scholarly journals Mechanical Tension in Syndecan-1 is Regulated by Extracellular Mechanical Cues and Fluidic Shear Stress

2020 ◽  
Author(s):  
Victoria Le ◽  
Lei Mei ◽  
Peter L. Voyvodic ◽  
Chi Zhao ◽  
David J. Busch ◽  
...  

AbstractThe endothelium plays a central role in regulating vascular homeostasis and is key in determining the response to materials implanted in the vascular system. Endothelial cells are uniquely sensitive to biophysical cues from applied forces and their local cellular microenvironment. The glycocalyx is a layer of proteoglycans, glycoproteins and glycosaminoglycans that lines the luminal surface of the vascular endothelium, interacting directly with the components of the blood and the forces of blood flow. In this work, we examined the changes in mechanical tension of syndecan-1, a cell surface proteoglycan that is an integral part of the glycocalyx, in response to substrate stiffness and fluidic shear stress. Our studies demonstrate that syndecan-1 is mechanically responsive to extracellular mechanical cues and alters its association with cytoskeletal and adhesion-related proteins in response to substrate stiffness and physiological flow.

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Lei Mei ◽  
Victoria Le ◽  
Peter Voyvodic ◽  
Chi Zhao ◽  
David Busch ◽  
...  

Syndecan-1 (SDC1) is a transmembrane proteoglycan that mediates the shear stress-induced signaling and inflammatory phenotypes of endothelial cells. While SDC1 is known to be involved mechanically in regulating the behavior of cells, it remains unknown how SDC1 responds to extracellular mechanical cues on the molecular level. We designed a set of FRET-based SDC1 tension sensors including full-length SDC1 (SDC1TS), SDC1 with deleted ectodomain (ΔEcto), SDC1 with deleted glycosylation sites (ΔGAG) and SDC1 with a deleted cytoplasmic tail (ΔCyto). We transduced these constructs into ECs and validated the constructs by WB and confocal imaging. When cultured on glass-bottom coverslips, we found that the baseline tension in SDC1TS was significantly higher than that in Ecto and GAG mutants. When cultured on engineered substrates, we demonstrated that tension in SDC1 is modulated by micropatterned surfaces and nanotopographical cues. To study if SDC1 directly responds to substrate stiffness, transduced ECs were cultured on 0.2 kPa and 25 kPa substrates. We found that there was significantly decreased tension in the Ecto construct compared to the full-length SDC1TS construct on 0.2 kPa substrates but no significant changes were observed on the 25 kPa substrates. To examine the effect of substrate stiffness on the association of SDC1 with adhesion-related proteins, we performed IP and WB on ECs expressing HA-tagged SDC1 cultured on 0.2 kPa or 25 kPa substrates. We found there were increases in SDC1 binding to action and myosin IIb, while there were decreased in SDC1 binding to Src, PKA, and FAK in cells grown on stiff substrates versus on soft substrate. To study the effect of shear stress on the tension of SDC1, ECs expressing the constructs were cultured on static or under 12dyn/cm 2 fluidic shear stress. We found that a gradient developed in the tension of SDC1 in all four constructs. There was higher tension of SDC1 in the upstream region in comparison to the downstream region. IP and WB results showed that shear stress-induced SDC1 association with actin, Src, myosin IIb, cortactin, calmodulin, and integrin beta3. In summary, our results demonstrate that SDC1 is mechanically responsive to substrate mediated biophysical cues and shear stress.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Semra Zuhal Birol ◽  
Rana Fucucuoglu ◽  
Sertac Cadirci ◽  
Ayca Sayi-Yazgan ◽  
Levent Trabzon

AbstractAtherosclerosis is a long-term disease process of the vascular system that is characterized by the formation of atherosclerotic plaques, which are inflammatory regions on medium and large-sized arteries. There are many factors contributing to plaque formation, such as changes in shear stress levels, rupture of endothelial cells, accumulation of lipids, and recruitment of leukocytes. Shear stress is one of the main factors that regulates the homeostasis of the circulatory system; therefore, sudden and chronic changes in shear stress may cause severe pathological conditions. In this study, microfluidic channels with cavitations were designed to mimic the shape of the atherosclerotic blood vessel, where the shear stress and pressure difference depend on design of the microchannels. Changes in the inflammatory-related molecules ICAM-1 and IL-8 were investigated in THP-1 cells in response to applied shear stresses in an continuous cycling system through microfluidic channels with periodic cavitations. ICAM-1 mRNA expression and IL-8 release were analyzed by qRT-PCR and ELISA, respectively. Additionally, the adhesion behavior of sheared THP-1 cells to endothelial cells was examined by fluorescence microscopy. The results showed that 15 Pa shear stress significantly increases expression of ICAM-1 gene and IL-8 release in THP-1 cells, whereas it decreases the adhesion between THP-1 cells and endothelial cells.


Cancers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 3128
Author(s):  
Thomas R. Coughlin ◽  
Ali Sana ◽  
Kevin Voss ◽  
Abhilash Gadi ◽  
Upal Basu-Roy ◽  
...  

Osteosarcoma (OS) is an aggressive bone cancer originating in the mesenchymal lineage. Prognosis for metastatic disease is poor, with a mortality rate of approximately 40%; OS is an aggressive disease for which new treatments are needed. All bone cells are sensitive to their mechanical/physical surroundings and changes in these surroundings can affect their behavior. However, it is not well understood how OS cells specifically respond to fluid movement, or substrate stiffness—two stimuli of relevance in the tumor microenvironment. We used cells from spontaneous OS tumors in a mouse engineered to have a bone-specific knockout of pRb-1 and p53 in the osteoblast lineage. We silenced Sox2 (which regulates YAP) and tested the effect of fluid flow shear stress (FFSS) and substrate stiffness on YAP expression/activity—which was significantly reduced by loss of Sox2, but that effect was reversed by FFSS but not by substrate stiffness. Osteogenic gene expression was also reduced in the absence of Sox2 but again this was reversed by FFSS and remained largely unaffected by substrate stiffness. Thus we described the effect of two distinct stimuli on the mechanosensory and osteogenic profiles of OS cells. Taken together, these data suggest that modulation of fluid movement through, or stiffness levels within, OS tumors could represent a novel consideration in the development of new treatments to prevent their progression.


2013 ◽  
Vol 93 (5) ◽  
pp. 789-800 ◽  
Author(s):  
Katherine S. Harker ◽  
Norikiyo Ueno ◽  
Tingting Wang ◽  
Cyrille Bonhomme ◽  
Wendy Liu ◽  
...  

2021 ◽  
Author(s):  
Shigehiro Hashimoto ◽  
Hiroki Yonezawa

Abstract A cell deforms and migrates on the scaffold under mechanical stimuli in vivo. In this study, a cell with division during shear stress stimulation has been observed in vitro. Before and after division, both migration and deformation of each cell were analyzed. To make a Couette-type shear flow, the medium was sandwiched between parallel disks (the lower stationary culture-disc and the upper rotating disk) with a constant gap. The wall shear stress (1.5 Pa < τ < 2 Pa) on the surface of the lower culture plate was controlled by the rotational speed of the upper disc. Myoblasts (C2C12: mouse myoblast cell line) were used in the test. After cultivation without flow for 24 hours for adhesion of the cells to the lower disk, constant τ was applied to the cells in the incubator for 7 days. The behavior of each cell during shear was tracked by time-lapse images observed by an inverted phase contrast microscope placed in the incubator. Experimental results show that each cell tends to divide after higher activities: deformation and migration. The tendency is remarkable at the shear stress of 1.5 Pa.


Fluids ◽  
2018 ◽  
Vol 3 (4) ◽  
pp. 108 ◽  
Author(s):  
Junru Wu

Broadly speaking, acoustic streaming is generated by a nonlinear acoustic wave with a finite amplitude propagating in a viscid fluid. The fluid volume elements of molecules, d V , are forced to oscillate at the same frequency as the incident acoustic wave. Due to the nature of the nonlinearity of the acoustic wave, the second-order effect of the wave propagation produces a time-independent flow velocity (DC flow) in addition to a regular oscillatory motion (AC motion). Consequently, the fluid moves in a certain direction, which depends on the geometry of the system and its boundary conditions, as well as the parameters of the incident acoustic wave. The small scale acoustic streaming in a fluid is called “microstreaming”. When it is associated with acoustic cavitation, which refers to activities of microbubbles in a general sense, it is often called “cavitation microstreaming”. For biomedical applications, microstreaming usually takes place in a boundary layer at proximity of a solid boundary, which could be the membrane of a cell or walls of a container. To satisfy the non-slip boundary condition, the flow motion at a solid boundary should be zero. The magnitude of the DC acoustic streaming velocity, as well as the oscillatory flow velocity near the boundary, drop drastically; consequently, the acoustic streaming velocity generates a DC velocity gradient and the oscillatory flow velocity gradient produces an AC velocity gradient; they both will produce shear stress. The former is a DC shear stress and the latter is AC shear stress. It was observed the DC shear stress plays the dominant role, which may enhance the permeability of molecules passing through the cell membrane. This phenomenon is called “sonoporation”. Sonoporation has shown a great potential for the targeted delivery of DNA, drugs, and macromolecules into a cell. Acoustic streaming has also been used in fluid mixing, boundary cooling, and many other applications. The goal of this work is to give a brief review of the basic mathematical theory for acoustic microstreaming related to the aforementioned applications. The emphasis will be on its applications in biotechnology.


1967 ◽  
Vol 34 (1) ◽  
pp. 97-110 ◽  
Author(s):  
Jesse E. Sisken ◽  
Elaina Wilkes

p-Fluorophenylalanine (PFPA), an analogue of phenylalanine which may be incorporated into proteins, increases the duration of mitosis. In the present experiments, based upon quantitative analyses of time-lapse cinemicrographic films, brief treatments of cells with PFPA are shown to affect the duration of metaphase in only those cells which enter division during or shortly after treatment. The offspring of cells with prolonged metaphases also tend to have prolonged metaphases. Analyses of the kinetics of the appearance of prolonged metaphases indicate that some protein specifically associated with mitosis is synthesized primarily during a period which corresponds closely to G2. The manner in which the defect is passed on to daughter cells indicates that the protein involved is conserved and reutilized by daughter cells for their subsequent divisions. Comparable experiments performed with low concentrations of puromycin indicate that the major effect of PFPA is due to its incorporation into protein rather than its ability to inhibit protein synthesis. The fact that puromycin-induced effects can also be passed on to daughter cells is interpreted to mean that cells make only specific amounts of some mitosis-associated proteins and that if a cell "inherits" a deficiency in such protein it is not able to compensate for the deficiency.


1990 ◽  
Vol 10 (9) ◽  
pp. 4788-4794
Author(s):  
N Kabrun ◽  
N Bumstead ◽  
M J Hayman ◽  
P J Enrietto

Avian leukosis virus (ALV)-induced neoplasias are commonly found associated with integrations of proviral DNA in proximity to the myc gene. However, studies suggest that other genetic events are necessary for the complete neoplastic phenotype. A cell line (HP46) derived from an ALV-induced tumor has been analyzed and found to contain, in addition to an alteration in the myc gene, a promoter insertion in the c-rel locus. Both loci expressed large amounts of mRNA coding for their respective proteins. Several rel-related transcripts were expressed in the HP46 line, and four rel-related proteins of lower molecular weight than the wild-type p68c-rel product were detected. At least two of these transcripts contained U5 long terminal repeat sequences on the 5' end of the RNA. Structural data suggest that the messages may have evolved by an alternative splicing mechanism. This is the first example of a promoter insertion in the c-rel locus, a gene whose viral counterpart v-rel is responsible for the induction of lymphoid tumors.


2020 ◽  
Vol 11 ◽  
Author(s):  
Beibei Dai ◽  
Feng Wang ◽  
Xiang Nie ◽  
Hengzhi Du ◽  
Yanru Zhao ◽  
...  

Cardiovascular diseases are one of the prime reasons for disability and death worldwide. Diseases and conditions, such as hypoxia, pressure overload, infection, and hyperglycemia, might initiate cardiac remodeling and dysfunction by inducing hypertrophy or apoptosis in cardiomyocytes and by promoting proliferation in cardiac fibroblasts. In the vascular system, injuries decrease the endothelial nitric oxide levels and affect the phenotype of vascular smooth muscle cells. Understanding the underlying mechanisms will be helpful for the development of a precise therapeutic approach. Various microRNAs are involved in mediating multiple pathological and physiological processes in the heart. A cardiac enriched microRNA, miR-21, which is essential for cardiac homeostasis, has been demonstrated to act as a cell–cell messenger with diverse functions. This review describes the cell type–specific functions of miR-21 in different cardiovascular diseases and its prospects in clinical therapy.


2006 ◽  
Vol 18 (12) ◽  
pp. 1537-1540 ◽  
Author(s):  
D. Raz–Ben Aroush ◽  
H. D. Wagner

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