bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Risk of Intrauterine Growth Restriction (IGR) Is Increased by Risk Determinants of Arterial Thrombosis.
Authors: Gerhardt Andrea , Howe Nadja , Krussel Jan S. , Bender Hans G. , Scharf Rudiger E. , Zotz Rainer B. 
Published: April 2020

Abstract
Background: As previously shown, the risk of cardiovascular disease is higher after a maternal placental syndrome, especially in the presence of fetal compromise. The HPA1-b allele of the ß3 subunit of the essential platelet integrin Alphallbbeta3 is a risk factor for increased platelet thrombogenicity, leading to arterial vascular occlusion also triggered by inflammatory endothelial alterations. We performed a case-control study to assess hereditary risk factor for arterial thrombosis in addition to the known risk determinants for venous thrombosis as risk determinants for fetal IGR.
Materials and Methods: 121 women with severe fetal IGR (defined by birth weight below the 5th- percentile for gestational age and sex) and 300 normal women were evaluated. Women with other reasons of IGR (history of venous thrombosis, fetal loss, and preeclampsia, neonatale immunthrombocytopenia) were excluded. The fetuses were born alive after the 24th week of gestation.
Results: A significant risk association could be shown for the HPA-1b/1b genotype OR 3.1 (95%CI 1.2–8.8), increased levels of lipoprotein (a) (&gt;0.2g/l) OR 1.7 (95%CI 1.0–2.9), increased levels of fibrinogen (&gt;median 284mg/dl) OR 2.8 (95%CI 1.8–4.4) increased levels of FVIII:C (&gt;median 147%) OR 1.7 (95%CI 1.1–2.7), and increased levels of von-Willebrand-factor antigen (vWF-Ag) (&gt;median 129%) OR 2.7 (95%CI 1.1–2.7). The combination of risk determinants led to a further increase in risk (e.g. HPA-1b allele, increased lipoprotein (a) and vWF-Ag OR 14.0) There was no significant risk association for factor V Leiden G1691A OR 1.3 (95% CI 0.7–2.81), and prothrombin G20210A mutation. OR 2.0 (95% CI 0.6–6.5)
Conclusions: Increased platelet thrombogenicity via interaction with an inflammatory vascular process and/or via plasma levels of components of hemostasis/receptor ligands appear to be more important in women with IGR than the established risk determinants for venous thrombosis. Pharmacological control of the observed platelet thrombogenicity in patients at risk may be of clinical relevance. In consequence, it will be of importance to examine whether the critical subgroup of patients can benefit from prevention with specific antiplatelet agents.
Contributions of Phosphatidylserine Exposure on Leukemic Cells to Unregulated Procoagulant Activity in Acute Promyelocytic Leukemia.
Authors: Zhou Jin , Cao Fenglin , Lu Chengfang , Li Jinmei , Hou Jinxiao , Yu Hongjuan , Ma Ming , Liu Shuchuan , Shi Jialan 
Published: April 2020

Abstract
Acute promyelocytic leukemia (APL) causes coagulation, which can worsen following initiation of chemotherapy and is improved or corrected by the differentiating agents, all trans retinoic acid (ATRA) and arsenic trioxide (As2O3). The relationship of procoagulant activity to phosphatidylserine (PS) exposure on leukemic cells has not been clarified, although prior studies indicate that APL cells expose tissue factor (TF). Procoagulant activity of leukemic cells was measured in a modified prothrombin time in which leukemic cells replaced thromboplastin. The presence of phosphatidylserine (PS) in neutrophils and mononuclear cells (MNC) from healthy donors and APL cells from patients was investigated by flow-cytometry and confocal microscopy. The assembly of extrinsic tenase, intrinsic tenase and prothrombinase complexes on these cells was studied using enzymatic assays employing plasma or purified proteins. Lactadherin was utilized as a probe to track PS exposure and as an agent to block exposed PS. Isolated APL cells exhibited patches that stained with lactadherin, but neutrophils and MNC did not, indicating more PS exposure on APL cells than the other two cell types. Exclusion of propidium iodide by the leukemic cells indicated that PS exposure occurs in the absence of frank apoptosis. Coagulant activity increased approx 20-fold after cells were exposed to 0.1 μM daunorubicin for 24 hr with 70% of apoptosis and decreased by 85% after 24 hr treatment with 1 μM ATRA or As2O3 with partial differentiation. Inhibition of procoagulant activity by ATRA and As2O3 corresponded to decreased PS and decrease activity for all three enzyme complexes. Lactadherin, which blocks PS binding sites, inhibited Xase and prothrombin conversion by their respective APL-assembled activating complexes. APL cells support all procoagulant reactions leading to robust thrombin formation. This ability results from concomitant PS and TF exposure at the outer leaflet of cell membrane. In contrast, less PS on neutrophils and MNC do not support the coagulation.
Epac Is Involved in cAMP-Mediated Exocytosis of Weibel-Palade Bodies in Endothelial Cells through the Activation of the Small GTPase Rap1.
Authors: Voorberg Jan , Bierings Ruben , Fernandez Mar , van Mourik Jan A. 
Published: April 2020

Abstract
Vascular endothelial cells provide a dynamic interface between circulating blood and underlying tissues that is critically involved in maintaining vascular integrity and homeostasis. The endothelium provides a surface for adhesion and subsequent extravasation of leukocytes to sites of inflammation. In addition, vascular endothelial cells are involved in the regulation of vascular tone, contribute to neo-vascularisation and mediate the formation of a platelet plug in the event of vascular damage. Rapid recruitment of bio-active components from intracellular storage pools has been shown to contribute to the critical role of endothelial cells in maintaining vascular homeostasis. A significant number of haemostatic components and inflammatory mediators originate from endothelial cell-specific, cigar-shaped organelles called Weibel-Palade bodies (WPBs) that function as storage vesicles for von Willebrand factor (VWF), a multimeric adhesive glycoprotein crucial for platelet plug formation, the leukocyte receptor P-selectin and a number of bioactive compounds that include the chemoattractants IL-8 and eotaxin-3. Following stimulation with agonists that increase intracellular Ca2+ levels, such as thrombin or histamine, WPBs fuse with the plasma membrane, thereby releasing their content onto the cellular surface or into the circulation. More recently, agonists such as epinephrine and vasopressin that raise intracellular levels of cAMP have been shown to promote the release of WPBs. The physiological importance of this pathway is illustrated by the rise in VWF levels in patients with von Willebrand’s disease and mild hemophilia A following administration of the vasopressin analogue desmopressin (DDAVP) or epinephrine. We have previously shown that cAMP-mediated WPB release is dependent on protein kinase A (PKA) and involves activation of the small GTPase RalA. Here, we have investigated a possible role for another, PKA-independent cAMP-mediated signaling pathway in the regulation of WPB exocytosis, namely the exchange protein activated by cAMP (Epac) and its substrate, the small GTPase Rap1. SiRNA-mediated knockdown of the cAMP-GEF Epac1 results in decreased epinephrine-induced WPB exocytosis. Epinephrine stimulation of endothelial cells leads to activation of the small GTPase Rap1 in a PKA-independent fashion. In addition, siRNA-mediated knockdown of Epac1 completely abolished epinephrine-induced activation of Rap1 indicating that activation of Rap1 in response to epinephrine proceeds via Epac1. We subsequently addressed whether activation of Rap1 is crucial for WPB release. Downregulation of Rap1 activation through expression of Rap1GAP effectively inhibits epinephrine-induced WPB exocytosis. Taken together, these data uncover a new pathway by which endothelial cells can regulate WPB exocytosis in response to agonists that signal through cAMP.
Requirements for Retroviral Targeting of a Suicide Gene to Alloreactive Memory Stem T Cells for Adoptive Immunotherapy of Leukemia.
Authors: Bondanza Attilio , Hambach Lothar , Kaneko Shin , Mastaglio Sara , Nijmeijer Bart , Van Halteren Astrid , Ponzoni Maurilio , Aldrighetti Luca , Toma Salvatore , Radrizzani Marina , Ciceri Fabio , Bordignon Claudio , Goulmy Els , Bonini Chiara 
Published: April 2020

Abstract
In a phase I/II clinical trial investigating the prophylactic infusion of suicide gene-modified donor T cells in the context of haploidentical hemopoietic cell transplantation (haplo-HCT) for the treatment of high-risk leukemia, we observed a rapid and effective immune reconstitution. After activation with anti-CD3 antibodies, genetic modification of donor T cells was accomplished with a retroviral vector encoding for the Herpes Simplex thymidine kinase (TK). In vitro before infusion and in vivo at immune reconstitution, TK+ cells displayed an effector memory (EM) phenotype (CD45RA–CD62L−, CD28±CD27+, IL-2±IFN–γ+). The graft-versus-leukemia (GvL) effect was substantial in patients transplanted in remission, but failed to cure patients in relapse. Gene targeting with retroviral vectors is limited to memory T cells. Central memory (CM) T cells (CD45RA–CD62L+, CD28+CD27+, IL-2+IFN-γ±) share many characteristics with stem cells, namely the ability to self-renew and to differentiate into a progeny of effector cells. EM TK+ cells have a reduced alloreactivity. Recently, it has been proposed that alloreactivity may be confined to memory T cells with stem cell-features. Since alloantigens are the target not only of graft-versus-host disease (GvHD), but also of the GvL effect, crucial to the success of the strategy is the suicide gene-modification of this subset of memory T cells. We found that addition of CD28 costimulation on cell-sized beads and the use of homeostatic cytokines, such as IL-7 and IL-15, generates central memory (CM) TK+ cells. CM TK+ cells are highly alloreactive, both in vitro and in vivo in a humanized animal model of GvHD based on the grafting of human skin onto NOD/scid mice. Interestingly, CM TK+ cells express the IL7Rα, a marker associated with the stem cell-features of memory T cells. Moreover, IL7Rα expression is maintained after stimulation with alloantigens. Stimulation of CM, but not of EM TK+ cells with autologous dendritic cells pulsed with restricted peptides from the minor histocompatibility alloantigen (mHag) HA-1 or H-Y efficiently induces mHag-specific effector T cells that lyse natural ligand expressing HLA-A2+ targets. TK+ mHag-specific effector T cells also lysed mHag+HLA-A2+ leukemic cells and, when infused in conditioned NOD/scid mice harboring human leukemia, significantly delayed disease progression. Altogether, these data suggest that optimal T-cell receptor triggering and homeostatic cytokines are required for retroviral targeting of a suicide gene to alloreactive memory stem T cells and warrant their use for a safe and powerful GvL effect.
STAT3 Signaling in Donor-Derived CD4+ T-Cells Plays a Critical Role in the Induction of Acute and Chronic GVHD in Murine Models of alloBMT.
Authors: Radojcic Vedran , Pletneva Maria A. , Yen Hung-Rong , Harris Timothy J. , Pardoll Drew M. , Drake Charles G. , Luznik Leo 
Published: April 2020

Abstract
Growing evidence from several murine models suggests that STAT3 signaling in CD4+ T-cells plays a pivotal role in the pathogenesis of autoimmunity. We herein tested the hypothesis that the same pathway plays a critical role in the induction of GVHD in murine models of allogeneic BMT. To test our hypothesis and to compare the differential ability of STAT3 signaling in CD4+ T-cells to mediate GVHD in MHC-mismatched and-matched murine models of alloBMT, CD4-Cre × STAT3flox/flox (STAT3KOCD4+), or corresponding wild-type control mice were used as donors of CD4+ T-cells. In an MHC-mismatched model, groups of lethally irradiated (775 cGy) BALB/c (H-2d) mice received T-cell depleted (TCD) bone marrow (BM) from wild-type C57BL/6 (H-2b) mice alone, TCD B6 BM plus 4 × 105wild-type, or STAT3KOCD4+ T-cells, all on a C57BL/6 background. In repeated experiments we observed significantly decreased clinical signs of GVHD in animals receiving STAT3KOCD4+ in comparison to those receiving wild-type CD4+ T-cells (median GVHD score of 2.25 vs. 5.29; P &lt;.001) but no difference in lethality in chimeras monitored up to day 60 posttransplant. In an MHC-matched, minor histocompatibility mismatched setting, we conducted experiments using the B10.D2 (H-2d) BALB/c (H-2d) model in which GVHD is CD4+ T-cell-mediated and has clinicopathologic consistent with human chronic GVHD. Since the dominant clinical manifestation in this model is cutaneous sclerodermatous GVHD that is associated with in situ dermal mononuclear infiltrates, the use of BALB/c-CD45.1 (CD45.1+) mice as recipients enabled us to examine the fate of skin CD11c+ dendritic cells. After conditioning (775 cGy) recipient mice received B10.D2 (CD45.2+) TCD BM and 9.3 ×106 TCD splenocytes, repleted with 106 wild-type CD8+ and 1.9 ×106 wild-type, or STAT3KOCD4+ T-cells. We reproducibly induced all signs of chronic GVHD in chimeras receiving wild-type CD4+ T-cells, but not in chimeras injected with STAT3KOCD4+ T-cells (median GVHD score of 5.2 vs. 0.0; P &lt;.001). We found that development of cutaneous GVHD was accompanied by prominent dermal infiltration of donor-derived inflammatory monocytes and complete turnover to donor CD11c+ epidermal DC chimerism (88.4±4.3% vs. 0.9±0.6%; P &lt;.001). Interestingly, splenic CD11c+ DC (99.9±0.1% vs. 99.8±0.1%), CD4+ (99.9±0.2% vs. 99.6±0.7%) and CD8+ (99.9±0.2% vs. 99.9±0.2%) T-cell chimerism was nearly completely donor-derived and did not differ between the two sets of described chimeras. In summary, our data suggest that intact STAT3 signaling in CD4+ T-cells is required for clinical manifestations of delayed acute and/or chronic GVHD, but it does not interfere with lymphohematopoietic graft-versus host response and achievement of full donor chimerism. Further exploration of the STAT3 signaling pathway in pathophysiology of delayed acute and/or chronic GVHD is warranted.
ICSBP Haplo-Insufficiency Synergizes with SHP2-Activation for Apoptosis Resistance and Cytokine Hypersensitivity in Myeloid Progenitors.
Authors: Konieczna Iwona , Horvath Elizabeth , Wang Hao , Platanias Leonidas , Eklund Elizabeth 
Published: April 2020

Abstract
The interferon consensus sequence binding protein (ICSBP or IRF8) is an interferon regulatory transcription factor which functions as a myeloid leukemia tumor-suppressor. In mice, ICSBP-deficiency was found to induce a myeloproliferative disorder (MPD) which progressed to acute myeloid leukemia (AML) over time. This result suggested that ICSBP-deficiency was sufficient for myeloproliferation, but additional genetic lesions were required for differentiation block and therefore AML. ICSBP-deficiency has been described in human myeloid malignancies including myelodysplastic syndrome (MDS), AML and uncontrolled chronic myeloid leukemia (CML). Other previous studies determined that cytokine-induced ICSBP-tyrosine-phosphorylation was required for differentiation-stage-specific target-gene transcription during myelopoiesis. Based on these results, we hypothesized that impaired ICSBP-tyrosine-phosphorylation might exacerbate the impact of ICSBP-deficiency and induce differentiation block and progression from MPD to AML. However, leukemia-associated mutations which impaired tyrosine-phosphorylation had not been described in the gene encoding ICSBP. Therefore, we hypothesized that mutations influencing the molecular pathways involved in ICSBP-post-translational-modification might synergize with ICSBP-deficiency to accelerate progression to AML. SHP2 protein tyrosine phosphatase was previously found to dephosphorylate ICSBP in immature myeloid cells. Additional studies determined that a leukemia-associated, constitutively-active form of SHP2 de-phosphorylated ICSBP through out myelopoiesis. In the current studies, we investigated the ability of constitutive SHP2-activation to synergize with ICSBP haplo-insufficiency to induce cytokine-hypersensitivity and apoptosis-resistance. We also investigated the impact of constitutive SHP2-activation on the tumor suppressor effect of ICSBP. Both ICSBP-deficiency and constitutively active mutants of SHP2 have been described in subjects with therapy related MDS/AML. Also, activity of the bcr/abl kinase induces both SHP2 activation and ICSBP-deficiency. Therefore, our studies identified a molecular mechanism which might be involved in disease progression in these human myeloid diseases.
G-CSF Stimulates Expression of Non-Phosphorylated Stathmin (Oncoprotein, Op18) in Hematopoietic Stem Cells.
Authors: Lannert Heinrich , Able Thomas , Franz Thomas , Leicht Stefan , Wuchter Patrick , Lenze Angela , Eckstein Volker , Saffrich Rainer , Ho Anthony D. 
Published: April 2020

Abstract
Stathmin/Op18 is a cytosolic phosphoprotein which regulates the dynamics of microtubules. This regulation is important in mitosis during cell division and in the migration of cells in modification of the cytoskeleton. In this study we investigated native hematopoietic CD34+ stem cells (HSCs) from BM in comparison to mobilized peripheral blood stem cells (mPBSCs) from G-CSF stimulated healthy donors. All the cell fractions were highly enriched (&gt;99%). In comparative proteome analysis mPBSCs showed high levels of Stathmin compared to native HSCs from BM. We monitored Stathmin by fluorescent-microscopy of bone marrow smears and cytospins. We performed microarray-based gene expression profiles of the aforementioned cells focused on kinases regulating Stathmin’s activity. Western blotting and ion trap mass spectrometer (Bruker) were used to determine the differential phosphorylation. Mononuclear cells were isolated by a standard Ficoll separation method from the different blood sources. An Auto-MACS (Miltenyi) and FACS Vantage SE cell sorter (Becton Dickinson) was used to highly enrich (&gt;99%) CD34+ cells fractions.
Sample preparation: Total RNA was isolated from sorted 1 × 10e6 cells by standard methods using RNA isolation kit (Qiagen). For gene expression analysis topic-defined PIQOR™ stem cell microarrays (936 genes) were performed. Proteomics started with the determination of protein concentrations, 2D-gel-electrophoresis were described in Proteome Works System (BioRad). Sypro ruby and/or coomasie stained gels were used for protein identification after spot cutting by Q-TOF and ion trap mass spectrometer analyses.
The subcellular localization of the identified Stathmin and tubulin were performed by fluorescence and confocal microscopy of all cell fractions. The microarray gene expression correlation shows different regulated genes e.g. kinases: PAK1, MAPK, ATM1, CDKN2A, cdk, PKA, PKCB, PKCD, PKCZ. Stathmin (oncoprotein Op18) is expressed at very high levels in mPBSCs (Q-TOF), and remains non-phosphorylated (western blotting, ion trap mass spectrometry).
Our findings show that G-CSF stimulates Stathmin expression in CD34+ cells from BM and plays a key role in migration into peripheral blood. In microarray analyses we show that Stathmin phosphorylation will be regulated by different kinases in mPBSCs. But Stathmin is highly expressed in mPBSCs and is non-phosphorylated in its active form.
Identification of a Hierarchy of Multipotent Hematopoietic Progenitors in Human Cord Blood.
Authors: Majeti Ravindra , Park Christopher Y. , Weissman Irving L. 
Published: April 2020

Abstract
Mouse hematopoiesis is initiated by long-term hematopoietic stem cells (HSC) that differentiate into a series of multipotent progenitors that exhibit progressively diminished self-renewal ability. In human hematopoiesis, populations enriched for HSC have been identified, as have downstream lineage-committed progenitors, but not multipotent progenitors. Previous reports indicate that human HSC are enriched in Lin-CD34+CD38- cord blood and bone marrow, and express CD90. We demonstrate that the Lin-CD34+CD38- fraction of cord blood and bone marrow can be subdivided into three subpopulations: CD90+CD45RA-, CD90-CD45RA-, and CD90-CD45RA+. While, the function of the CD90- subpopulations is unknown, the CD90+CD45RA- subpopulation presumably contains HSC. We report here in vitro and in vivo functional studies of these three subpopulations from normal human cord blood. In vitro, CD90+CD45RA- cells formed all types of myeloid colonies in methylcellulose and were able to replate with 70% efficiency. CD90-CD45RA- cells also formed all types of myeloid colonies, but replated with only 33% efficiency. CD90-CD45RA+ cells failed to form myeloid colonies in methylcellulose. In liquid culture, CD90+CD45RA- cells gave rise to all three subpopulations; CD90-CD45RA- cells gave rise to both CD90- subpopulations, but not CD90+ cells; CD90-CD45RA+ cells gave rise to themselves only. These data establish an in vitro differentiation hierarchy from CD90+CD45RA- to CD90-CD45RA- to CD90-CD45RA+ cells among Lin-CD34+CD38- cord blood. In vivo, xenotransplantation of CD90+CD45RA- cells into NOD/SCID/IL-2R?-null newborn mice resulted in long-term multilineage engraftment with transplantation of as few as 10 purified cells. Secondary transplants from primary engrafted mice also resulted in long-term multilineage engraftment, indicating the presence of self-renewing HSC. Transplantation of CD90-CD45RA- cells also resulted in long-term multilineage engraftment; however, secondary transplants did not reliably result in long-term engraftment, indicating a reduced capacity for self-renewal. Transplantation of CD90-CD45RA+ cells did not result in any detectable human hematopoietic cells, indicating that the function of these cells is undetermined. Finally, transplantation of limiting numbers of CD90-CD45RA- cells (less than 100) resulted in multilineage human engraftment at 4 weeks, that was no longer detectable by 12 weeks. Thus, the CD90-CD45RA- subpopulation is capable of multilineage differentiation while exhibiting limited self-renewal ability. We believe this study represents the first prospective identification of a population of human multipotent progenitors, Lin-CD34+CD38-CD90-CD45RA- cord blood.