7 results on '"Wendy Stam"'
Search Results
2. Fetal and neonatal alloimmune thrombocytopenia: Current pathophysiological insights and perspectives for future diagnostics and treatment
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Wendy Stam, Gabriela Elis Wachholz, Jose Maria de Pereda, Rick Kapur, Ellen van der Schoot, and Coert Margadant
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Oncology ,Hematology - Abstract
FNAIT is a pregnancy-associated condition caused by maternal alloantibodies against paternally-inherited platelet antigens, most frequently HPA-1a on integrin β3. The clinical effects range from no symptoms to fatal intracranial hemorrhage, but underlying pathophysiological determinants are poorly understood. Accumulating evidence suggests that differential antibody-Fc-glycosylation, activation of complement/effector cells, and integrin function-blocking effects contribute to clinical outcome. Furthermore, some antibodies preferentially bind platelet integrin αIIbβ3, but others bind αvβ3 on endothelial cells and trophoblasts. Defects in endothelial cells and angiogenesis may therefore contribute to severe anti-HPA-1a associated FNAIT. Moreover, anti-HPA-1a antibodies may cause placental damage, leading to intrauterine growth restriction. We discuss current insights into diversity and actions of HPA-1a antibodies, gathered from clinical studies, in vitro studies, and mouse models. Assessment of all factors determining severity and progression of anti-HPA-1a-associated FNAIT may importantly improve risk stratification and potentially reveal novel treatment strategies, both for FNAIT and other immunohematological disorders.
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- 2022
3. MO074: SGLT2 Inhibition Promotes Intrinsic Kidney Regeneration by Cells of the Renin Lineage
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Loïs van der Pluijm, Angela Koudijs, Wendy Stam, Joris Rotmans, Kenneth W Gross, Michael Paul Pieper, Anton Jan van Zonneveld, and Roel Bijkerk
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Transplantation ,Nephrology - Abstract
BACKGROUND AND AIMS With chronic kidney disease (CKD) prevalence rapidly increasing, the need for novel therapies is rising. Sodium glucose co-transporter-2 (SGLT2) inhibitors were originally developed to treat hyperglycemia in patients with type 2 diabetes mellitus. Clinical trials with the SGLT2 inhibitor empagliflozin revealed a marked attenuation of the slope of kidney function decline, also in patients with non-diabetic CKD. The exact mechanism of this kidney sparing effect still remains to be clarified. Interestingly, cells of renin lineage (CoRL), residing in the juxtaglomerular apparatus to regulate blood pressure and fluid balance, have been demonstrated to harbor a stem cell like potential. CoRL have the ability to replenish glomerular cell number by dedifferentiating, migrating and subsequently replacing various glomerular cell types in different kidney injury mouse models. Considering that empagliflozin treatment affects renin plasma levels and electrolyte balance in patients, we hypothesized that empagliflozin could have an effect on CoRL-induced glomerular regeneration. METHOD Experiments were performed in a Ren1cre; tdTomato lineage-trace mouse strain that expresses a tomato fluorescent label in all cells derived from renin lineage. Two kidney injury mouse models were applied; bilateral ischemia reperfusion injury (bIRI) and 5/6 nephrectomy (5/6NTx). Empagliflozin (10 mg/kg) was administered daily via oral gavage for 14 days. Subsequently, mice were sacrificed and kidneys were harvested for histological analysis. RESULTS In both the bIRI and 5/6NTx model, empagliflozin intake led to an increase (>2-fold) of CoRL found in the intraglomerular regions compared with vehicle control littermates. These CoRL seemed to selectively differentiate towards different glomerular cell types per model: bIRI combined with empagliflozin administration resulted in an increase of claudin- (10-fold) and integrin-α8- (1.5-fold) tomato double positive cells, suggesting favored differentiation from CoRL to respectively a parietal epithelial or mesangial cell type. In contrast, in the empagliflozin treated 5/6NTx model, an increase (1.5-fold) in tomato-podocyn double positive cells was observed, implying more restocking of glomerular podocytes by CoRL in this model. CONCLUSION SGLT2 inhibition by empagliflozin treatment leads to increased CoRL-mediated intrinsic regeneration potential and provides the kidney with different replenished cell types in different kidney disease models. Our findings demonstrate a novel mechanism via which SGLT2 inhibition might protect against kidney injury.
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- 2022
4. Author response: Metabolic response of blood vessels to TNFα
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Anton Jan van Zonneveld, Johannes C. Schoeman, Wei Yang, Abidemi Junaid, Alireza Mashaghi, Thomas Hankemeier, and Wendy Stam
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medicine.medical_specialty ,Endocrinology ,business.industry ,Internal medicine ,medicine ,Tumor necrosis factor alpha ,business - Published
- 2020
5. Metabolic response of blood vessels to TNFα
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Alireza Mashaghi, Wei Yang, Wendy Stam, Anton Jan van Zonneveld, Thomas Hankemeier, Johannes C. Schoeman, and Abidemi Junaid
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0301 basic medicine ,QH301-705.5 ,Science ,Inflammation ,030204 cardiovascular system & hematology ,medicine.disease_cause ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,3D cell culture ,0302 clinical medicine ,Metabolomics ,Tandem Mass Spectrometry ,medicine ,Humans ,oxidative stress ,Biology (General) ,General Immunology and Microbiology ,Chemistry ,Tumor Necrosis Factor-alpha ,General Neuroscience ,General Medicine ,Phenotype ,metabolomics ,In vitro ,endothelial cells ,Cell biology ,Endothelial stem cell ,030104 developmental biology ,inflammation ,Microvessels ,Medicine ,Tumor necrosis factor alpha ,Endothelium, Vascular ,microvessels-on-a-chip ,medicine.symptom ,Oxidative stress ,Chromatography, Liquid ,Research Article ,Human - Abstract
TNFα signaling in the vascular endothelium elicits multiple inflammatory responses that drive vascular destabilization and leakage. Bioactive lipids are main drivers of these processes. In vitro mechanistic studies of bioactive lipids have been largely based on two-dimensional endothelial cell cultures that, due to lack of laminar flow and the growth of the cells on non-compliant stiff substrates, often display a pro-inflammatory phenotype. This complicates the assessment of inflammatory processes. Three-dimensional microvessels-on-a-chip models provide a unique opportunity to generate endothelial microvessels in a more physiological environment. Using an optimized targeted liquid chromatography–tandem mass spectrometry measurements of a panel of pro- and anti-inflammatory bioactive lipids, we measure the profile changes upon administration of TNFα. We demonstrate that bioactive lipid profiles can be readily detected from three-dimensional microvessels-on-a-chip and display a more dynamic, less inflammatory response to TNFα, that resembles more the human situation, compared to classical two-dimensional endothelial cell cultures., eLife digest In a range of conditions called autoimmune diseases, the immune system attacks the body rather than foreign elements. This can cause inflammation that is harmful for many organs. In particular, immune cells can produce excessive amounts of a chemical messenger called tumor necrosis factor alpha (TNFα for short), which can lead to the release of fatty molecules that damage blood vessels. This process is normally studied in blood vessels cells that are grown on a dish, without any blood movement. However, in this rigid 2D environment, the cells become ‘stressed’ and show higher levels of inflammation than in the body. This makes it difficult to assess the exact role that TNFα plays in disease. A new technology is addressing this issue by enabling scientist to culture blood vessels cells in dishes coated with gelatin. This allows the cells to organize themselves in 3D, creating tiny blood vessels in which fluids can flow. However, it was unclear whether these ‘microvessels-on-a-chip’ were better models to study the role of TNFα compared to cells grown on a plate. Here, Junaid et al. compared the levels of inflammation in blood vessels cells grown in the two environments, showing that cells are less inflamed when they are cultured in 3D. In addition, when the artificial 3D-blood vessels were exposed to TNFα, they responded more like real blood vessels than the 2D models. Finally, experiments showed that it was possible to monitor the release of fatty molecules in this environment. Together, this work suggests that microvessels-on-a-chip are better models to study how TNFα harms blood vessels. Next, systems and protocols could be develop to allow automated mass drug testing in microvessels-on-a-chip. This would help scientists to quickly screen thousands of drugs and find candidates that can protect blood vessels from TNFα.
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- 2019
6. Standardized and Scalable Assay to Study Perfused 3D Angiogenic Sprouting of iPSC-derived Endothelial Cells In Vitro
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Anton Jan van Zonneveld, Vincent van Duinen, Viola Borgdorff, Thomas Hankemeier, Arie Reijerkerk, Paul Vulto, Wendy Stam, and Valeria V. Orlova
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gradients ,assay development ,Angiogenesis ,High-throughput screening ,Cellular differentiation ,General Chemical Engineering ,Induced Pluripotent Stem Cells ,microfluidics ,Neovascularization, Physiologic ,Bioengineering ,02 engineering and technology ,high-throughput screening ,General Biochemistry, Genetics and Molecular Biology ,Neovascularization ,03 medical and health sciences ,3D cell culture ,angiogenesis ,In vivo ,medicine ,Humans ,Cells, Cultured ,030304 developmental biology ,0303 health sciences ,General Immunology and Microbiology ,Chemistry ,General Neuroscience ,Cell Differentiation ,in vitro ,021001 nanoscience & nanotechnology ,In vitro ,endothelial cells ,Cell biology ,Issue 153 ,human induced pluripotent stem cells ,Cellular Microenvironment ,Cell culture ,Microvessels ,Biological Assay ,medicine.symptom ,0210 nano-technology - Abstract
Pre-clinical drug research of vascular diseases requires in vitro models of vasculature that are amendable to high-throughput screening. However, current in vitro screening models that have sufficient throughput only have limited physiological relevance, which hinders the translation of findings from in vitro to in vivo. On the other hand, microfluidic cell culture platforms have shown unparalleled physiological relevancy in vitro, but often lack the required throughput, scalability and standardization. We demonstrate a robust platform to study angiogenesis of endothelial cells derived from human induced pluripotent stem cells (iPSC-ECs) in a physiological relevant cellular microenvironment, including perfusion and gradients. The iPSC-ECs are cultured as 40 perfused 3D microvessels against a patterned collagen-1 scaffold. Upon the application of a gradient of angiogenic factors, important hallmarks of angiogenesis can be studied, including the differentiation into tip- and stalk cell and the formation of perfusable lumen. Perfusion with fluorescent tracer dyes enables the study of permeability during and after anastomosis of the angiogenic sprouts. In conclusion, this method shows the feasibility of iPSC-derived ECs in a standardized and scalable 3D angiogenic assay that combines physiological relevant culture conditions in a platform that has the required robustness and scalability to be integrated within the drug screening infrastructure.
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- 2019
7. Abstract 157: Targeting distinct nodes of the PI3K/AKT/mTOR cascade in prostate cancer cells: impact on cell proliferation, apoptosis and pathway signaling
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Wendy Stam, Ashraf Aghai, Rute B. Marques, and Wytske M. van Weerden
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Cancer Research ,Prostate cancer ,Oncology ,Cell growth ,Apoptosis ,RPTOR ,medicine ,Cancer research ,Biology ,medicine.disease ,Protein kinase B ,PI3K/AKT/mTOR pathway ,Pathway signaling - Abstract
Inactivation of the PTEN gene is a common event in PCa, leading to activation of the PI3K/AKT/mTOR pathway and promoting PCa progression. Inhibitors targeting this pathway are currently being developed as anticancer agents. While recent clinical trials with mTORC1 inhibitors have shown limited single-agent efficacy in castration-resistant prostate cancer (CRPC), it is still not known whether targeting alternative nodes of the PI3K/AKT/mTOR cascade may improve response. The current study aimed to characterize the response of PCa cell lines to multiple inhibitors targeting the PI3K, AKT and mTOR nodes of the cascade and identify possible mechanisms of resistance. The impact of the different PI3K/AKT/mTOR inhibitors on cell proliferation and apoptosis was measured by WST-1 and ApoTox-Glo assays, respectively, in a broad panel of established PCa cell lines and primary cell cultures generated from patient-derived xenografts. PTEN-negative PCa cell lines showed strong response to PI3Kβ/δ, AKT and mTOR inhibition, whereas PTEN-positive cells were resistant to PI3Kβ/δ and showed variable sensitivity to mTOR inhibitors. MTORC1 inhibitor showed low IC50 but reached plateau at about 70-80% growth inhibition. In contrast, mTORC1/2 inhibitor could reach complete growth inhibition at ~1μM concentration, in sensitive cell lines. Furthermore, treatment with PI3K or AKT inhibitor in combination with androgen depletion induced apoptosis in various PTEN-negative models, which is consistent with previously reported synergistic effect of PI3K/AKT and androgen receptor co-targeting. The effect of the inhibitors on phosphorylation of PI3K downstream targets (AKT, PRAS40, GSK3, S6K1 and 4EBP1) was analyzed by phospho-specific western blotting. Perturbation of different nodes of the PI3K/AKT/mTOR cascade resulted in differential phosphorylation of downstream targets, but there was no evident correlation with in vitro growth inhibition. In conclusion, the current study validates PI3Kβ and AKT as alternative targets in PTEN-negative PCa, whereas PTEN-positive models showed preferential yet variable responses towards mTOR inhibitors. Citation Format: Rute B. Marques, Ashraf Aghai, Wendy Stam, Wytske M. van Weerden. Targeting distinct nodes of the PI3K/AKT/mTOR cascade in prostate cancer cells: impact on cell proliferation, apoptosis and pathway signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 157. doi:10.1158/1538-7445.AM2017-157
- Published
- 2017
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