scholarly journals Endothelial lumen signaling complexes control 3D matrix–specific tubulogenesis through interdependent Cdc42- and MT1-MMP–mediated events

Blood ◽  
2010 ◽  
Vol 115 (25) ◽  
pp. 5259-5269 ◽  
Author(s):  
Anastasia Sacharidou ◽  
Wonshill Koh ◽  
Amber N. Stratman ◽  
Anne M. Mayo ◽  
Kevin E. Fisher ◽  
...  
Keyword(s):  
Tube Formation ◽  
Dominant Negative ◽  
Collagen Matrices ◽  
Signaling Complex ◽  
Vascular Morphogenesis ◽  
Cytoplasmic Tails ◽  
3D Matrix ◽  
Dependent Signal ◽  
3D Collagen ◽  
Membrane Type

Abstract Here, we define an endothelial cell (EC) lumen signaling complex involving Cdc42, Par6b, Par3, junction adhesion molecule (Jam)–B and Jam-C, membrane type 1–matrix metalloproteinase (MT1-MMP), and integrin α2β1, which coassociate to control human EC tubulogenesis in 3D collagen matrices. Blockade of both Jam-B and Jam-C using antibodies, siRNA, or dominant-negative mutants completely interferes with lumen and tube formation resulting from a lack of Cdc42 activation, inhibition of Cdc42-GTP–dependent signal transduction, and blockade of MT1-MMP–dependent proteolysis. This process requires interdependent Cdc42 and MT1-MMP signaling, which involves Par3 binding to the Jam-B and Jam-C cytoplasmic tails, an interaction that is necessary to physically couple the components of the lumen signaling complex. MT1-MMP proteolytic activity is necessary for Cdc42 activation during EC tube formation in 3D collagen matrices but not on 2D collagen surfaces, whereas Cdc42 activation is necessary for MT1-MMP to create vascular guidance tunnels and tube networks in 3D matrices through proteolytic events. This work reveals a novel interdependent role for Cdc42-dependent signaling and MT1-MMP–dependent proteolysis, a process that occurs selectively in 3D collagen matrices and that requires EC lumen signaling complexes, to control human EC tubulogenesis during vascular morphogenesis.

scholarly journals Human platelet lysate improves human cord blood derived ECFC survival and vasculogenesis in three dimensional (3D) collagen matrices

2015 ◽  
Vol 101 ◽  
pp. 72-81 ◽  
Author(s):  
Hyojin Kim ◽  
Nutan Prasain ◽  
Sasidhar Vemula ◽  
Michael J. Ferkowicz ◽  
Momoko Yoshimoto ◽  
...  
Keyword(s):  
Cord Blood ◽  
Human Platelet ◽  
Three Dimensional ◽  
Platelet Lysate ◽  
Human Cord Blood ◽  
Collagen Matrices ◽  
Human Platelet Lysate ◽  
3D Collagen

scholarly journals Alpha/Beta Interferons Potentiate Virus-Induced Apoptosis through Activation of the FADD/Caspase-8 Death Signaling Pathway

2000 ◽  
Vol 74 (3) ◽  
pp. 1513-1523 ◽  
Author(s):  
Siddharth Balachandran ◽  
P. Christopher Roberts ◽  
Todd Kipperman ◽  
Kapil N. Bhalla ◽  
Richard W. Compans ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Virus Infection ◽  
Sindbis Virus ◽  
Influenza Virus Infection ◽  
Dominant Negative ◽  
Caspase 8 ◽  
Signaling Complex ◽  
Dependent Protein ◽  
Induced Apoptosis ◽  
Dominant Negative Variant

ABSTRACT Interferon (IFN) mediates its antiviral effects by inducing a number of responsive genes, including the double-stranded RNA (dsRNA)-dependent protein kinase, PKR. Here we report that inducible overexpression of functional PKR in murine fibroblasts sensitized cells to apoptosis induced by influenza virus, while in contrast, cells expressing a dominant-negative variant of PKR were completely resistant. We determined that the mechanism of influenza virus-induced apoptosis involved death signaling through FADD/caspase-8 activation, while other viruses such as vesicular stomatitis virus (VSV) and Sindbis virus (SNV) did not significantly provoke PKR-mediated apoptosis but did induce cytolysis of fibroblasts via activation of caspase-9. Significantly, treatment with IFN-α/β greatly sensitized the fibroblasts to FADD-dependent apoptosis in response to dsRNA treatment or influenza virus infection but completely protected the cells against VSV and SNV replication in the absence of any cellular destruction. The mechanism by which IFN increases the cells' susceptibility to lysis by dsRNA or certain virus infection is by priming cells to FADD-dependent apoptosis, possibly by regulating the activity of the death-induced signaling complex (DISC). Conversely, IFN is also able to prevent the replication of viruses such as VSV that avoid triggering FADD-mediated DISC activity, by noncytopathic mechanisms, thus preventing destruction of the cell.

Abstract 614: Shear Stress-Induced Glycolytic Metabolites Promote Vascular Repair

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Kyung In Baek ◽  
Rongsong Li ◽  
David Liem ◽  
Tyler Beebe ◽  
Jianguo Ma ◽  
...  
Keyword(s):  
Shear Stress ◽  
Rabbit Model ◽  
Zealand White Rabbit ◽  
Tube Formation ◽  
Dominant Negative ◽  
Vegf Receptor ◽  
Vascular Repair ◽  
Aortic Endothelial Cells ◽  
Enhanced Tube ◽  
Vegfr Inhibitor

Introduction: Hemodynamic shear stress is intimately linked with transcriptomic and epigenomic changes to maintain endothelial homeostasis. Metabolomics studies have led to emergent metabolic biomarkers and therapeutic targets. Whether shear stress modulates metabolomic pathway to promote vascular repair remains to be investigated. Hypothesis: We hypothesized that shear stress regulates VEGF receptor-PKCε-PFKFB3 signaling-mediated glycolytic metabolites to promote vascular repair. Method and Results: Both pulsatile (PSS: 23 ± 8 dyn·cm -2 at 1 Hz) and oscillatory shear stress (OSS: 0.1 ± 3 dyn·cm -2 at 1 Hz) up-regulated PKCε expressions and the activity (* P < 0.05, n =3), whereas silencing VEGFR2 with siRNA, or treating with VEGFR inhibitor, Cediranib, attenuated shear stress-mediated PKCε expression in human aortic endothelial cells(HAEC). Constitutively active (CA)-PKCε adenovirus infection enhanced tube formation assessed by Matrigel as well as significantly increased PFKFB3 expressions promoting glycolysis, whereas the dominant negative(DN) PKCε resulted in opposite effects. Co-localization of PKCε and PFKFB3 expression was demonstrated in the endothelium of aortic arch and thoracic aorta in a New Zealand White rabbit model. In the zebrafish tail amputation model, reduction of shear stress via GATA-1a morpholino oligonucleotide(MO) injection and inhibition of PKCε expression via PKCε MO impaired vascular repair between the dorsal aorta and the dorsal longitudinal anastomotic vessel at 3 days post amputation(dpa). PKCε mRNA rescued GATA-1a MO-mediated impairment of vascular repair (* P < 0.01, n =20, ** P < 0.05, n =5). Metabolomic analysis in HAEC applied to PSS and OSS revealed modulation of a number of metabolites including increased glycolytic metabolite dihydroxyacetone, which was blocked by PKCε siRNA. Treatment with dihydroxyacetone rescued PKCε-impaired vascular repair. Conclusion: In conclusion, shear stress-mediated VEGFR-PKCε-PFKFB3 signaling increased glycolytic metabolites to mediate vascular repair.

scholarly journals LRP-1 Promotes Colon Cancer Cell Proliferation in 3D Collagen Matrices by Mediating DDR1 Endocytosis

2020 ◽  
Vol 8 ◽  
Author(s):  
Cao Cuong Le ◽  
Amar Bennasroune ◽  
Guillaume Collin ◽  
Cathy Hachet ◽  
Véronique Lehrter ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Cell Proliferation ◽  
Cancer Cell ◽  
Colon Cancer Cell ◽  
Cancer Cell Proliferation ◽  
Collagen Matrices ◽  
3D Collagen

scholarly journals Matrix Remodeling and Hyaluronan Production by Myofibroblasts and Cancer-Associated Fibroblasts in 3D Collagen Matrices

Gels ◽  
2020 ◽  
Vol 6 (4) ◽  
pp. 33
Author(s):  
Jiranuwat Sapudom ◽  
Claudia Damaris Müller ◽  
Khiet-Tam Nguyen ◽  
Steve Martin ◽  
Ulf Anderegg ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Tumor Microenvironment ◽  
Average Molecular Weight ◽  
Cell Types ◽  
Dermal Fibroblasts ◽  
Matrix Remodeling ◽  
Cancer Associated Fibroblasts ◽  
Collagen Matrices ◽  
3D Collagen ◽  
Tgf Β1

The tumor microenvironment is a key modulator in cancer progression and has become a novel target in cancer therapy. An increase in hyaluronan (HA) accumulation and metabolism can be found in advancing tumor progression and are often associated with aggressive malignancy, drug resistance and poor prognosis. Wound-healing related myofibroblasts or activated cancer-associated fibroblasts (CAF) are assumed to be the major sources of HA. Both cell types are capable to synthesize new matrix components as well as reorganize the extracellular matrix. However, to which extent myofibroblasts and CAF perform these actions are still unclear. In this work, we investigated the matrix remodeling and HA production potential in normal human dermal fibroblasts (NHFB) and CAF in the absence and presence of transforming growth factor beta -1 (TGF-β1), with TGF-β1 being a major factor of regulating fibroblast differentiation. Three-dimensional (3D) collagen matrix was utilized to mimic the extracellular matrix of the tumor microenvironment. We found that CAF appeared to response insensitively towards TGF-β1 in terms of cell proliferation and matrix remodeling when compared to NHFB. In regards of HA production, we found that both cell types were capable to produce matrix bound HA, rather than a soluble counterpart, in response to TGF-β1. However, activated CAF demonstrated higher HA production when compared to myofibroblasts. The average molecular weight of produced HA was found in the range of 480 kDa for both cells. By analyzing gene expression of HA metabolizing enzymes, namely hyaluronan synthase (HAS1-3) and hyaluronidase (HYAL1-3) isoforms, we found expression of specific isoforms in dependence of TGF-β1 present in both cells. In addition, HAS2 and HYAL1 are highly expressed in CAF, which might contribute to a higher production and degradation of HA in CAF matrix. Overall, our results suggested a distinct behavior of NHFB and CAF in 3D collagen matrices in the presence of TGF-β1 in terms of matrix remodeling and HA production pointing to a specific impact on tumor modulation.

Local, Three-Dimensional Strain Measurements Within Largely Deformed Extracellular Matrix Constructs

2004 ◽  
Vol 126 (6) ◽  
pp. 699-708 ◽  
Author(s):  
Blayne A. Roeder ◽  
Klod Kokini ◽  
J. Paul Robinson ◽  
Sherry L. Voytik-Harbin
Keyword(s):  
Extracellular Matrix ◽  
Local Level ◽  
Three Dimensional ◽  
Collagen Matrices ◽  
Mechanical Loads ◽  
Strain Measurements ◽  
3D Collagen ◽  
Vitro Model

The ability to create extracellular matrix (ECM) constructs that are mechanically and biochemically similar to those found in vivo and to understand how their properties affect cellular responses will drive the next generation of tissue engineering strategies. To date, many mechanisms by which cells biochemically communicate with the ECM are known. However, the mechanisms by which mechanical information is transmitted between cells and their ECM remain to be elucidated. “Self-assembled” collagen matrices provide an in vitro-model system to study the mechanical behavior of ECM. To begin to understand how the ECM and the cells interact mechanically, the three-dimensional (3D) mechanical properties of the ECM must be quantified at the micro-(local) level in addition to information measured at the macro-(global) level. Here we describe an incremental digital volume correlation (IDVC) algorithm to quantify large (>0.05) 3D mechanical strains in the microstructure of 3D collagen matrices in response to applied mechanical loads. Strain measurements from the IDVC algorithm rely on 3D confocal images acquired from collagen matrices under applied mechanical loads. The accuracy and the precision of the IDVC algorithm was verified by comparing both image volumes collected in succession when no deformation was applied to the ECM (zero strain) and image volumes to which simulated deformations were applied in both 1D and 3D (simulated strains). Results indicate that the IDVC algorithm can accurately and precisely determine the 3D strain state inside largely deformed collagen ECMs. Finally, the usefulness of the algorithm was demonstrated by measuring the microlevel 3D strain response of a collagen ECM loaded in tension.

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