bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genetic Modeling of Chemical Antagonists: Genome Scale Validation of Hematopoietic Disease Drug Targets by In Vivo Functional Analysis.
Authors: Oravecz Tamas , Turner Alex , Brown Philip , Zambrowicz Brian , Blobel Gerd 
Published: October 2019

Abstract
Clinical management remains elusive for a large number of patients with hematological and other disorders necessitating the development of new, more effective pharmaceutical agents. While the sequencing of the human genome has provided a list of all drug targets that will ever exist, in order to improve the speed and cost of drug development there is a critical need to identify the small subset of these targets whose modulation will lead to therapeutic outcomes. As a genetic model of a perfectly selective and potent chemical antagonist, the analysis of the phenotypes of knockout mice allows one to predict the mechanism based efficacy and side effect profile of modulators of a particular target in a physiologic mammalian setting without having to first develop a candidate drug with which to validate the target. The power of this approach has been demonstrated by studies examining the targets of the best-selling drugs and current pharmaceutical pipelines. These analyses have confirmed a strong (~85%) correlation between the target’s knockout phenotype and the efficacy and/or side effects of drugs that modulate them. We have implemented this target validation approach by developing systems and infrastructure to generate and comprehensively phenotype mutant mice at a rate of more than fifteen novel lines per week. The battery of tests encompassed by our analysis have been specifically selected to reveal those genes that encode key control points in mammalian physiology whose modulation may be used to treat major disease processes including hematologic disorders. Tests to detect hematologic phenotypes include complete blood count; immunophenotyping; blood chemistry; analysis of soluble factors (immunoglobulins, cytokines and inflammatory mediators); full histopathologic examination, and various in vivo and in vitro immune challenge and cell activation assays. In addition, selected lines are subjected to more extensive analyses including additional challenge assays, gene expression profiling, and bone marrow transplantation. Data will be presented describing one of the lines analyzed in this screen, the Abl2 knockout, which has an increased mean total white blood count relative to wild-type littermates, as well as smaller body size (decreased length and weight). In parallel with the identification of therapeutically relevant proteins, we are actively exploring pharmacologic methods, both with small molecules and therapeutic antibodies, to modify the function of these novel gene targets, and ultimately to treat human disease.
Association between Statin and Fibrate Use and Venous Thromboembolism: A Case-Control Study.
Authors: Lacut Karine , Le Gal Gregoire , Oger Emmanuel , Mottier Dominique 
Published: October 2019

Abstract
Background:
Previous studies of selected patients have suggested a reduction in the risk of venous thromboembolism (VTE) with the use of statins, and no effect of fibrates.
Objective:
To evaluate the influence of statin and fibrate use on the risk of venous thromboembolic events.
Design:
Case-control study (EDITH) designed to investigate genetic and environmental risk factors of VTE.
Setting:
Brest University Hospital.
Participants:
857 patients consecutively hospitalized for a documented venous thromboembolic event were included between May 2000 and May 2004. Controls were matched on age, sex and the main risk factors of VTE (cancer, surgery, pregnancy…).
Results:
The mean age of patients was 67.7 year. No significant difference was found between cases and controls concerning the main characteristics, except for smocking and body mass index. Controls had more often previous vascular events (coronary heart disease, stroke or arteriopathy of the lower limbs) than cases but the difference was no significant. Statin use was associated with a significant decreased risk of VTE (odds ratio (OR) = 0.58; 95% confidence interval (CI), 0.41–0.82), whereas fibrate use was associated with a significant increased risk of VTE (OR = 1.60; 95% CI, 1.09–2.34). After adjustment on the main confounding factors including aspirin use and cardiovascular disease, these associations remained significant. Among pleiotropic effects of statins, some antithrombotic mechanisms could be proposed to explain their possible protective effect. Concerning the possible negative effect of fibrates, some authors found that the most prescribed fibrates, but not statins, caused hyperhomocysteinemia. In our study, analyses of homocysteinemia are ongoing.
Conclusion:
In this case-control study of hospitalized patients, statin use was associated with a significant decreased risk of VTE, whereas fibrate use was associated with a significant increased risk. Homocysteinemia may be involved in the difference between the effects of these two categories of lipid-lowering drugs on VTE. Because our study was observational, the protective effect of statins as regards the risk of VTE remains questionable and further prospective studies are needed.
The Risk of Venous Thromboembolism (VTE) among Cancer Patients by Tumor Site: A Population-Based Study.
Authors: Heit John A. , Petterson Tanya M. , Marks Randolph S. , Bailey Kent R. , Melton L. Joseph 
Published: October 2019

Abstract
Active cancer is an independent VTE risk factor (overall 6- to 9-fold increased risk) and accounts for almost 20% of all VTE in the community, but which cancer patients are at risk for VTE is largely unknown. Reportedly, VTE risk varies by tumor site, and cancer of the ovary, pancreas, colon, stomach, lung, prostate, and kidney convey particularly high VTE risk.
Objective: To estimate VTE risk by tumor site.
Methods: We enumerated observed cancers by tumor site for Olmsted County, MN active cancer patients with incident VTE over the seven-year period, 1991–1997 (n=152). We used 1991–1997 State Surveillance, Epidemiology, and End Results (SEER) data for Iowa to estimate the expected age-specific prevalence of cancer by tumor site in Olmsted County. VTE risk ratios (RR) for each tumor site were estimated by dividing the observed number of cancers by the expected number (calculated as the product of the SEER prevalence and the number of incident VTE cases in the age stratum).
Results: For our population of 1991–1997 VTE cases, all tumor sites had RR > 5.0 (range 5.2 to 37.3, all p-values<0.05). Compared to published overall VTE odds ratios of 6–9 for active cancer compared to no cancer, the RR for some tumor sites were particularly increased. A Chi-squared test of heterogeneity of the RR across sites was highly significant (p-value<0.001). Three rare cancer sites - pancreatic cancer, lymphoma, and brain cancer - had unusually high RR (all RR>25). The high number of VTE cases with lymphoma was not due to catheter-related arm vein thrombosis. Liver, leukemia, other gastrointestinal (esophagus, small intestine, gallbladder, other biliary) and other gynecologic (primarily cervical) cancers had over twice the baseline risk (i.e., RR>17.0). On the other hand, the RR for many common cancers (breast, colorectal, ovary, lung, prostate) were essentially the same as the overall baseline risk (all had 9.5<RR<12.0).
Conclusions: In contrast to previous reports, pancreas, lymphoma, brain, liver, leukemia, other gastrointestinal, and other gynecologic cancers have the highest VTE risk. Prior estimates of VTE risk by tumor site may have been biased by studies of prevalent cancers among patients hospitalized in tertiary care centers.
Thrombohemorrhagic Defects and Recurrent Miscarriage Syndrome.
Authors: Bick Rodger L. , Hoppensteadt Deborah. 
Published: October 2019

Abstract
We have assessed 351 women referred for thrombosis & hemostasis evaluation after suffering recurrent miscariages. All patients were referred by a high-risk obstetrician or reproductive medicine specialist after anatomic, hormonal or chromosomal defects had been ruled out. These patients were assessed over a three year period. The mean patient age at referral was 34 yrs. and the mean number of miscarriages was 3.4 (2–9). All patients underwent a thorough evaluation for thrombophilia and, when indicated, a hemorrhagic disorder. Of the 351 patients, 29 (8%) had no defect. Of the remaining 322 patients, 7 (2%) had a bleeding disorder: 3 with platelet dysfunction, 1 with Factor XIII deficiency, 3 with von Willebrands and 3 with Osler-Weber-Rendu. The remainder of the patients had a thrombophilia as follows: 195 (60%) had antiphospholipid syndrome, 64 (20%) had Sticky Platelet Syndrome, 38 (12%) had MTHFR mutation, 23 (7.1%) had PAI-1 polymorphism, 12 (3.7%) had Protein S deficiency, 12 (3.7%) had Factor V Leiden, 3 (1%), had AT deficiency, 3 (1%) had Heparin-Cofactor II deficiency, 3 (1%) had TPA deficiency, and 6 (2%) had Protein C deficiency. There were a total of 364 defects found in the 312 patients harboring thrombophilia; thus, several harbored two and a few harbored three separate defects.
All patients with thrombophilia were treated with preconception ASA at 81 mg. / day with the immediate post-conception addition of heparin or LMW heparin (Dalteparin). Both ASA and heparin / LMW heparin were used to term. The first 120 patients were treated with unfractionated heparin at 5,000 U every 12 hours, subcutaneously and the last 192 have been treated with Dalteparin at 5,000 U / day subcutaneously. The patients with MTHFR were also treated with folate at 5 mg. / day + pyridoxine at 50 mg. / day. All patients were carefully monitored with CBC & platelet counts, anti-Xa levels, frequent ultrasounds and physical exams. Only 2 of the thrombophilia patients suffered another miscarriage; all others had a normal term delivery. There were no pregnancy-related thromboses, no delivery complications and no episodes of post-partum thrombosis. The only bleeding consisted of 1 – 4 cm bruises at injection sites. No episodes of thrombocytopenia (HIT) were noted.
In our experience, thrombophilia is a common cause of recurrent miscariage and all patients with no anatomical, hormonal or chromosomal defect should be evaluated for thrombophilia or a bleeding disorder. The success rate of normal term delivery in these 312 patients was 94% using ASA + heparin or Dalteparin. In addition, side effects of therapy were minimal.
A Novel Mechanism of Plasmin-Induced Mitogenesis in Fibroblasts.
Authors: Mandal Samir K. , Rao L. Vijaya Mohan , Tran Thuy Tien T. , Pendurthi Usha 
Published: October 2019

Abstract
The plasminogen activator/plasmin system plays an important role, in addition to fibrinolysis, in many pathophysiological processes. Our recent studies showed that plasmin, similar to factor VIIa/tissue factor and thrombin, upregulates Cyr61, a growth factor like immediate early response gene, in fibroblasts via protease-activated receptor-mediated signaling. In preliminary studies we found that when fibroblasts were treated with VIIa (100 nM), thrombin (10 nM) or plasmin (50 nM), in concentrations that were shown to induce Cyr61 expression maximally, VIIa and thrombin increased the DNA synthesis by only about 20–30% whereas plasmin increased the DNA synthesis by 200 to 500%. The present study was carried out to investigate potential mechanism(s) by which plasmin promotes the DNA synthesis in fibroblasts and the role of protease-activated receptors and Cyr61 in plasmin-induced DNA synthesis. The plasmin-induced DNA synthesis in fibroblasts was dependent on its protease activity as the addition of active site-inhibited plasmin had no effect; and tissue factor pathway inhibitor-2 (TFPI-2) completely abrogated the plasmin-induced response. Neutralizing antibodies against PAR-1 and not PAR-2 attenuated the plasmin-induced DNA synthesis. Cyr61 antibodies completely blocked the plasmin-induced DNA synthesis suggesting that Cyr61 acts as a down-stream mediator. Surprisingly, we found no detectable increase in Cyr61 protein in plasmin-treated cell lysates whereas Cyr61 protein levels were clearly increased in thrombin-treated cells. These data suggest that plasmin might be cleaving newly synthesized Cyr61, which accumulates in ECM. Studies performed with purified recombinant Cyr61 and tumor cells that constitutively express abundant Cyr61 provided support for this notion. To test the hypothesis that plasmin release of Cyr61 from ECM plays a role in promoting DNA synthesis, we investigated the effect of conditioned media from control, plasmin-, and thrombin-treated fibroblasts in promoting DNA synthesis. The data revealed that the conditioned media of plasmin-treated cells (the protease activity in the media was inactivated) and not others significantly increased 3H-thymidine incorporation in fibroblasts. Addition of Cyr61 antibodies to the conditioned media of plasmin-treated cells attenuated the enhancing effect of the conditioned media. In summary, the data presented herein provide a novel mechanism by which plasmin promotes cell proliferation. In this mechanism, plasmin first induces the expression of Cyr61 via activation of PAR-1 mediated signaling. Next, plasmin releases/cleaves newly synthesized Cyr61 from the ECM, making it accessible to cells. The ability of plasmin to induce mitogenesis, in addition to its pericellular proteolysis, may provide a coordinated spatiotemporal effect that is required for would healing and tissue remodeling.
Interim Analysis of a Phase II Study of Denileukin Diftitox (Ontak) for Relapsed/Refractory T-Cell Non-Hodgkin’s Lymphoma.
Authors: Dang Nam H. , Pro Barbara , Hagemeister Fredrick B. , Jones Dan , Samuels Barry , Rodriguez Maria A. , Goy Andre , Romaguera Jorge E. , Walker Pamela L. , Kwak Larry , Fayad Luis 
Published: October 2019

Abstract
Denileukin diftitox (Ontak) is a fusion protein combining the enzymatically active domain of diphtheria toxin and the full-length sequence of interleukin-2 (IL-2) that targets tumor cells expressing the IL-2 receptor (IL-2R). The drug has established efficacy in cutaneous T-cell lymphoma (CTCL), and we have recently demonstrated its single-agent activity in B-cell non Hodgkin’s lymphoma (NHL) (Dang et al. Journal of Clinical Oncology. In Press). We initiated a phase II study to evaluate its efficacy in relapsed/refractory T-cell NHL, excluding CTCL. Denileukin diftitox was administered at a dose schedule of 18 μg/kg/day by IV infusion once daily for 5 days every three weeks, for up to 8 cycles. Premedications in the form of corticosteroids, antihistamines and fluids were given prior to each drug infusion to reduce the incidence and severity of acute hypersensitivity. 14 patients are currently evaluable for response. Median age was 57 (range 26–80), and mean number of previous treatments was 2.2 (range 1–4). Tumor CD25 status was determined by immunohistochemistry and/or flow cytometry, with CD25-positivity being defined as 10% or more of tumor cells expressing detectable CD25. Of the 7 patients with CD25+ T-NHL, there were 2 CR (1 case of Alk-1 negative ALCL and 1 case of PTCL), 3 PR (1 case of PTCL and 2 cases of angioimmunoblastic lymphoma), 1 SD (1 case of PTCL) and 1 PD (1 case of PTCL). Of the 7 patients with CD25− T-NHL, there were 2 PR (1 case of PTCL and 1 case of T/NK-lymphoma), 4 SD (3 cases of PTCL and 1 case of Sezary syndrome), and 1 PD (1 case of angioimmunoblastic lymphoma). Overall response rate (CR+ PR) was 50%, with 2 of 14 patients having CR (14%) and 5 of 14 patients having PR (36%). One patient with Alk-1negative ALCL still has an ongoing CR at 15+ months. Treatment was well-tolerated, with the majority of toxicity being grade 1 or 2 and transient. Denileukin diftitox appears to have activity in relapsed/refractory T-cell NHL, and is well-tolerated at the dosing schedule tested. Additional patients are being studied to further evaluate the relationship between detectable CD25 expression and tumor response to denileukin diftitox.
IL-2 Ameliorates Chronic GVHD by Inducing Antigen-Specific Regulatory T Cells.
Authors: Lohr Jens G. , Knoechel Birgit , Kahn Estelle C. , Abbas Abul K. 
Published: October 2019

Abstract
We have developed a mouse model in which a GvHD-like syndrome develops in response to a defined soluble self-antigen. This phenotype is caused after transfer of CD4+ T cells that have a single specificity and are reactive to the self-antigen into a lymphopenic host that expresses the cognate antigen. By using a clonotypic antibody we are able to identify these cells and can therefore follow their migration, kinetics and functional characteristics. At least two distinct phases can be identified by clinical picture and correlated with accumulation of T cells - an early phase, resembling acute GvHD, leading to wasting and death coinciding with rapid accumulation of T cells, and a late phase in which a stable number of T cells is maintained clinically reminiscent of chronic GvHD. We show here that a fraction of the naïve T cells that encounter the self-antigen after transfer develop into CD4+CD25+ regulatory T cells (Treg) in the periphery. This population controls T cell homeostasis, activation and severe immune pathology. The development of CD4+CD25+ Treg critically depends on IL-2 produced by the T cells. Therefore, in the absence of IL-2, T cell homeostasis cannot be maintained and massive accumulation of CD4+ T cells leads to severe inflammation of the skin. Importantly, only IL-2 that is produced by the T cells themselves, but not from peripheral tissues, leads to efficient generation of Treg and T cell homeostasis. We suggest that Treg-development is a differentiation step of T cells that encounter self-antigen in the periphery, and is essential for maintaining homeostasis even in the presence of self-recognition. Our data provide mechanistic insight into the re-establishment of homeostasis after cell transfer into a lymphopenic host and have important implications for the use and timing of therapeutic approaches targeting the IL-2 pathway.
Sphingosine 1-Phosphate (S1P) Is a Chemotactic Factor for CD34+ Progenitors and CD34+ Cell Lines Expressing the S1P1 Receptor: Possible Contribution to Progenitor Cell Trafficking.
Authors: Seitz Gabriele , Boehmler Andreas M. , Wiesner Tina , Kanz Lothar , Möhle Robert 
Published: October 2019

Abstract
Trafficking of hematopoietic stem and progenitor cells (HPC) is controlled by G protein coupled receptors (GPRs), particularly by the chemokine receptor CXCR4. However, homing of HPC does not exclusively depend on CXCR4 and its ligand SDF-1. In addition to chemokine receptors, also GPRs for non-peptide lipid mediators may be involved in HPC migration. In the present study, we demonstrate that sphingosine 1-phosphate (S1P) is chemotactic for human CD34+ HPC (optimum dose 5 μM), as measured in a modified Boyden chamber system. In CD34+ cell lines, the chemotactic response was even stronger and occurred at lower S1P concentrations (e.g., optimum dose 100 nM for Jurkat cells, 2-fold stronger migration compared to HPC). By RT-PCR, we measured mRNA expression of the 5 different S1P receptors (S1P1,2,3,4,5). All CD34+ hematopoietic cell lines analyzed (KG1, KG1a, Jurkat) expressed S1P1, a S1P receptor with known chemotactic activity, while CD34- cell lines (HL-60, THP-1) were negative for S1P1. In HL-60 cells, chemotaxis in response to S1P was even reduced compared to spontaneous migration, which might be due to the expression of S1P receptors other than S1P1 with known inhibitory effects, particularly S1P2. In mobilized peripheral blood CD34+ progenitor cells from different donors, S1P1 was consistently expressed in both CD34+CD38+ and more primitive CD34+CD38- HPC, while expression of S1P2,3,4,5 was variable. S1P induced also other typical responses of GPR-mediated signaling in CD34+ cell lines and HPC, such as polymerization of filamentous actin, as measured by flow cytometry after labeling of the cells with FITC-phalloidin. We conclude that S1P is a chemotactic factor for CD34+ HPC and CD34+ cell lines due to expression of the GPR S1P1. As megakaryocytes and platelets represent an abundant source of S1P in the bone marrow, our results suggest that S1P and its receptor S1P1 may contribute to trafficking and spatial distribution of HPC in the hematopoietic microenvironment.
EPCs Derived from Ex Vivo Expansion of Unmobilized Human Peripheral Blood Co-Express Hematopoietic and Endothelial-Specific Markers and Can Be Derived from Culture of Highly Purified Populations of CD14-Positive Monocytes.
Authors: Sharpe Emerson E. , Teleron Amylynn A. , Li Bin , Young Pampee P. 
Published: October 2019

Abstract
An increasing amount of data has suggested a more dynamic role of vasculogenesis, whereby bone marrow (BM)-derived circulating endothelial progenitor cells (EPCs) home and contribute to new blood vessel formation during tumor growth, ischemic injury, and wound healing. EPCs can be obtained by isolating hematopoietic progenitor cells from BM or cord blood. Alternatively, ex vivo expansion of unmobilized human peripheral blood (PB) can generate adherent cells, PB-EPCs, that express endothelial markers and also, upon administration, incorporate into developing neovasculature. The relative ease of obtaining unmobilized human PB has made PB-EPCs an attractive candidate with which to develop cell based therapy to treat ischemia. In parallel with clinical trials designed to understand their therapeutic potential, there is a continued effort to better characterize the PB-EPC and understand its biology. It is currently thought that EPCs are directly derived from a CD34+/lin- faction of hematopoietic stem cells (HSCs). However, in our current study, we have confirmed prior reports that ex vivo expansion of human PB generates similar numbers of EPCs as compared to plating unfractionated human BM, which contains >50-fold higher CD34+/lin- content, suggesting that PB-EPCs may not be derived from the CD34+/lin- population. We used immunofluorescence and FACs analysis to further show that PB-EPCs not only express endothelial markers such as vWF, Vascular Endothelial Growth Factor Receptor 1 and 2 (also known as flt-1 and flk-1, respectively), VE-cadherin, UEA-1 lectin, Tie-1 and Tie-2 but also hematopoietic markers such as CD45 and CD14, a marker enriched on monocytes. To test if PB-derived CD14-positive cells can give rise to PB-EPCs, we isolated them from human PB to >98% purity and plated them on fibronectin-coated coverslips. In vitro culture of CD14-positive cells generated adherent clusters of spindle shaped cells morphologically similar to EPCs. Culture of the CD14-negative fraction failed to yield any adherent cells. After ten days, the coverslips were removed and the cells were stained with various endothelial (flk-1, vWF, uptake of DiI-AcLDL, and UEA-1 lectin) and monocyte/hematopoietic (CD14 and CD45) cell markers. In analysis of these slides, the EPCs derived from the purified CD14 fraction stained positive for all six markers. These observations suggest that PB-EPCs can differentiate from cells of the monocytic lineage in vitro without the necessity of interaction from cells contained in the CD14-negative population. Further experiments will test the possibility that monocytes may be an intermediate in the differentiation of EPCs in vivo.