ3 on UCB-derived megakaryocytes as measured by binding of 10E5, an αIIbβ3 complex-dependent mAb. siRNA against UDP-glucose ceramide glucosyltransferase-like 1(UGGT, a glucosyltransferase) decreased binding by 36 +/− 9%, and siRNA against EDEM1 (a mannosidase) decreased binding by 15 +/− 14%, both p < 0.05. Additionally, siRNA against CD9 increased binding by 17 +/− 8%, p < 0.05, suggesting that CD9 may be a negative regulator of αIIbβ3 surface expression. UGGT is a glucosyltransferase protein and folding sensor of the calnexin cycle that recognizes and binds to glycoproteins that are in nearly-native folding conformations, but does not bind to fully folded glycoproteins. Glycoproteins that are glucosylated by UGGT may be bound by calnexin and retained in the ER, increasing their time for folding. Fully folded proteins cannot be reglucosylated by UGGT and escape the calnexin cycle, exiting the ER. EDEM1 is a mannosidase that interacts with calnexin to accept terminally misfolded proteins and direct them to degradation. Together, these data suggest a model in which the level of expression of native-folded αIIbβ3 on the cell surface is regulated in the ER by the calnexin cycle.
Cord Blood NK Cytotoxic Activity Can Be Selelctively Modulated by Accessory.
Authors: Xing Dongxia , Li Sufang , Simon Robinson N. , William Decker , Hong Yang , David Steiner , Michael Thomas , Champlin Richard E. , McMannis John D. , Vina Marcelo F. , Cooper Laurence J.N. , Simmons Paul J. , Zweidler-Mckay Patrick A. , Shpall Elizabeth J. 
Published: October 2019

Abstract
Introduction The expansion of cord blood (CB) derived natural killer (NK) cells is a promising cell therapy approach. CB NK cell expansion and activation has proven to be feasible and effective. However, the selectivity of NK killing in an allograft setting raises concern. Accessory cells, in particular dendritic cells and mesenchymal stem cells (MSCs), have been shown to interact with and modulate NK cell proliferation and function. We explored the ability of accessory cells to modulate NK activity in the setting of clinically-relevant CB NK expansion protocols.
Methods Human MSCs were isolated from normal bone marrow donors. 5 × 106 CD56-selected NK cells or 5 × 107 unselected or CD3-depleted CB mononuclear cells (MNC) (averaging 10% CD56+ NK cells) were layered on top of the MSCs in the presence of 200u/mL IL-2. CB NK cells were tested for cytolytic activity by 51Cr release assay using MSCs and K562 AML cells. IFN-g secretion was detected by Elispot assay.
Results Cord blood-derived NK cells activated by exposure to IL-2 efficiently kill allogeneic MSCs in vitro. A 4 hour chromium release assay revealed that CD56-selected CB NK cells lysed 60% of allogeneic MSCs at E:T ratio 10:1. MSC co-culture with CB NK induced IFN-g secretion as determined by IFN-γ Elispot assay. In contrast, unselected or CD3-depleted CB MNCs with equal numbers of CD56+ NK cells were unable to lyse MSCs despite IL-2 activation. This suggests that CB accessory cells can modulate the activation or cytolytic activity of CB-derived NK cells. To address these two possibilities, we tested the ability of these CB cultures to kill a classic NK target, K562 human myeloid leukemia cells. Surprisingly, in all three CB cultures NK efficiently lysed K562 cells, 95% of lysis at 10:1 E:T ratio by chromium release assay suggesting that these accessory cells did not prevent IL-2 activation of NK cells, but selectively-modulated the cytolytic activity. Thus, preserving anti-leukemia killing while protecting non-malignant MSCs.
Conclusions Our data provide evidence for selective modulation of cord blood-derived NK cell cytolytic activity by CB accessory cells. Indeed, activation of cord-derived NK cells in the presence on CD3-depleted accessory cells yields NK cells capable of killing K562 target cells but not allogeneic MSCs. Although the underlying mechanism is unknown, this selective modulation of NK cells may be exploited in the clinical expansion of cord blood NK cells.
Severe Donor Events after Allogeneic Hematopoietic Stem Cell Donation.
Authors: Halter Jörg , Kodera Yoshihisa , Ispizua Alvaro Urbano , Greinix Hildegard , Schmitz Norbert , Favre Geneviève , Baldomero Helen , Niederwieser Diegter , Apperley Jane , Gratwohl Alois 
Published: October 2019

Abstract
The risk for donors of allogeneic hematopoietic stem cells (HSC) by bone marrow (BM) or by peripheral blood (PB) harvest is generally considered negligible. Scattered reports of severe to life-threatening complications and a recent controversy on hematopoietic malignancies after GCSF administration for peripheral stem cell donation have challenged this opinion. Previous studies were limited by small numbers. In two consecutive retrospective surveys conducted in 2003 and 2006 amongst 338 allogeneic transplant centres from 38 European countries participating in the annual EBMT activity surveys, centres were asked to report all donor deaths, all severe adverse events (SAE’s), defined as occurring within 30 days and any hematological malignancy in a donor occurring after HSC donation. 262/338 teams (77.5%) responded to the first survey (1993–2002) and 169/262 (65%) centres replied to the second survey (2003–2005). The responding teams performed a total of 51’024 first allogeneic HSCT, 27’770 BM and 23’254 PB HSCT, which corresponds to 69% of all 73’947 first allogeneic HSCT reported during this time to EBMT. There were 5 donor deaths, 1 after BM and 4 after PB donation, an incidence of 0.98 per 10’000 donations (95% CI 0.32–2.29), 37 SAE’s (incidence 7.25/10’000 donations; 95% CI 5.11–9.99), 12 in BM (incidence 4.32/10000 donations; 95% CI 2.24–7.75) and 25 in PB donors (incidence 10.76/10’000 donations; 95% CI 6.97–15.85; p<0.02). In absolute numbers, there were 20 hematological malignancies occurring in donors (3.92/10’000 donations; 95% CI 2.39–6.05), 8 after BM (2.88/10’000 donations; 95% CI 1.24–5.68) and 12 after PB donation (5.16/10’000 donations; 95% CI 2.67–9.02; p = 0.3). Based on the different observation times, the incidence rates for developing hematological malignancies are 0.398 per 10’000 person-years for BM and 1.20 per 10’000 person-years for PB donation, resulting in a relative risk of 3.02 (95% CI 1.11–6.87, p=0.027). These data document a definitive risk for death, SAE’s and hematological malignancies with HSC donation. Deaths occur with similar frequency in both groups. SAE’s were more frequently reported after PB donation. The incidence rate for developing hematopoietic malignancies is higher after PB donation. These data clarify the recent controversy on HSC donation. They form a basis for donor counselling and underline the need for standardised donor follow up and international cooperation in order to define risk factors and to build up preventive measures.
Pegfilgrastim in Congenital Neutropenia: An Observational Survey from the French SCN Registry.
Authors: Beaupain B. , Leblanc T. , Reman O. , Vannier J.P. , Hermine O. , Suarez F. , Bertrand Y. , Bellannre-Chantelot C. , Donadieu Jean 
Published: October 2019

Abstract
PegFilgrastim is registered in France since 2002. Its pharmacokinetic characteristic allows the possibility to limit the number of injections in patients with congenital neutropenia receiving long term G-CSF therapy. To date, no report has been issued about the outcome of patients with congenital neutropenia receiving this drug. At the cut-off date of July 31th, 2007, 339 patients with congenital neutropenia had been included in the French Severe chronic neutropenia (Severe congenital neutropenia n=142, WHIM n=16, Cyclic n=75; GSDIb n=16 SDS n=90). Eight patients (SCN n=4, Cyclic n=3 and GSDIb n=1) had been identified to receive PegFilgrastim. Median age at PegFilgrastim onset was 18.2 years (min 6.1-max 37.6). In 7 cases, PegFilgrastim was given in the continuation of Filgrastim (n=3) or Lenograstim(4), with a median delay of 13 years after G-CSF onset and in one case, PegFilgrastim was the initial prescription of cytokine. Dose of PegFilgrastim was usually a vial per injection, except in one child (1/2vials) resulting in a dose per injection between 90 and 286 μg/kg per injection. The rhythm of PegFilgrastim injection varied from 1 injection every 7 days until 1 injection every 14 days, with discontinuation period. The median duration (between onset of PegFilgrastim and the last dose) was 0.7 year (min 0.3–2.7). The efficiency measured by ANC criteria appears to better than G-CSF (fig 1: baseline Median ANC 156/mm3, Under G-CSF 501/mm3, Under PegFilgrastim 1901/mm3/ Kruskall Wallis test p=0.02). Severe infection was never reported under PegFilgrastim, but neither in the previous G-CSF therapeutic periods for the 7 patients. More frequent ENT infection was observed in one patient who consider the PegFilgrastim as less efficient than G-CSF. The side effects appeared to be more frequent compare to the G-CSF therapy. Bone pains were reported in 7 patients, anemia and thrombocytopenia (WHO gr II) in one, urticarian chronic skin rash in 3. Finally, at the last update, 7 patients had withdrawn PegFilgrastim, and had resumed G-CSF therapy. Only one patient is still receiving PegFilgrastim. In conclusion, PegFilgrastim appeared efficient in Congenital neutropenia, but more frequent short term side effects were observed compare to standard G-CSF therapy, resulting in a limited quality of life improvement, leading to drug withdrawn in most of the patients.
Figure 1: Median ANC by diagnosis Figure 1:. Median ANC by diagnosis
Reducing MCM Loading Causes Chromosomal Aneuploidy.
Authors: Orr Stephen J. , Gaymes Terry , Wang Rong , Czepulkowski Barbara , Ladon Darius , Chronis Constantinos , Mufti Ghulam J. , Marcotte Edward M. , Thomas N. Shaun B. 
Published: October 2019

Abstract
Normal DNA replication must be accurate and occur only once per cell cycle. Sites of DNA replication are specified by binding the origin recognition complex, that includes minichromosome maintenance (MCM) proteins. Paradoxically, in higher eukaryotes MCM proteins are present in >20 fold excess of that required for DNA replication. They are also downregulated by elevated expression of proteins such as cyclin E that occurs in cancers, including AML and breast cancer. We investigated why human cells need “excess” MCM proteins and whether the reduction of MCM protein levels might contribute to a malignant phenotype. We determined the consequences of reducing the levels of MCM proteins in primary human T cells in which cell cycle controls and DNA damage responses are normal. Mass spectrometry sequencing of chromatin/nuclear matrix-bound proteins and western blotting identified that Mcm7 is not present in quiescent, normal primary human T cells. Mcm7 is induced in mid G1after the G0→G1 commitment point, the point beyond which T cells are committed to entering the cell cycle. Reduction of Mcm7 with siRNA to <5% of normal during G0→G1→S-phase reduces chromatin-binding of each of the MCM proteins that form the DNA helicase. However, these cells still enter S-phase and replicate DNA. Reducing MCM levels by titrating siRNA causes dose-dependent DNA-damage responses involving activation of ATR & ATM and Chk1 & Chk2. However, cells depleted of Mcm7 do not undergo apoptosis, rather reducing MCM levels even by 50% causes gross non-clonal chromosomal abnormalities normally found in genomic instability syndromes. M-FISH identified chromosome translocations, as well as loss and gain of individual chromosomes, which can occur individually or together in the same cell. Reducing MCM levels also causes misrepair by non-homologous end joining (NHEJ), and both NHEJ and homologous recombination (HR) are necessary for chromosomal abnormalities to occur. Therefore, “excess” MCM proteins that are present in a normal, proliferating cell are necessary for maintaining genome stability and reduction of MCM loading onto DNA that occurs in cancers is sufficient to cause genomic instability.
Leukemic Promyelocytes Possess Self-Renewal Capacity and Leukemia Stem Cell Properties in a Mouse Model of Acute Promyelocytic Leukemia.
Authors: Wojiski Sarah A. , Lee Benjamin , Kindler Thomas , Gozo Maricel , Leeman Dena , McDowell Elizabeth , Fabian Attila , Guibal Florence , Tenen Daniel , Gilliland D. Gary 
Published: October 2019

Abstract
It has been determined that within the heterogeneous leukemic population, there is a rare subset of leukemia stem cells that are responsible for continued maintenance and propagation of the tumor. Recent evidence suggests that during disease pathogenesis, more mature progenitor cells are capable of acquiring stem cell-like characteristics including properties of self-renewal and the capacity to differentiate into progeny that lack self-renewing potential. Acute promyelocytic leukemia (APL) is associated with the t(15;17)(q22;q12) chromosomal translocation that produces the PML-RARa fusion protein, and is unusual among the various subtypes of AML in that it is characterized by hyperproliferation of differentiated promyelocyte progenitors. In a mouse model in which PML-RARa is expressed under the control of the endogenous murine cathepsin G promoter, we observed that whole bone marrow expressing PML-RARa derived from pre-leukemic mice has certain properties of self-renewal, such as serial replating potential in methylcellulose cultures, and serial transplantation of disease into recipient animals. Using multiparameter flow cytometry, we demonstrated that preleukemic PML-RARa animals have an expansion of mature Gr-1+c-kit-CD34− granulocytes that are not capable of self-renewal. However, in the leukemic state, the expanded population shifted to a more immature population with an immunophenotype of Gr-1+c-kit+CD34+. This population was capable of transplanting disease to secondary recipients that recapitulated the disease of the primary donor, and thus contained leukemia-initiating activity. Morphologically, these cells resembled promyelocytes, and their promyelocytic identity was confirmed by quantitative PCR detection of the expression of azurophilic primary granule components associated with the promyelocyte stage of development, including myeloperoxidase, neutrophil elastase, and cathepsin G. However, these cells lacked expression of the secondary granule protein gelatinase B associated with the more mature myelocyte stage of granulocytic differentiation. Additionally, these leukemia-initiating promyelocytes responded to differentiation and cell death signals mediated by administration of all-trans retinoic acid (ATRA). These data indicate that in APL, a highly differentiated promyelocyte compartment does in fact possess properties of leukemia stem cells. This suggests that an important step in disease pathogenesis mediated by PML-RARa is the acquisition of properties of self-renewal in a terminally differentiating progenitor that normally lacks this potential.