bluuebunny/crossref_metadata_2025_split huggingface dataset:
Who Gets Inhibitors? Predicting Inhibitory Antibodies in Children with Severe Hemophilia A.
Authors: ter Avest Petra C. , Fischer Kathelijn , Santagostino Elena , van den Berg Marijke H. , van der Bom Johanna G. 
Published: October 2019

Abstract
Background. Replacement therapy in severe hemophilia A patients is complicated by formation of inhibitors in around 25% of children. Identification of patients at the highest risk may help to tailor personalized treatment strategies.
Objectives. To develop a scoring system that may be used to identify patients at the highest risk of inhibitor development at first treatment.
Methods. We used the data from a retrospective multicentre cohort study (the Canal cohort) of patients with severe hemophilia A (factor VIII (FVIII)less than 0.01 IU/ml), born 1990–2000, followed at least until their 50th exposure day. Presence of inhibitor was defined as twice a positive inhibitor titer and a decreased FVIII recovery. Based on the coefficients of a logistic regression model, a weighted risk-score was developed. Shrinkage of regression coefficients was used to control for overfitting. The discriminative ability of the risk-score was expressed as the area under the curve (AUC) of a receiver operating characteristic curve.
Results. Out of the 366 patients from the Canal cohort, 284 children were selected of whom 78 developed an inhibitor (27%). Logistic regression analysis revealed 3 independent risk factors for inhibitor development: positive family history, intensive treatment (at least 5 consecutive days) at the first FVIII exposure, and high risk FVIII gene mutations. Table 1 presents odds ratios of the uni- and multivariate analyses, and the risk score. The AUC for the risk-score was 0,724. Table 2 shows that the model is able to separate high and low risk patients from patients with an intermediate risk of 25%.
Conclusions. The risk score includes positive family history of inhibitors, high risk factor VIII gene mutation and intensive treatment at first FVIII exposure. This risk score can recognize patients with a doubled risk for inhibitor development and may be used to guide inhibitor preventive treatment strategies.
Table 1. Uni- and Multivariate analysis and risk-score. OR (CI), Univariate OR (CI), Multivariate p-value Risk-Score CI = confidence interval 95% Positive Family History of Inhibitors 4.0 (1.7–9.4) 3.0 (1.2–7.7) ,022 3 High risk FVIII Gene Mutation 3.6 (1.8–7.2) 3.7 (1.8–7.9) ,001 4 Intensive Treatment at 1st FVIII exposure 6.8 (3.6–12.9) 7.2 (3.4–15.1) ,000 6 Table 2. Calibration of the risk-score. Model Total n° patients Predicted n° Inhibitors Observed n° Inhibitors Positive Predictive Value Negative Predictive Value LR= Low Risk, MR= Medium Risk, HR= High risk LR:0 72 7 6 0,34 0,92 MR:3–4 153 39 38 0,25 0,69 HR: >4 59 32 34 0,58 0,80
Effects of Recombinant Factor VIIa (NovoSeven®) on Restoring Thrombin Generation in Patients with Hemophilia A and Antibodies to Factor VIII.
Authors: Eichinger Sabine , Lubszcyk Barbara , Zwiauer Karl , Gleiss Andreas , Quehenberger Peter , Kyrle Paul Alexander 
Published: October 2019

Abstract
Introduction: Development of antibodies (ab) against factor VIII (FVIII) is a serious complication of replacement therapy in patients with hemophilia A. In case of bleeding patients with FVIII ab are treated with agents that induce hemostasis independently of FVIII. Recombinant activated factor VIIa (rVIIa) shows clinical efficacy, but its effects on hemostatic system need still to be fully elucidated.
Methods: In an open controlled study, we investigated thrombin generation (peak thrombin) and parameters of coagulation activation [D-Dimer, prothrombin fragment F1+2 (F1+2)] in 5 patients with hemophilia A and FVIII ab, and in 5 healthy age-matched controls before and after intravenous bolus infusion of rVIIa (90 μg/kg bodyweight NovoSeven®, NovoNordisk, Denmark) (in hemophiliacs only). All parameters were measured in plasma before and 0.5, 1, 2, 3, and 4 hours after rVIIa infusion by use of commercially available assays (Technothrombin®TGA, Technoclone, Austria; Asserachrom®D-Di, Diagnostica Stago, France; Enzygnost F1+2, Dade Behring, Germany).
Results: At baseline, hemophilia A patients had markedly lower mean (min-max) peak thrombin levels than controls [0.12 (0.0–0.6) nM vs. 186.9 (116.0–254.4) nM]. Mean (min-max) F1+2 levels did not significantly differ between patients and controls [160.7 (89.8–331.3) pmol/l vs. 160.8 (104.4–242.3) pmol/l]. Notably, D-Dimer levels were significantly higher in hemophiliacs than controls [1087.5 (174.8–3882.4) ng/ml vs. 146.3 (87.2–289.8) ng/mL]. FVIIa levels reached a mean (min-max) maximum of 28 (24–32) U/ml after 0.5 hours in all patients. After infusion, a considerable increase in mean (min-max) peak thrombin levels to 40.7 (28.3–51.6) nM was seen. Time to maximum levels was 30 minutes in three patients and 60 minutes in two. For each of the five patients the peak thrombin level was substracted from the level of its matched control at the same time point. The mean of these differences was 168.7 nM (95% CI 82.6–254.8), which translates into 80.2% (95% CI 65.4% – 88.6%) lower peak thrombin levels in haemophiliacs with FVIII ab. F1+2 significantly increased in all patients [mean (min-max) maximum levels 292.5 (175.1–464.3) pmol/l]; time to maximum levels varied from 2 to 4 hours. D-Dimer levels remained almost unchanged in all patients.
Conclusion: Patients with hemophilia A and FVIII ab have low in vitro thrombin generation and F1+2 levels. After rVIIa infusion, coagulation activation as measured by F1+2 levels is slightly increased, and thrombin generation capacity is restored by 20% compared to healthy controls. Measurement of peak thrombin could be useful to monitor procoagulant treatment of patients with hemophilia A and FVIII ab.
The Activity and Toxicity of Dasatinib in Relapsed or Plateau-Phase Multiple Myeloma.
Authors: Wildes Tanya M. , Ruddell Alisa , Gao Feng , Uy Geoffrey , Cashen Amanda , Abboud Camille , Westervelt Peter , Dipersio John F. , Vij Ravi 
Published: October 2019

Abstract
Dasatinib (Sprycel®) is an orally administered inhibitor of multiple tyrosine kinases, which has been approved for the treatment of CML in chronic phase, accelerated phase or blast phase with resistance or intolerance to prior therapy including imatinib. In vitro studies have demonstrated that dasatinib inhibits the viability of multiple myeloma cell lines. Based on the in vitro activity of dasatinib against multiple myeloma, we initiated an open label, single-arm Phase II trial of dasatinib in relapsed, refractory or plateau-phase multiple myeloma; planned enrollment is 21 patients (pts). Treatment consists of dasatinib 70 mg p.o. twice daily continuously. The primary endpoint of the study is the overall response rate (CR+PR). Secondary endpoints include the time to response, duration of response, time to progression as well as tolerability and safety of dasatinib in this population. To date, 13 pts have been enrolled. The median age is 58 (range 38–76). Stage at diagnosis was: IIA (1 pt), IIIA (11), IIIB (1); 9 pts had IgG paraprotein and 4 had IgA. Median beta2-microglobulin at study entry was 3.2 (range 1.2–13). The median number of prior therapies was 2 (range 1–14); 12 pts had undergone prior autologous stem cell transplant. At enrollment, 7 pts had progressive disease and 6 pts had plateau phase disease, defined as at least 3 months with a stable paraprotein level (±25%) following the most recent therapy. At a median follow-up of 39 days (range 1–134 days), results are as follows: 4 patients have stable disease (SD); 7 patients had progressive disease (PD); 1 pt discontinued therapy due to toxicity (Grade II diarrhea, flushing and fatigue); 1 pt is not yet evaluable for response. Of those who remain on therapy, the median time on therapy is 39 days (range 1–134); 4 of these 5 patients had plateau phase disease at enrollment. Of those who discontinued therapy for PD, median time on therapy was 46 days (range 15–91); 6 of these 8 patients had PD at the time of enrollment. Toxicities have been manageable. Hematologic toxicities were more pronounced in the patients who had cytopenias at baseline. There was 1 episode of Grade III neutropenia, 4 episodes of Grade III anemia, 2 episodes of Grade III thrombocytopenia, and 3 episodes of Grade IV thrombocytopenia. Grade III non-hematologic toxicities have included: pneumonia (3 pts), acute renal failure (2), headache (1), epistaxis (2), febrile neutropenia (1), pulmonary edema (1), GI bleed (1), fatigue (2), hypertension (1), and hip pain (1). There was 1 episode of grade IV hypoglycemia. Enrollment and evaluation of responses/toxicities are ongoing. The protocol is being amended to allow for dose escalation to dasatinib 100 mg b.i.d. for stable disease at 8 weeks on therapy.
In Vivo Expansion of Megakaryocyte Progenitors in CD26−/ − Mice: Implications for Novel Post-Transplant Therapeutics.
Authors: Kidd Shannon , Bueso-Ramos Carlos E. , Paganessi Laura A. , Fung Henry , Gregory Stephanie , Christopherson Kent W. 
Published: October 2019

Abstract
Hematopoietic stem cell transplantation (HSCT) is a successful treatment option for patients with malignant or non-malignant severe hematologic diseases. Readily available umbilical cord blood (CB) has emerged as an important donor source that has a lower histocompatability requirement and carries a reduced risk of graft vs. host disease. However, a frequent complication of CB use is delayed hematopoietic recovery, in particular long lasting severe thrombocytopenia (median of 117 days to platelet recovery, COBLT 2005). To overcome this clinically, effective strategies for enhancing megakaryopoiesis, thrombopoiesis, or both are needed. We have previously described the importance of CD26 (dipeptidylpeptidase IV) in engraftment (Christopherson, KW 2nd, et al, Science2004. 305:1000–3). However, the involvement of CD26 in megakaryopoiesis has not been investigated. We hypothesized that CD26 acts to suppress megakaryopoiesis and that removal of CD26 activity would result in expansion of the megakaryocyte progenitor population in vivo. To test this hypothesis, we evaluated megakaryocyte (MK) development in the context of development of other mature blood cells in CD26 deficient (CD26−/ −) mice as compared to control C57BL/6 mice. Histological analysis of formalin fixed paraffin embedded tissue sections and peripheral blood cytopsins revealed an increased presence of MK in the bone marrow, spleen, thymus and peripheral blood. However, complete blood counts (CBC) suggest no difference in white blood cell, neutrophil, lymphocyte, monocyte, eosinophil, basophil, and platelet counts. Flow cytometric analysis also revealed no significant changes in CD3, CD4, or CD8 T-cells; B220 B-cells; and Gr-1/Mac-1 granulocytes/neutrophils in the bone marrow, spleen, thymus, or peripheral blood as appropriate. There was also no change in the percentage of peripheral blood CD41 MK but there was a increase from 0.09% in C57BL/6 BM to 0.26% CD41+Sca-1+c-kit− MK progenitors in the CD26−/ − BM (P≤0.05). Methylcellulose based myeloid progenitor assays did also reveal respective CFU-GM, BFU-E, and CFU-GEMM per femur values (mean±SEM) of 24480±1426, 1448±154.86, and 852±41 for C57BL/6 BM cells; 24451±1342, 974.29±81, and 1634±177 for CD26−/ − BM cells. This represents a 92% increase in CFU-GEMM and corresponding 30% decrease in BFU-E in the CD26−/ − mouse BM (P≤0.01, n=7 mice/group). Collagen based megakaryocyte progenitor assays (CFU-MK) revealed a 25% increase from 4455.00±207.62 colonies/femur in C57BL/6 BM to 5555.00±608.12 in CD26−/ − BM (P≤0.05, n=3 mice/group). These results establish a basis on which to propose that CD26 may act to regulate early events in megakaryocyte progenitor formation and function. They also suggest that the use of CD26 inhibitors may have a beneficial effect on improved megakaryocyte progenitor function and/or reconstitution post-transplant.
Ex Vivo Treatment of Human Cord Blood with dmPGE2 Can Safely Increase the Potential for Hematopoietic Engraftment.
Authors: North Trista E. , Goessling Wolfram , Armant Myriam , Kao Grace S. , Silberstein Leslie E. , Ritz Jerome , Zon Leonard I. 
Published: October 2019

Abstract
Hematopoietic stem cells (HSCs) are commonly used in transplantation therapy to rescue the hematopoietic and immune systems following systemic chemotherapy or irradiation. However, some patients receive inadequate numbers of HSCs and this often results in delayed reconstitution of hematopoiesis and immune function and associated toxicities. We previously demonstrated that a stabilized derivative of prostaglandin (PG) E2 increases vertebrate HSCs both in vivo and in vitro. 16,16-dimethyl PGE2 (dmPGE2) significantly increased HSCs during zebrafish embryogenesis and in the adult marrow following injury. Incubation of murine embryonic stem cells with dmPGE2 during embryoid body differentiation resulted in a dose-dependent increase in hematopoietic colonies, demonstrating that the function of PGE2 in HSC regulation is conserved in mammals. Finally, ex vivo treatment of murine bone marrow with dmPGE2 resulted in a 2-fold increase in engrafting cells in a limiting dilution competitive repopulation assay. No negative effects on serial transplantability of HSCs were observed in these animal models. To investigate the therapeutic potential of PGE2 for the amplification of blood stem cells, we exposed human cord blood (hCB) cells to dmPGE2 in vitro and measured the effects on stem and progenitor populations both in vitro and in vivo. Red cell depleted umbilical cord blood specimens, cryopreserved for clinical use, were thawed and divided for parallel processing. Ex vivo treatment of hCB cells for 1 hour with dmPGE2 in dextran/albumin had no negative impact on absolute cell count or the viability and relative distribution of both CD45 and CD34 positive cells compared to vehicle treated control hCB cells. Significantly, hCB treated with dmPGE2 produced enhanced numbers of GM and GEMM colonies in methylcellose CFU-C assays compared to controls. Human CB cells treated ex vivo with dmPGE2 for 1 hour and transplanted at a dose of 20 million live CD45+ cells per recipient were capable of repopulating NOD/SCID mice after sublethal irradiation. In comparative studies at 6 weeks post transplantation, human CD34+ and CD45+ cells could be detected in the marrow (>2%) of dmPGE2 treated (4/8) and control treated (1/6) recipients. Long-term and competitive transplantation experiments to assess the effect of dmPGE2 treatment on functional HSCs are currently in progress. Our data suggests that treatment of human cord blood products with dmPGE2 will be both safe and effective in achieving expansion of hematopoietic stem cells for transplantation in the clinical setting. TE North and W Goessling contributed equally to this work.
Exchange of microRNAs between Breast Cancer Cells and Bone Marrow Stroma: Potential for Avenues To Revisit Hematopoietic Stem Cell Transplantation for Breast and Other Cancers That Show Preference for Bone Marrow.
Authors: Taborga Marcelo , Rameshwar Pranela 
Published: October 2019

Abstract
Previous failure of autologous bone marrow transplantation as a viable treatment of breast cancer opens the field for investigational studies since the bone marrow, as a source of cancer metastasis, remains a clinical dilemma. This study hypothesizes that a subset of breast cancer cells enter the bone marrow at an early period of disease development, perhaps prior to clinical detection, where they evade chemotherapeutic damage even at high doses. An understanding of this cell population might be crucial for the future success of bone marrow transplants in patients suffering from breast cancer metastasis to the bone marrow. In vivo and in vitro studies have demonstrated gap junctions between bone marrow stroma and breast cancer cells. The gap junctions can passage CFDA cell tracer between stroma and cancer cells. This exchange can be blocked by the gap junctional intercellular communication inhibitor, 1-Octanol, suggesting functional gap junctions. We have now implemented quantitative studies to determine the role of specific microRNAs (miRNAs) in blunting of mRNA translation of the Tac1 gene in cancer cells. The first set of studies optimized the effects of miRNAs, 130a and 206 in the suppression of Tac1 mRNA in the low invasive T47D and the aggressive MDA-MB-231 breast cancer cell lines. The miRNAs were transfected into cells containing the pMIR-R/Tac1/SG vector, which includes the 3′ UTR of Tac1 mRNA. Luciferase activity in cell lysates showed synergism between the two miRNAs, indicating that the predicted miRNAs can inhibit Tac1 translation. The next set of studies determined whether miRNAs 130a and 206 can cross gap junctions between stroma and cancer cells to suppress Tac1 mRNA. The miRNAs were transfected in one cell type and pMIR-R/Tac1/SG in the other cell type. Luciferase activity and parallel studies with 1-Octanol verified movement of miRNAs via the gap junction. These studies recapitulate the downregulation of endogenous Tac1 and other cytokines in the cancer cells. In summary, this study shows a role for miRNA exchange, between bone marrow stroma and breast cancer cells, as a mechanism by which the cancer cells might evade detection and current methods of treatment. An understanding of these mechanisms might lead to new methods to remove cancer from bone marrow in future therapies.
Large Regions of Uniparental Disomy (UPD) Establish Clonal Hematopoietic Stem Cell Selection in a Subset of Myelodysplastic Syndrome (MDS) Patients with Normal Bone Marrow Cell Karyotypes.
Authors: Heinrichs Stefan , Kulkarni Rima V. , Bueso-Ramos Carlos E. , Loh Mignon , Levine Ross , Li Cheng , Neuberg Donna S. , Kornblau Steve M. , Issa Jean-Pierre , Gilliland D. Gary , Estey Elihu H. , Look A. Thomas 
Published: October 2019

Abstract
Myelodysplastic syndromes (MDS) comprise a large group of hematological diseases sharing clinical features, but the diverse mechanisms underlying their molecular pathogenesis remain largely undefined. In this study, we used high-density 250K SNP arrays to analyze matched constitutional (buccal) and whole bone marrow DNAs from 36 patients with MDS. In 14 cases, we also selected CD34+ stem and progenitor cells for comparison. 23 of the cases were cytogenetically normal. Whereas regions of genomic amplification were rare, we found distinct regions of heterozygous deletion in 9 patients, most frequently targeting chromosomes 5q, 7q, 17p and 20q. In one patient with a large heterozygous 5q deletion, a smaller region in 5q34 was homozygously lost. Based on genotyping we could identify regions of uniparental disomy (UPD) in 4 patients, three of whom had normal bone marrow karyotypes. Two of the regions of UPD targeted chromosome 7q, suggesting that the frequency of 7q deletion is underestimated by conventional cytogenetics, and that a mutation has occurred in these cases within the reduplicated region involved in UPD. Comparison with constitutional DNA was critical for accurate SNP analysis in MDS, allowing us to focus on acquired clonal abnormalities and control for changes due to inherited copy number variation or unusual regions of germline homozygosity. For example, one case had several large blocks of germline homozygosity distributed over various chromosomes, and by comparison with the buccal DNA we were able to discern that these regions did not result from clonal somatic abnormalities associated with MDS. The analysis of CD34+ cellular DNA in 14 cases revealed that all aberrations identified in the stem and progenitor fraction were also identified in whole bone marrow DNA. This finding emphasizes the dominance and stem cell involvement of the abnormal clone in MDS, which makes purification of CD34 cells unnecessary for SNP array analysis in this disease. Most, but not all, clonal deletions evident by cytogenetic analysis were detected as copy number alterations by SNP array, possibly indicating that such abnormalities occurred in subpopulations of the cytogenetically analyzed MDS cells. Thus, SNP array and cytogenetic analysis are complementary methods for the identification of abnormal clones in MDS patients, each detecting overlapping but distinct subsets of patients. Our data indicate that SNP array analysis should be performed in all patients with MDS and normal bone marrow cytogenetics, in order to help discriminate patients with UPD indicating cell autonomous clonal stem cell malignancy from those who might have extrinsically mediated bone marrow failure syndromes.
Evaluation of an Automated Ficoll Procedure To Enrich for Mononuclear Cells for Use in Clinical Trials.
Authors: Sepulveda Carmen A. , Richman Sara J. , Kaur Indreshpal , Shen Anna , Khorana Sangeeta , O’Connor Sean L. , Champlin Richard E. , Shpall Elizabeth J. , McMannis John D. 
Published: October 2019

Abstract
Mononuclear cell fraction (MNC) enrichment from hematopoietic progenitor cells (HPC) cells is required for a number of clinical applications. Previously, we have used a manual method for enrichment of these cells. We evaluated an automated procedure for better consistency and patient safety (reduce risk of contamination). We have been using the Sepax (BioSafe) for the past 18 months to process Cord Blood (RBC depletion) before cryopreservation. Since the instrument can also do density gradient enrichment of mononuclear cells, we evaluated the ability to process different sources of hematopoietic progenitor cells (HPC) on the Sepax.
Methods: Bone marrow, non-mobilized apheresis and cord blood were evaluated as a source of HPC. Briefly, 100ml of Ficoll solution were added to the chamber of the Sepax disposable. After initiating centrifugation, 50–110 ml of HPC were then overlayed onto the density gradient. The cells were fractionated, mononuclear cell fraction isolated, and then washed three times before final resuspension in 50ml of Saline/5% albumin. In some experiments the HPC were divided into two fractions so that a direct comparison of the Manual vs Automated enrichment procedure could be compared. Finally, a recent publication from Seeger et al (European Heart Journal28:766, 2007) suggested different density gradient solutions have an effect on MNC enrichment. Therefore, four different brands of density gradient solution (GE Ficoll-Paque Premium, Sigma Histopaque 1077, Accurate Lymphoprep and Lonza Lymphocyte Separation media) were evaluated to determine if there was a differential separation of subsets of cells.
Results: The total nucleated cell (TNC), MNC and CD34 recoveries after MNC enrichment were similar for either bone marrow or cord blood HPC comparing the manual procedure and the automated procedure. The TNC and MNC recovery was higher for apheresis products (66% and 69% respectively) than the other two sources of HPC. This most likely reflects the enriched MNC starting population from an apheresis device. We evaluated the TNC and MNC recoveries over a range of initial TNC (7.5–20 × 108 cells) but could not see an effect of either cell number or cell concentration on recoveries. Nor could we see an effect of initial cell volume (50–100 mls) on recovery of cells post processing. Finally, we could not see a difference in either TNC or MNC recovery when the four different sources of density gradient solution were compared.
Conclusion: We have shown that the Sepax instrument can be used for enrichment of mononuclear cells from HPC, Marrow, HPC, Apheresis and HPC, Cord. The benefits of an automated system are
consistency, functionally closed system which reduces possibility of contamination documentation of run parameters and shorter processing time.