33 results on '"Kushner EJ"'
Search Results
2. CD31+ T cells, endothelial function and cardiovascular risk.
- Author
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Weil BR, Kushner EJ, Diehl KJ, Greiner JJ, Stauffer BL, Desouza CA, Weil, Brian R, Kushner, Erich J, Diehl, Kyle J, Greiner, Jared J, Stauffer, Brian L, and Desouza, Christopher A
- Abstract
Deficits in endothelial cell repair mechanisms are thought to contribute to the aetiology of endothelial dysfunction and, subsequently, cardiovascular disease (CVD). CD31(+) T cells or so-called "angiogenic T cells" are a newly defined T cell subset that exhibit favourable vascular qualities and show a strong negative relation with atherosclerotic disease severity. Despite growing evidence that CD31(+) T cells are important for vascular homeostasis, it is currently unknown if CD31(+) T cell number and function are related to endothelial function and CVD risk in healthy adults. To address this question, we studied 24 healthy adult men (ages: 21-70). Endothelial function was assessed by the forearm blood flow (FBF) response to intra-arterial infusion of acetylcholine (ACh) and CVD risk was estimated by Framingham Risk Score (FRS). CD31(+) T cell number was determined by fluorescence-activated cell sorting. Magnetic-activated cell sorting was used to isolate CD31(+) T cells for Boyden chamber migration. No relation was observed between CD31(+) T cell number and FBF response to ACh or FRS. However, CD31(+) T cell migration to stromal cell-derived factor (SDF)-1α and vascular endothelial growth factor (VEGF) was positively correlated with FBF response to ACh (r = 0.43 for SDF-1α; r = 0.38 for VEGF; both P<0.05) and inversely related to FRS (r = -0.53 for SDF-1α; r = -0.48 for VEGF; both P<0.05). These findings demonstrate that CD31(+) T cell function, but not number, is associated with in vivo endothelial function and CVD risk in healthy adult men. [ABSTRACT FROM AUTHOR]
- Published
- 2011
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3. Lipidure-based micropattern fabrication for stereotyping cell geometry.
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Grespin DB, Niven TG, Babson RO, and Kushner EJ
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- Cell Adhesion, Stereotyping, Cytoskeleton
- Abstract
Cell autonomous behaviors such as migration and orchestration of cell polarity programs are required for physiological tissue formation. Micropatterns are cell-adhesive shapes that confine cell(s) to a user defined geometry. This biophysical confinement allows researchers to standardize the cell shape, and in doing so, stereotype organelle and cytoskeletal systems that can have an arbitrary organization. Thus, micropatterning can be a powerful tool in interrogation of polarity programs by enforcing a homogenous cell shape and cytoskeletal organization. A major drawback of this approach is the equipment and reagent costs associated with fabrication. Here, we provide a characterization of a compound called Lipidure (2-Methacryloyloxy ethyl phosphorylcholine) that is up to 40X less expensive than other cell repulsive coating agents. We found that Lipidure is an effective cell-repulsive agent for photolithography-based micropattern fabrication. Our results demonstrate that Lipidure is sensitive to deep UV irradiation for photolithography masking, stable in both benchtop and aqueous environments, non-toxic in prolonged culture, and effective at constraining cell geometry for quantification of cytoskeletal systems., (© 2023. The Author(s).)
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- 2023
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4. Arf6 is required for endocytosis and filamentous actin assembly during angiogenesis in vitro.
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Francis CR, Bell ML, Skripnichuk MM, and Kushner EJ
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- ADP-Ribosylation Factors genetics, ADP-Ribosylation Factors metabolism, Endothelial Cells metabolism, Endocytosis physiology, Clathrin metabolism, Intercellular Signaling Peptides and Proteins, ADP-Ribosylation Factor 6, Actins
- Abstract
Objective: Endocytosis is a process vital to angiogenesis and vascular homeostasis. In pathologies where supraphysiological growth factor signaling underlies disease etiology, such as in diabetic retinopathy and solid tumors, strategies to limit chronic growth factor signaling by way of blunting endocytic processes have been shown to have tremendous clinical value. ADP ribosylation factor 6 (Arf6) is a small GTPase that promotes the assembly of actin necessary for clathrin-mediated and clathrin-independent endocytosis. In its absence, growth factor signaling is greatly diminished, which has been shown to ameliorate pathological signaling input in diseased vasculature. However, it is less clear if there are bystander effects related to loss of Arf6 on angiogenic behaviors. Our goal was to provide an analysis of Arf6's function in angiogenic endothelium, focusing on its role in actin and endocytosis as well as sprouting morphogenesis., Methods: Primary endothelial cells were cultured in both 2D and 3D environments. Here, endothelial cells were fixed and stained for various proteins or transfected with fluorescently-tagged constructs for live-cell imaging., Results: We found that Arf6 localized to both filamentous actin and sites of endocytosis in two-dimensional culture. Loss of Arf6 distorted both apicobasal polarity and reduced the total cellular filamentous actin content, which may be the primary driver underlying gross sprouting dysmorphogenesis in its absence., Conclusions: Our findings highlight that endothelial Arf6 is a potent mediator of both actin regulation and endocytosis and is required for proper sprout formation., (© 2023 John Wiley & Sons Ltd.)
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- 2023
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5. Arf6 Regulates Endocytosis and Angiogenesis by Promoting Filamentous Actin Assembly.
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Francis CR, Bell ML, Skripnichuk MM, and Kushner EJ
- Abstract
Clathrin-mediated endocytosis (CME) is a process vital to angiogenesis as well as general vascular homeostasis. In pathologies where supraphysiological growth factor signaling underlies disease etiology, such as in diabetic retinopathy and solid tumors, strategies to limit chronic growth factor signaling by way of CME have been shown to have tremendous clinical value. ADP ribosylation factor 6 (Arf6) is a small GTPase that promotes the assembly of actin necessary for CME. In its absence, growth factor signaling is greatly diminished, which has been shown to ameliorate pathological signaling input in diseased vasculature. However, it is less clear if there are bystander effects related to loss of Arf6 on angiogenic behaviors. Our goal was to provide a analysis of Arf6’s function in angiogenic endothelium, focusing on its role in lumenogenesis as well as its relation to actin and CME. We found that Arf6 localized to both filamentous actin and sites of CME in 2-dimensional culture. Loss of Arf6 distorted both apicobasal polarity and reduced the total cellular filamentous actin content, and this may be the primary driver underlying gross dysmorphogenesis during angiogenic sprouting in its absence. Our findings highlight that endothelial Arf6 is a potent mediator of both actin regulation and CME.
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- 2023
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6. Capturing membrane trafficking events during 3D angiogenic development in vitro.
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Francis CR and Kushner EJ
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- Morphogenesis, Fibrin metabolism, Endothelial Cells metabolism, von Willebrand Factor metabolism
- Abstract
Objectives: Vesicular trafficking dictates protein localization, functional activity, and half-life, providing a critically important regulatory step in tissue development; however, there is little information detailing endothelial-specific trafficking signatures. This is due, in part, to limitations in visualizing trafficking events in endothelial tissues. Our aim in this investigation was to explore the use of a 3-dimensional (3D) in vitro sprouting model to image endothelial membrane trafficking events., Methods: Endothelial cells were challenged to grow sprouts in a fibrin bead assay. Thereafter, spouts were transfected with fluorescent proteins and stained for various cell markers. Sprouts were then imaged for trafficking events using live and fixed-cell microscopy., Results: Our results demonstrate that fibrin bead sprouts have a strong apicobasal polarity marked by apical localization of proteins moesin and podocalyxin. Comparison of trafficking mediators Rab27a and Rab35 between 3D sprouts and 2D culture showed that vesicular carriers can be imaged at high resolution, exhibiting proper membrane polarity solely in 3D sprouts. Lastly, we imaged exocytic events of von Willebrand Factor and demonstrated a distinct imaging advantage for monitoring secretion events in 3D sprouts as compared with 2D culture., Conclusions: Our results establish that the fibrin bead sprouting assay is well-suited for imaging of trafficking events during angiogenic growth., (© 2021 The Authors. Microcirculation published by John Wiley & Sons Ltd.)
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- 2022
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7. Rab35 governs apicobasal polarity through regulation of actin dynamics during sprouting angiogenesis.
- Author
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Francis CR, Kincross H, and Kushner EJ
- Subjects
- Guanine Nucleotide Exchange Factors metabolism, rab GTP-Binding Proteins genetics, rab GTP-Binding Proteins metabolism, Actins metabolism, Endothelial Cells metabolism
- Abstract
In early blood vessel development, trafficking programs, such as those using Rab GTPases, are tasked with delivering vesicular cargo with high spatiotemporal accuracy. However, the function of many Rab trafficking proteins remain ill-defined in endothelial tissue; therefore, their relevance to blood vessel development is unknown. Rab35 has been shown to play an enigmatic role in cellular behaviors which differs greatly between tissue-type and organism. Importantly, Rab35 has never been characterized for its potential contribution in sprouting angiogenesis; thus, our goal was to map Rab35's primary function in angiogenesis. Our results demonstrate that Rab35 is critical for sprout formation; in its absence, apicobasal polarity is entirely lost in vitro and in vivo. To determine mechanism, we systematically explored established Rab35 effectors and show that none are operative in endothelial cells. However, we find that Rab35 partners with DENNd1c, an evolutionarily divergent guanine exchange factor, to localize to actin. Here, Rab35 regulates actin polymerization through limiting Rac1 and RhoA activity, which is required to set up proper apicobasal polarity during sprout formation. Our findings establish that Rab35 is a potent brake of actin remodeling during blood vessel development., (© 2022. The Author(s).)
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- 2022
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8. Trafficking in blood vessel development.
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Francis CR and Kushner EJ
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- Endothelium metabolism, Protein Transport, Signal Transduction, Endothelial Cells metabolism, rab GTP-Binding Proteins metabolism
- Abstract
Blood vessels demonstrate a multitude of complex signaling programs that work in concert to produce functional vasculature networks during development. A known, but less widely studied, area of endothelial cell regulation is vesicular trafficking, also termed sorting. After moving through the Golgi apparatus, proteins are shuttled to organelles, plugged into membranes, recycled, or degraded depending on the internal and extrinsic cues. A snapshot of these protein-sorting systems can be viewed as a trafficking signature that is not only unique to endothelial tissue, but critically important for blood vessel form and function. In this review, we will cover how vesicular trafficking impacts various aspects of angiogenesis, such as sprouting, lumen formation, vessel stabilization, and secretion, emphasizing the role of Rab GTPase family members and their various effectors., (© 2022. The Author(s).)
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- 2022
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9. EHD2 modulates Dll4 endocytosis during blood vessel development.
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Webb AM, Francis CR, Judson RJ, Kincross H, Lundy KM, Westhoff DE, Meadows SM, and Kushner EJ
- Abstract
Objective: Despite the absolute requirement of Delta/Notch signaling to activate lateral inhibition during early blood vessel development, many mechanisms remain unclear about how this system is regulated. Our objective was to determine the involvement of Epsin 15 Homology Domain Containing 2 (EHD2) in delta-like ligand 4 (Dll4) endocytosis during Notch activation., Approach and Results: Using both in vivo and in vitro models, we demonstrate that EHD2 is a novel modulator of Notch activation in endothelial cells through controlling endocytosis of Dll4. In vitro, EHD2 localized to plasma membrane-bound Dll4 and caveolae. Chemical disruption of caveolae complexes resulted in EHD2 failing to organize around Dll4 as well as loss of Dll4 internalization. Reduced Dll4 internalization blunted Notch activation in endothelial cells. In vivo, EHD2 is primarily expressed in the vasculature, colocalizing with junctional marker VE-cadherin and Dll4. Knockout of EHD2 in zebrafish produced a significant increase in dysmorphic sprouts in zebrafish intersomitic vessels during development and a reduction in downstream Notch signaling., Conclusions: Overall, we demonstrate that EHD2 is necessary for Dll4 transcytosis and downstream Notch activation., (© 2021 The Authors. Microcirculation published by John Wiley & Sons Ltd.)
- Published
- 2022
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10. Correction to: Notch regulates vascular collagen IV basement membrane through modulation of lysyl hydroxylase 3 trafficking.
- Author
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Gross SJ, Webb AM, Peterlin AD, Durrant JR, Judson RJ, Raza Q, Kitajewski JK, and Kushner EJ
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- 2021
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11. Notch regulates vascular collagen IV basement membrane through modulation of lysyl hydroxylase 3 trafficking.
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Gross SJ, Webb AM, Peterlin AD, Durrant JR, Judson RJ, Raza Q, Kitajewski JK, and Kushner EJ
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- Basement Membrane, Endothelial Cells, Morphogenesis, Collagen Type IV genetics, Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase
- Abstract
Collagen type IV (Col IV) is a basement membrane protein associated with early blood vessel morphogenesis and is essential for blood vessel stability. Defects in vascular Col IV deposition are the basis of heritable disorders, such as small vessel disease, marked by cerebral hemorrhage and drastically shorten lifespan. To date, little is known about how endothelial cells regulate the intracellular transport and selective secretion of Col IV in response to angiogenic cues, leaving a void in our understanding of this critical process. Our aim was to identify trafficking pathways that regulate Col IV deposition during angiogenic blood vessel development. We have identified the GTPase Rab10 as a major regulator of Col IV vesicular trafficking during vascular development using both in vitro imaging and biochemistry as well as in vivo models. Knockdown of Rab10 reduced de novo Col IV secretion in vivo and in vitro. Mechanistically, we determined that Rab10 is an indirect mediator of Col IV secretion, partnering with atypical Rab25 to deliver the enzyme lysyl hydroxylase 3 (LH3) to Col IV-containing vesicles staged for secretion. Loss of Rab10 or Rab25 results in depletion of LH3 from Col IV-containing vesicles and rapid lysosomal degradation of Col IV. Furthermore, we demonstrate that Rab10 is Notch responsive, indicating a novel connection between permissive Notch-based vessel maturation programs and vesicle trafficking. Our results illustrate both a new trafficking-based component in the regulated secretion of Col IV and how this vesicle trafficking program interfaces with Notch signaling to fine-tune basement membrane secretion during blood vessel development., (© 2021. The Author(s).)
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- 2021
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12. Synaptotagmin-Like Protein 2a Regulates Angiogenic Lumen Formation via Weibel-Palade Body Apical Secretion of Angiopoietin-2.
- Author
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Francis CR, Claflin S, and Kushner EJ
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- Angiopoietin-2 genetics, Animals, Animals, Genetically Modified, Cells, Cultured, Humans, Membrane Proteins genetics, Receptor, TIE-2 metabolism, Signal Transduction, Weibel-Palade Bodies genetics, Zebrafish genetics, Zebrafish Proteins genetics, rab27 GTP-Binding Proteins metabolism, Angiopoietin-2 metabolism, Exocytosis, Human Umbilical Vein Endothelial Cells metabolism, Membrane Proteins metabolism, Neovascularization, Physiologic, Weibel-Palade Bodies metabolism, Zebrafish Proteins metabolism
- Abstract
[Figure: see text].
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- 2021
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13. Pericyte migration and proliferation are tightly synchronized to endothelial cell sprouting dynamics.
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Payne LB, Darden J, Suarez-Martinez AD, Zhao H, Hendricks A, Hartland C, Chong D, Kushner EJ, Murfee WL, and Chappell JC
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- Animals, Cell Proliferation, Mice, Neovascularization, Physiologic, Endothelial Cells, Pericytes
- Abstract
Pericytes are critical for microvascular stability and maintenance, among other important physiological functions, yet their involvement in vessel formation processes remains poorly understood. To gain insight into pericyte behaviors during vascular remodeling, we developed two complementary tissue explant models utilizing 'double reporter' animals with fluorescently-labeled pericytes and endothelial cells (via Ng2:DsRed and Flk-1:eGFP genes, respectively). Time-lapse confocal imaging of active vessel remodeling within adult connective tissues and embryonic skin revealed a subset of pericytes detaching and migrating away from the vessel wall. Vessel-associated pericytes displayed rapid filopodial sampling near sprouting endothelial cells that emerged from parent vessels to form nascent branches. Pericytes near angiogenic sprouts were also more migratory, initiating persistent and directional movement along newly forming vessels. Pericyte cell divisions coincided more frequently with elongating endothelial sprouts, rather than sprout initiation sites, an observation confirmed with in vivo data from the developing mouse brain. Taken together, these data suggest that (i) pericyte detachment from the vessel wall may represent an important physiological process to enhance endothelial cell plasticity during vascular remodeling, and (ii) pericyte migration and proliferation are highly synchronized with endothelial cell behaviors during the coordinated expansion of a vascular network., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)
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- 2021
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14. Excess centrosomes disrupt vascular lumenization and endothelial cell adherens junctions.
- Author
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Buglak DB, Kushner EJ, Marvin AP, Davis KL, and Bautch VL
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- Animals, Cell Polarity, Humans, Neovascularization, Physiologic, Zebrafish, Zebrafish Proteins metabolism, Adherens Junctions metabolism, Blood Vessels metabolism, Centrosome metabolism, Human Umbilical Vein Endothelial Cells metabolism
- Abstract
Proper blood vessel formation requires coordinated changes in endothelial cell polarity and rearrangement of cell-cell junctions to form a functional lumen. One important regulator of cell polarity is the centrosome, which acts as a microtubule organizing center. Excess centrosomes perturb aspects of endothelial cell polarity linked to migration, but whether centrosome number influences apical-basal polarity and cell-cell junctions is unknown. Here, we show that excess centrosomes alter the apical-basal polarity of endothelial cells in angiogenic sprouts and disrupt endothelial cell-cell adherens junctions. Endothelial cells with excess centrosomes had narrower lumens in a 3D sprouting angiogenesis model, and zebrafish intersegmental vessels had reduced perfusion following centrosome overduplication. These results indicate that endothelial cell centrosome number regulates proper lumenization downstream of effects on apical-basal polarity and cell-cell junctions. Endothelial cells with excess centrosomes are prevalent in tumor vessels, suggesting how centrosomes may contribute to tumor vessel dysfunction.
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- 2020
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15. Excess centrosomes induce p53-dependent senescence without DNA damage in endothelial cells.
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Yu Z, Ruter DL, Kushner EJ, and Bautch VL
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- Cell Culture Techniques, Cell Proliferation genetics, Cell Proliferation physiology, Humans, Tumor Suppressor Protein p53 genetics, Centrosome metabolism, DNA Damage physiology, Endothelial Cells metabolism, Neoplasms metabolism, Tumor Suppressor Protein p53 metabolism
- Abstract
Tumor blood vessels support tumor growth and progression. Centrosomes are microtubule organization centers in cells, and often up to 30% of tumor endothelial cells (ECs) acquire excess (>2) centrosomes. Although excess centrosomes can lead to aneuploidy and chromosome instability in tumor cells, how untransformed ECs respond to excess centrosomes is poorly understood. We found that the frequency of primary human ECs with excess centrosomes was quickly reduced in a p53-dependent manner. Excess centrosomes in ECs were associated with p53 phosphorylation at Ser33, increased p21 levels, and decreased cell proliferation and expression of senescence markers, but independent of DNA damage and apoptosis. Aspects of the senescence-associated phenotype were also observed in mouse ECs that were isolated from tumors with excess centrosomes. Primary ECs with excess centrosomes in vascular sprouts also had elevated Ser33 p53 phosphorylation and expressed senescence markers. Our work demonstrates that nontransformed ECs respond differently to excess centrosomes than do most tumor cells-they undergo senescence in vascular sprouts and vessels, which suggests that pathologic outcomes of centrosome overduplication depend on the transformation status of cells.-Yu, Z., Ruter, D. L., Kushner, E. J., Bautch, V. L. Excess centrosomes induce p53-dependent senescence without DNA damage in endothelial cells., (© FASEB.)
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- 2017
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16. Tumor-Derived Factors and Reduced p53 Promote Endothelial Cell Centrosome Over-Duplication.
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Yu Z, Mouillesseaux KP, Kushner EJ, and Bautch VL
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- Animals, Cells, Cultured, Centrosome drug effects, Centrosome metabolism, Down-Regulation drug effects, Down-Regulation genetics, Endothelial Cells drug effects, Endothelial Cells metabolism, Endothelial Cells ultrastructure, Female, Human Umbilical Vein Endothelial Cells, Humans, Mice, Neoplasms blood supply, Neoplasms metabolism, RNA Interference, RNA, Small Interfering pharmacology, Tumor Suppressor Protein p53 antagonists & inhibitors, Centrosome physiology, Endothelial Cells physiology, Neoplasms genetics, Neoplasms pathology, Tumor Suppressor Protein p53 genetics
- Abstract
Approximately 30% of tumor endothelial cells have over-duplicated (>2) centrosomes, which may contribute to abnormal vessel function and drug resistance. Elevated levels of vascular endothelial growth factor A induce excess centrosomes in endothelial cells, but how other features of the tumor environment affect centrosome over-duplication is not known. To test this, we treated endothelial cells with tumor-derived factors, hypoxia, or reduced p53, and assessed centrosome numbers. We found that hypoxia and elevated levels of bone morphogenetic protein 2, 6 and 7 induced excess centrosomes in endothelial cells through BMPR1A and likely via SMAD signaling. In contrast, inflammatory mediators IL-8 and lipopolysaccharide did not induce excess centrosomes. Finally, down-regulation in endothelial cells of p53, a critical regulator of DNA damage and proliferation, caused centrosome over-duplication. Our findings suggest that some tumor-derived factors and genetic changes in endothelial cells contribute to excess centrosomes in tumor endothelial cells., Competing Interests: The authors have declared that no competing interests exist.
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- 2016
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17. Notch regulates BMP responsiveness and lateral branching in vessel networks via SMAD6.
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Mouillesseaux KP, Wiley DS, Saunders LM, Wylie LA, Kushner EJ, Chong DC, Citrin KM, Barber AT, Park Y, Kim JD, Samsa LA, Kim J, Liu J, Jin SW, and Bautch VL
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- Animals, Bone Morphogenetic Proteins genetics, Cell Line, Human Umbilical Vein Endothelial Cells, Humans, Mice, Receptors, Notch genetics, Smad6 Protein genetics, Zebrafish, Bone Morphogenetic Proteins metabolism, Neovascularization, Physiologic physiology, Receptors, Notch metabolism, Smad6 Protein metabolism
- Abstract
Functional blood vessel growth depends on generation of distinct but coordinated responses from endothelial cells. Bone morphogenetic proteins (BMP), part of the TGFβ superfamily, bind receptors to induce phosphorylation and nuclear translocation of SMAD transcription factors (R-SMAD1/5/8) and regulate vessel growth. However, SMAD1/5/8 signalling results in both pro- and anti-angiogenic outputs, highlighting a poor understanding of the complexities of BMP signalling in the vasculature. Here we show that BMP6 and BMP2 ligands are pro-angiogenic in vitro and in vivo, and that lateral vessel branching requires threshold levels of R-SMAD phosphorylation. Endothelial cell responsiveness to these pro-angiogenic BMP ligands is regulated by Notch status and Notch sets responsiveness by regulating a cell-intrinsic BMP inhibitor, SMAD6, which affects BMP responses upstream of target gene expression. Thus, we reveal a paradigm for Notch-dependent regulation of angiogenesis: Notch regulates SMAD6 expression to affect BMP responsiveness of endothelial cells and new vessel branch formation.
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- 2016
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18. Excess centrosomes perturb dynamic endothelial cell repolarization during blood vessel formation.
- Author
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Kushner EJ, Ferro LS, Yu Z, and Bautch VL
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- Actin Cytoskeleton, Actins, Actomyosin, Blood Vessels metabolism, Blood Vessels physiology, Cell Culture Techniques, Cell Polarity physiology, Cytoskeleton, Dyneins, Humans, Microtubules metabolism, Tubulin, rac1 GTP-Binding Protein genetics, rac1 GTP-Binding Protein metabolism, Centrosome metabolism, Centrosome physiology, Endothelial Cells metabolism
- Abstract
Blood vessel formation requires dynamic movements of endothelial cells (ECs) within sprouts. The cytoskeleton regulates migratory polarity, and centrosomes organize the microtubule cytoskeleton. However, it is not well understood how excess centrosomes, commonly found in tumor stromal cells, affect microtubule dynamics and interphase cell polarity. Here we find that ECs dynamically repolarize during sprouting angiogenesis, and excess centrosomes block repolarization and reduce migration and sprouting. ECs with excess centrosomes initially had more centrosome-derived microtubules but, paradoxically, fewer steady-state microtubules. ECs with excess centrosomes had elevated Rac1 activity, and repolarization was rescued by blockade of Rac1 or actomyosin blockers, consistent with Rac1 activity promoting cortical retrograde actin flow and actomyosin contractility, which precludes cortical microtubule engagement necessary for dynamic repolarization. Thus normal centrosome numbers are required for dynamic repolarization and migration of sprouting ECs that contribute to blood vessel formation., (© 2016 Kushner et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).)
- Published
- 2016
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19. LGN Directs Interphase Endothelial Cell Behavior via the Microtubule Network.
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Wright CE, Kushner EJ, Du Q, and Bautch VL
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- Endothelium, Vascular metabolism, Human Umbilical Vein Endothelial Cells, Humans, Neovascularization, Physiologic, Endothelium, Vascular cytology, Interphase, Intracellular Signaling Peptides and Proteins physiology, Microtubules metabolism
- Abstract
Angiogenic sprouts require coordination of endothelial cell (EC) behaviors as they extend and branch. Microtubules influence behaviors such as cell migration and cell-cell interactions via regulated growth and shrinkage. Here we investigated the role of the mitotic polarity protein LGN in EC behaviors and sprouting angiogenesis. Surprisingly, reduced levels of LGN did not affect oriented division of EC within a sprout, but knockdown perturbed overall sprouting. At the cell level, LGN knockdown compromised cell-cell adhesion and migration. EC with reduced LGN levels also showed enhanced growth and stabilization of microtubules that correlated with perturbed migration. These results fit a model whereby LGN influences interphase microtubule dynamics in endothelial cells to regulate migration, cell adhesion, and sprout extension, and reveal a novel non-mitotic role for LGN in sprouting angiogenesis.
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- 2015
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20. Decoy receptor CXCR7 modulates adrenomedullin-mediated cardiac and lymphatic vascular development.
- Author
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Klein KR, Karpinich NO, Espenschied ST, Willcockson HH, Dunworth WP, Hoopes SL, Kushner EJ, Bautch VL, and Caron KM
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- Animals, Cell Movement, Cell Proliferation, Female, HEK293 Cells, Humans, Ligands, Male, Mice, Mice, Knockout, Muscle Cells cytology, Oligonucleotide Array Sequence Analysis, Phenotype, Receptors, CXCR genetics, Signal Transduction, Adrenomedullin metabolism, Gene Expression Regulation, Developmental, Heart embryology, Lymphatic Vessels embryology, Receptors, CXCR physiology
- Abstract
Atypical 7-transmembrane receptors, often called decoy receptors, act promiscuously as molecular sinks to regulate ligand bioavailability and consequently temper the signaling of canonical G protein-coupled receptor (GPCR) pathways. Loss of mammalian CXCR7, the most recently described decoy receptor, results in postnatal lethality due to aberrant cardiac development and myocyte hyperplasia. Here, we provide the molecular underpinning for this proliferative phenotype by demonstrating that the dosage and signaling of adrenomedullin (Adm, gene; AM, protein)-a mitogenic peptide hormone required for normal cardiovascular development-is tightly controlled by CXCR7. To this end, Cxcr7(-/-) mice exhibit gain-of-function cardiac and lymphatic vascular phenotypes that can be reversed upon genetic depletion of adrenomedullin ligand. In addition to identifying a biological ligand accountable for the phenotypes of Cxcr7(-/-) mice, these results reveal a previously underappreciated role for decoy receptors as molecular rheostats in controlling the timing and extent of GPCR-mediated cardiac and vascular development., (Copyright © 2014 Elsevier Inc. All rights reserved.)
- Published
- 2014
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21. Excess centrosomes disrupt endothelial cell migration via centrosome scattering.
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Kushner EJ, Ferro LS, Liu JY, Durrant JR, Rogers SL, Dudley AC, and Bautch VL
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- Animals, Blood Vessels pathology, Blood Vessels ultrastructure, Centrosome ultrastructure, Golgi Apparatus metabolism, Human Umbilical Vein Endothelial Cells, Humans, Interphase, Mice, Mice, Transgenic, Microtubules metabolism, Microtubules ultrastructure, Tumor Cells, Cultured, Cell Movement, Centrosome physiology, Endothelial Cells ultrastructure
- Abstract
Supernumerary centrosomes contribute to spindle defects and aneuploidy at mitosis, but the effects of excess centrosomes during interphase are poorly understood. In this paper, we show that interphase endothelial cells with even one extra centrosome exhibit a cascade of defects, resulting in disrupted cell migration and abnormal blood vessel sprouting. Endothelial cells with supernumerary centrosomes had increased centrosome scattering and reduced microtubule (MT) nucleation capacity that correlated with decreased Golgi integrity and randomized vesicle trafficking, and ablation of excess centrosomes partially rescued these parameters. Mechanistically, tumor endothelial cells with supernumerary centrosomes had less centrosome-localized γ-tubulin, and Plk1 blockade prevented MT growth, whereas overexpression rescued centrosome γ-tubulin levels and centrosome dynamics. These data support a model whereby centrosome-MT interactions during interphase are important for centrosome clustering and cell polarity and further suggest that disruption of interphase cell behavior by supernumerary centrosomes contributes to pathology independent of mitotic effects., (© 2014 Kushner et al.)
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- 2014
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22. Building blood vessels in development and disease.
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Kushner EJ and Bautch VL
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- Humans, Neovascularization, Pathologic genetics, Neovascularization, Physiologic genetics, Vascular Endothelial Growth Factor A physiology, Endothelial Cells physiology, Neoplasms blood supply, Neovascularization, Pathologic physiopathology, Neovascularization, Physiologic physiology
- Abstract
Purpose of Review: This review will examine developmental angiogenesis and tumor-related changes to endothelial cells., Recent Findings: Processes that govern developmental angiogenesis become dysfunctional in the tumor environment, leading to abnormal tumor endothelial cells and blood vessels. Recent findings suggest that tumor endothelial cells are permanently modified compared with normal counterparts., Summary: Coordination of numerous intracellular and extracellular programs promotes the formation of new blood vessels that are necessary for both development and certain diseases. Developmental angiogenesis uses canonical signaling modalities to effectively assemble endothelial cells into predictable vessel structures, and disruption of critical signaling factors has dramatic effects on blood vessel development. Solid tumors co-opt developmental cues to promote formation of tumor vessels that sustain their growth, but these angiogenic signals are not well regulated and produce endothelial cell dysfunction. Aberrant growth factor signaling contributes to phenotypic changes and acquired irreversible intracellular signaling, cytoskeletal and genetic modifications in endothelial cells of tumor vessels. Permanently altered tumor endothelial cells may represent a significant population.
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- 2013
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23. CASZ1 promotes vascular assembly and morphogenesis through the direct regulation of an EGFL7/RhoA-mediated pathway.
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Charpentier MS, Christine KS, Amin NM, Dorr KM, Kushner EJ, Bautch VL, Taylor JM, and Conlon FL
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- Animals, Blotting, Western, Calcium-Binding Proteins, Cell Adhesion, Cell Differentiation, Cell Proliferation, Cells, Cultured, Chromatin Immunoprecipitation, DNA-Binding Proteins genetics, EGF Family of Proteins, Embryo, Nonmammalian cytology, Embryonic Development, Endothelial Growth Factors genetics, Endothelium, Vascular metabolism, Extracellular Matrix Proteins genetics, Female, Fluorescent Antibody Technique, Human Umbilical Vein Endothelial Cells cytology, Human Umbilical Vein Endothelial Cells metabolism, Humans, Immunoenzyme Techniques, Molecular Sequence Data, Transcription Factors genetics, Transcription, Genetic, Xenopus Proteins genetics, Xenopus laevis growth & development, rhoA GTP-Binding Protein genetics, DNA-Binding Proteins metabolism, Embryo, Nonmammalian metabolism, Endothelial Growth Factors metabolism, Endothelium, Vascular cytology, Extracellular Matrix Proteins metabolism, Gene Expression Regulation, Morphogenesis physiology, Transcription Factors metabolism, Xenopus Proteins metabolism, Xenopus laevis metabolism, rhoA GTP-Binding Protein metabolism
- Abstract
The formation of the vascular system is essential for embryonic development and homeostasis. However, transcriptional control of this process is not fully understood. Here we report an evolutionarily conserved role for the transcription factor CASZ1 (CASTOR) in blood vessel assembly and morphogenesis. In the absence of CASZ1, Xenopus embryos fail to develop a branched and lumenized vascular system, and CASZ1-depleted human endothelial cells display dramatic alterations in adhesion, morphology, and sprouting. Mechanistically, we show that CASZ1 directly regulates Epidermal Growth Factor-Like Domain 7 (Egfl7). We further demonstrate that defects of CASZ1- or EGFL7-depleted cells are in part due to diminished RhoA expression and impaired focal adhesion localization. Moreover, these abnormal endothelial cell behaviors in CASZ1-depleted cells can be rescued by restoration of Egfl7. Collectively, these studies show that CASZ1 is required to directly regulate an EGFL7/RhoA-mediated pathway to promote vertebrate vascular development., (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Published
- 2013
- Full Text
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24. Effects of endothelin-1 on endothelial progenitor cell function.
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Diehl KJ, Weil BR, Westby CM, MacEneaney OJ, Kushner EJ, Greiner JJ, and DeSouza CA
- Subjects
- Adult, Apoptosis drug effects, Cell Movement drug effects, Humans, Stem Cells metabolism, Vascular Endothelial Growth Factor A metabolism, Young Adult, Endothelin-1 pharmacology, Endothelium, Vascular cytology, Stem Cells cytology, Stem Cells drug effects
- Abstract
Background: Circulating endothelial progenitor cells (EPCs) contribute to vascular endothelial repair. Endothelin (ET)-1 is associated with endothelial damage and atherogenesis. The experimental aim of this study was to determine, in vitro, the effects of ET-1 on the ability of EPCs to form colonies, migrate, release angiogenic growth factors and resist apoptosis., Methods: Peripheral blood samples were collected from 10 healthy adult humans. Cells with phenotypic EPC characteristics were isolated and EPC colony-forming capacity (CFU assay), migratory activity (Boyden chamber), release of angiogenic growth factors (enzyme immunoassay) and apoptosis (TUNEL assay) were determined in the absence and presence of ET-1 (100 pmol)., Results: EPC colony-forming units (42±12 vs. 39±11), migratory capacity (910±146 vs. 936±148 AU) and release of vascular endothelial growth factor (202.8±68.1 vs. 204.8±69.8 pg/mL) and granulocyte-colony stimulating factor (1294.4±378.3 vs. 1136.1±310.3 pg/mL) were not significantly affected by ET-1. EPCs treated with ET-1 demonstrated a 20% increase (p<0.05) in cellular apoptosis. The proapoptotic effect of ET-1 was abolished with ET receptor blockade as well as with apocynin, a nicotinamide adenine dinucleotide phosphate (NADPH) inhibitor., Conclusions: These results indicate that ET-1 does not affect EPC colony formation, migratory capacity or angiogenic growth factor release, but does increase EPC susceptibility to apoptosis through an NADPH-dependent mechanism. Increased EPC apoptosis may contribute to the proatherogenic effects of ET-1.
- Published
- 2012
- Full Text
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25. Aging is associated with a proapoptotic endothelial progenitor cell phenotype.
- Author
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Kushner EJ, MacEneaney OJ, Weil BR, Greiner JJ, Stauffer BL, and DeSouza CA
- Subjects
- Adult, Apoptosis Regulatory Proteins metabolism, Caspase 3 metabolism, Endothelial Cells metabolism, Hematopoietic Stem Cells metabolism, Humans, Male, Middle Aged, Phenotype, Phosphatidylinositol 3-Kinases metabolism, Proto-Oncogene Proteins c-akt metabolism, Signal Transduction physiology, Telomerase metabolism, Young Adult, Aging physiology, Apoptosis physiology, Endothelial Cells cytology, Hematopoietic Stem Cells cytology, Hematopoietic Stem Cells physiology
- Abstract
The aim of this study was to determine if aging is associated with enhanced endothelial progenitor cell (EPC) sensitivity to apoptosis. Cells with phenotypic EPC characteristics were isolated from healthy, nonobese young (age 25 ± 1 years) and older (61 ± 1 years) men. Intracellular active caspase-3 concentrations in response to staurosporine stimulation were approximately 35% higher (p < 0.05) in EPCs from older (3.15 ± 0.29 pg/ml) compared with young (2.33 ± 0.24 pg/ml) men. Protein expression of Akt, p70 S6-kinase and Bcl-2 was markedly lower (approx. 35, 75 and 60%, respectively, all p < 0.05) in EPCs from older compared with young men, whereas there were no age-related differences in either 14-3-3ε or Bax expression. Additionally, EPC telomerase activity was 57% lower (p < 0.05) in older (0.18 ± 0.11 AU) versus young (0.43 ± 0.11 AU) men. These results indicate that aging is associated with a proapoptotic EPC phenotype characterized by decreased expression of key antiapoptotic proteins associated with the PI-3-kinase signaling pathway and reduced telomerase activity. These age-related changes likely contribute, in part, to the diminished ability of EPCs to resist an apoptotic stimulus in older men. Increased susceptibility to apoptosis may contribute to the numerical and functional impairments observed in EPCs with aging., (Copyright © 2011 S. Karger AG, Basel.)
- Published
- 2011
- Full Text
- View/download PDF
26. Prehypertension and endothelial progenitor cell function.
- Author
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MacEneaney OJ, DeSouza CA, Weil BR, Kushner EJ, Van Guilder GP, Mestek ML, Greiner JJ, and Stauffer BL
- Subjects
- Apoptosis physiology, Case-Control Studies, Caspase 3 metabolism, Cell Movement physiology, Colony-Forming Units Assay, Female, Humans, Male, Middle Aged, Endothelium, Vascular cytology, Prehypertension blood, Stem Cells cytology, Stem Cells physiology
- Abstract
Prehypertension is associated with significant damage to the coronary vasculature and increased rates of adverse cardiovascular events. Circulating endothelial progenitor cells (EPCs) are critical to vascular repair and the formation of new blood vessels. We tested the hypothesis that prehypertension is associated with EPC dysfunction. Peripheral blood samples were collected from 83 middle-aged and older adults (51 male and 32 female): 40 normotensive subjects (age 53±2 years; BP 111/74±1/1 mm Hg) and 43 prehypertensive subjects (age 54±2 years; 128/77±1/1 mm Hg). EPCs were isolated from peripheral blood, and EPC colony-forming capacity (colony-forming unit (CFU) assay), migratory activity (Boyden chamber) and apoptotic susceptibility (active caspase-3 concentrations) were determined. There were no significant differences in the number of EPC CFUs (10±2 vs 9±1), EPC migration (1165±82 vs 1120±84 fluorescent units) or active intracellular caspase-3 concentrations (2.7±0.3 vs 2.3±0.2 ng ml⁻¹) between the normotensive and prehypertensive groups. When groups were stratified into low prehypertension (n=27; systolic blood pressure: 120-129 mm Hg) and high prehypertension (n=16; 130-139 mm Hg), it was found that EPCs from the high prehypertensive group produced fewer (∼65%, P<0.05) CFUs compared with the low prehypertensive (4±1 vs 12±2) and normotensive adults. In conclusion, EPC colony-forming capacity is impaired only in prehypertensive adults with systolic BP greater than 130 mm Hg. Prehypertension is not associated with migratory dysfunction or enhanced apoptosis of EPCs.
- Published
- 2011
- Full Text
- View/download PDF
27. Human aging and CD31+ T-cell number, migration, apoptotic susceptibility, and telomere length.
- Author
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Kushner EJ, Weil BR, MacEneaney OJ, Morgan RG, Mestek ML, Van Guilder GP, Diehl KJ, Stauffer BL, and DeSouza CA
- Subjects
- Adult, Age Factors, Aged, Aging genetics, Aging metabolism, Aging pathology, Cardiovascular Diseases genetics, Cardiovascular Diseases metabolism, Cardiovascular Diseases pathology, Caspase 3 metabolism, Cell Separation methods, Chemokine CXCL12 metabolism, Cytochromes c metabolism, Flow Cytometry, Humans, Lymphocyte Count, Male, Middle Aged, Risk Factors, T-Lymphocyte Subsets pathology, T-Lymphocytes pathology, Telomerase metabolism, Vascular Endothelial Growth Factor A metabolism, Young Adult, Aging immunology, Apoptosis genetics, Cardiovascular Diseases immunology, Chemotaxis, Platelet Endothelial Cell Adhesion Molecule-1 analysis, T-Lymphocyte Subsets immunology, T-Lymphocytes immunology, Telomere metabolism
- Abstract
CD31(+) T cells, or so-called "angiogenic T cells," have been shown to demonstrate vasculoprotective and neovasculogenic qualities. The influence of age on CD31(+) T-cell number and function is unclear. We tested the hypothesis that circulating CD31(+) T-cell number and migratory capacity are reduced, apoptotic susceptibility is heightened, and telomere length is shortened with advancing age in adult humans. Thirty-six healthy, sedentary men were studied: 12 young (25 ± 1 yr), 12 middle aged (46 ± 1 yr), and 12 older (64 ± 2 yr). CD31(+) T cells were isolated from peripheral blood samples by magnetic-activated cell sorting. The number of circulating CD31(+) T cells (fluorescence-activated cell sorting analysis) was lower (P < 0.01) in older (24% of CD3(+) cells) compared with middle-aged (38% of CD3(+) cells) and young (40% of CD3(+) cells) men. Migration (Boyden chamber) to both VEGF and stromal cell-derived factor-1α was markedly blunted (P < 0.05) in cells harvested from middle-aged [306.1 ± 45 and 305.6 ± 46 arbitrary units (AU), respectively] and older (231 ± 65 and 235 ± 62 AU, respectively) compared with young (525 ± 60 and 570 ± 62 AU, respectively) men. CD31(+) T cells from middle-aged and older men demonstrated greater apoptotic susceptibility, as staurosporine-stimulated intracellular caspase-3 activation was ∼ 40% higher (P < 0.05) than young. There was a progressive age-related decline in CD31(+) T-cell telomere length (young: 10,706 ± 220 bp; middle-aged: 10,179 ± 251 bp; and older: 9,324 ± 192 bp). Numerical and functional impairments in this unique T-cell subpopulation may contribute to diminished angiogenic potential and greater cardiovascular risk with advancing age.
- Published
- 2010
- Full Text
- View/download PDF
28. Endothelial progenitor cell function, apoptosis, and telomere length in overweight/obese humans.
- Author
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MacEneaney OJ, Kushner EJ, Westby CM, Cech JN, Greiner JJ, Stauffer BL, and DeSouza CA
- Subjects
- Adult, Aged, Apoptosis drug effects, Cell Movement, Chemokine CXCL12 metabolism, Cytochromes c metabolism, Enzyme Inhibitors pharmacology, Granulocyte Colony-Stimulating Factor metabolism, Humans, Middle Aged, Obesity metabolism, Plant Lectins pharmacology, Staurosporine pharmacology, Telomere, Vascular Endothelial Growth Factor A metabolism, Apoptosis physiology, Caspase 3 metabolism, Cytokines metabolism, Endothelial Cells physiology, Obesity physiopathology, Stem Cells physiology
- Abstract
Excess adiposity is associated with increased cardiovascular morbidity and mortality. Endothelial progenitor cells (EPCs) play an important role in vascular repair. We tested the hypothesis that increased adiposity is associated with EPC dysfunction, characterized by diminished capacity to release angiogenic cytokines, increased apoptotic susceptibility, reduced cell migration, and shorter telomere length. A total of 67 middle-aged and older adults (42-67 years) were studied: 25 normal weight (normal weight; BMI: 18.5-24.9 kg/m(2)) and 42 overweight/obese (overweight/obese; BMI: 25.0-34.9 kg/m(2)). Cells with phenotypic EPC characteristics were isolated from peripheral blood. EPC release of vascular endothelial growth factor (VEGF) and granulocyte colony-stimulating factor (G-CSF) was determined in the absence and presence of phytohemagglutinin (10 microg/ml). Intracellular active caspase-3 and cytochrome c concentrations were determined by immunoassay. Migratory activity of EPCs in response to VEGF (2 ng/ml) and stromal cell-derived factor-1alpha (SDF-1alpha; 10 ng/ml) was determined by Boyden chamber. Telomere length was assessed by Southern hybridization. Phytohemagglutinin-stimulated release of VEGF (90.6 +/- 7.6 vs. 127.2 +/- 11.6 pg/ml) and G-CSF (896.1 +/- 77.4 vs. 1,176.3 +/- 126.3 pg/ml) was ~25% lower (P < 0.05) in EPCs from overweight/obese vs. normal weight subjects. Staurosporine induced a ~30% greater (P < 0.05) increase in active caspase-3 in EPCs from overweight/obese (2.8 +/- 0.2 ng/ml) compared with normal weight (2.2 +/- 0.2) subjects. There were no significant differences in EPC migration to either VEGF or SDF-1alpha. Telomere length did not differ between groups. These results indicate that increased adiposity adversely affects the ability of EPCs to release proangiogenic cytokines and resist apoptosis, potentially compromising their reparative potential.
- Published
- 2010
- Full Text
- View/download PDF
29. CD31+ T cells represent a functionally distinct vascular T cell phenotype.
- Author
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Kushner EJ, MacEneaney OJ, Morgan RG, Van Engelenburg AM, Van Guilder GP, and DeSouza CA
- Subjects
- Adult, Apoptosis, CD3 Complex immunology, Cell Movement, Cytokines immunology, Humans, Male, Middle Aged, Platelet Endothelial Cell Adhesion Molecule-1 immunology, T-Lymphocytes cytology, T-Lymphocytes immunology
- Abstract
In contrast to CD3(+)/CD31(-) cells, CD3(+)/CD31(+) cells aid in endothelial repair and revascularization. There are limited data regarding the functional differences between circulating CD3(+)/CD31(+) and CD3(+)/CD31(-) cells that may contribute to their divergent cardiovascular effects. The aim of the present study was to characterize functional differences between CD3(+)/CD31(+) and CD3(+)/CD31(-) cells. To address this aim, migratory capacity, proangiogenic cytokine release and apoptotic susceptibility of CD3(+)/CD31(+) and CD3(+)/CD31(-) cells were determined. Human CD3(+)/CD31(+) and CD3(+)/CD31(-)cells from peripheral blood were isolated using magnetic-activated cell sorting. CD3(+)/CD31(+) cells demonstrated significantly higher ( approximately 60%) migratory capacity to the chemokines SDF-1alpha (655+/-99 vs. 273+/-54 AU) and VEGF (618+/-99 vs. 259+/-57 AU) vs. CD3(+)/CD31(-) cells. Release of angiogenic cytokines G-CSF, interleukin-8 and matrix metallopeptidase-9 were all approximately 100% higher (P<0.05) in CD3(+)/CD31(+) than CD3(+)/CD31(-) cells. CD3(+)/CD31(+) cells exhibited significantly higher intracellular concentrations of active caspase-3 (2.61+/-0.60 vs. 0.34+/-0.09 ng/mL) and cytochrome-c (21.8+/-1.4 vs. 13.7+/-1.0 ng/mL). In summary, CD3(+)/CD31(+) cells have greater migratory and angiogenic cytokine release capacity, but are more susceptible to apoptosis compared with CD3(+)/CD31(-) cells. Enhanced migratory capacity and angiogenic cytokine release may contribute to the vasculogenic properties of this unique T cell subpopulation., (Copyright (c) 2009 Elsevier Inc. All rights reserved.)
- Published
- 2010
- Full Text
- View/download PDF
30. Aging and endothelial progenitor cell telomere length in healthy men.
- Author
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Kushner EJ, Van Guilder GP, Maceneaney OJ, Cech JN, Stauffer BL, and DeSouza CA
- Subjects
- Adult, Age Factors, Aged, Humans, Male, Middle Aged, Young Adult, Aging physiology, Endothelial Cells physiology, Stem Cells physiology, Telomere chemistry
- Abstract
Background: Telomere length declines with age in mature endothelial cells and is thought to contribute to endothelial dysfunction and atherogenesis. Bone marrow-derived circulating endothelial progenitor cells (EPCs) are critical to vascular health as they contribute to both reendothelialization and neovascularization. We tested the hypothesis that EPC telomere length decreases with age in healthy adult humans., Methods: Peripheral blood samples were collected from 40 healthy, non-obese, sedentary men: 12 young (age 21-34 years), 12 middle-aged (43-55 years) and 16 older (57-68 years). Putative EPCs were isolated from peripheral blood mononuclear cells and telomere length was determined using genomic DNA preparation and Southern hybridization techniques., Results: EPC telomere length (base pairs) was approximately 20% (p=0.01) lower in the older (8492+523 bp) compared to the middle-aged (10,565+572 bp) and young (10,205+501 bp) men. Of note, there was no difference in EPC telomere length between the middle-aged and young men., Conclusions: These results demonstrate that EPC telomere length declines with age in healthy, sedentary men. Interestingly, telomere length was well preserved in the middle-aged compared to young men, suggesting that EPC telomere shortening occurs after the age of 55 years.
- Published
- 2009
- Full Text
- View/download PDF
31. Transcriptional regulation of beta2-microglobulin demonstrated via a novel genomic and proteomic analysis of percutaneously collected peripheral atheroma.
- Author
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Stauffer BL, Kushner EJ, Wulfman T, Zeller T, Sobus R, and Westby CM
- Subjects
- Atherosclerosis genetics, Electrophoresis, Gel, Two-Dimensional, Humans, RNA, Messenger analysis, Transcription, Genetic, beta 2-Microglobulin analysis, Atherosclerosis metabolism, Genomics, Proteomics, beta 2-Microglobulin genetics
- Published
- 2008
- Full Text
- View/download PDF
32. Gender and Endothelial Progenitor Cell Number in Middle-Aged Adults.
- Author
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Stauffer BL, Maceneaney OJ, Kushner EJ, Cech JN, Greiner JJ, Westby CM, and Desouza CA
- Abstract
BACKGROUND: Between the ages of 45 and 65 years, the prevalence of cardiovascular disease is significantly lower in women compared with men. Circulating bone marrow-derived endothelial progenitor cells (EPCs) play an important role in vascular repair. Reduced EPC number is predictive of more cardiovascular events. It is currently unknown whether there is a sex-difference in EPC number in middle-aged adults. OBJECTIVE: We tested the hypothesis that circulating EPC number is higher in middle-aged women than men. METHODS: Peripheral blood samples were collected from 58 sedentary adults, 29 men (57 ± 1 yr) and 29 women (58 ± 1 yr). Mononuclear cells were isolated and fluorescence-activated cell sorting (FACS) analysis of cells negative for CD45 was performed for those positive for CD34, and triple positive for CD34, VEGFR-2, and CD133 according to the recommendations of the International Society for Hematotherapy and Graft Engineering. RESULTS: The number of CD45(-)/CD34(+) and CD45(-)/CD34(+)/ VEGFR-2(+)/CD133(+) were not significantly different between women and men (0.055 ± 0.006% vs 0.069 ± 0.008% and 0.0013 ± 0.0003% vs 0.0018 ± 0.0004%, respectively). CONCLUSIONS: These results demonstrate no sex-difference in EPC number in middle-age adults. Therefore, it is unlikely that differences in EPC number contribute to the gender-related differences in the prevalence of cardiovascular events in this population.
- Published
- 2008
- Full Text
- View/download PDF
33. On determining the statistical parameters for pollution concentration from a truncated data set.
- Author
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Kushner EJ
- Subjects
- Mathematics, Methods, Air Pollutants analysis
- Published
- 1976
- Full Text
- View/download PDF
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