scholarly journals The Effect of Fluid Flow Shear Stress and Substrate Stiffness on Yes-Associated Protein (YAP) Activity and Osteogenesis in Murine Osteosarcoma Cells

Cancers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 3128
Author(s):  
Thomas R. Coughlin ◽  
Ali Sana ◽  
Kevin Voss ◽  
Abhilash Gadi ◽  
Upal Basu-Roy ◽  
...  
Keyword(s):  
Shear Stress ◽  
Fluid Flow ◽  
Bone Cells ◽  
Bone Cancer ◽  
Substrate Stiffness ◽  
Flow Shear ◽  
Fluid Movement ◽  
Flow Shear Stress ◽  
Fluid Flow Shear Stress ◽  
New Treatments

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.

scholarly journals Upregulated proteoglycan-related signaling pathways in fluid flow shear stress-treated podocytes

2020 ◽  
Vol 319 (2) ◽  
pp. F312-F322
Author(s):  
Tarak Srivastava ◽  
Trupti Joshi ◽  
Yuexu Jiang ◽  
Daniel P. Heruth ◽  
Mohamed H. Rezaiekhaligh ◽  
...  
Keyword(s):  
Shear Stress ◽  
Fluid Flow ◽  
Signaling Pathways ◽  
Glycogen Synthase ◽  
Prostaglandin E ◽  
Galactose Metabolism ◽  
Flow Shear ◽  
Data Set ◽  
Flow Shear Stress ◽  
Fluid Flow Shear Stress

The ultrafiltrate flow over the major processes and cell body generates fluid flow shear stress (FFSS) on podocytes. Hyperfiltration-associated increase in FFSS can lead to podocyte injury and detachment. Previously, we showed that FFSS-induced upregulation of the cyclooxygenase 2 (COX2)-PGE2-prostaglandin E receptor 2 (EP2) axis in podocytes activates Akt-glycogen synthase kinase-3β-β-catenin and MAPK/ERK signaling in response to FFSS. Integrative MultiOmics Pathway Resolution (IMPRes) is a new bioinformatic tool that enables simultaneous time-series analysis of more than two groups to identify pathways and molecular connections. In the present study, we used previously characterized COX2 [prostaglandin-endoperoxide synthase 2 ( Ptgs2)], EP2 ( Ptger2), and β1-catenin ( Ctnnb1) as “seed genes” from an array data set of four groups analyzed over a time course. The 3 seed genes shared 7 pathways and 50 genes of 14 pathways and 89 genes identified by IMPRes. A composite of signaling pathways highlighted the temporal molecular connections during mechanotransduction signaling in FFSS-treated podocytes. We investigated the “proteoglycans in cancer” and “galactose metabolism” pathways predicted by IMPRes. A custom-designed PCR array validated 60.7% of the genes predicted by IMPRes analysis, including genes for the above-named pathways. Further validation using Western blot analysis showed increased expression of phosho-Erbb2, phospho-mammalian target of rapamycin (mTOR), CD44, and hexokinase II (Hk2); decreased total Erbb2, galactose mutarotase (Galm), and β-1,4-galactosyltransferase 1 (B4galt1); and unchanged total mTOR and AKT3. These findings corroborate our previously reported results. This study demonstrates the potential of the IMPRes method to identify novel pathways. Identifying the “proteoglycans in cancer” and “galactose metabolism” pathways has generated a lead to study the significance of FFSS-induced glycocalyx remodeling and possible detachment of podocytes from the glomerular matrix.

scholarly journals Mechanotransduction signaling in podocytes from fluid flow shear stress

2018 ◽  
Vol 314 (1) ◽  
pp. F22-F34 ◽  
Author(s):  
Tarak Srivastava ◽  
Hongying Dai ◽  
Daniel P. Heruth ◽  
Uri S. Alon ◽  
Robert E. Garola ◽  
...  
Keyword(s):  
Shear Stress ◽  
Fluid Flow ◽  
Direct Effect ◽  
Exon Array ◽  
Flow Shear ◽  
Transduction Mechanisms ◽  
Flow Shear Stress ◽  
Fluid Flow Shear Stress ◽  
Biomechanical Forces ◽  
And Function

Recently, we and others have found that hyperfiltration-associated increase in biomechanical forces, namely, tensile stress and fluid flow shear stress (FFSS), can directly and distinctly alter podocyte structure and function. The ultrafiltrate flow over the major processes and cell body generates FFSS to podocytes. Our previous work suggests that the cyclooxygenase-2 (COX-2)-PGE2-PGE2 receptor 2 (EP2) axis plays an important role in mechanoperception of FFSS in podocytes. To address mechanotransduction of the perceived stimulus through EP2, cultured podocytes were exposed to FFSS (2 dyn/cm2) for 2 h. Total RNA from cells at the end of FFSS treatment, 2-h post-FFSS, and 24-h post-FFSS was used for whole exon array analysis. Differentially regulated genes ( P < 0.01) were analyzed using bioinformatics tools Enrichr and Ingenuity Pathway Analysis to predict pathways/molecules. Candidate pathways were validated using Western blot analysis and then further confirmed to be resulting from a direct effect of PGE2 on podocytes. Results show that FFSS-induced mechanotransduction as well as exogenous PGE2 activate the Akt-GSK3β-β-catenin (Ser552) and MAPK/ERK but not the cAMP-PKA signal transduction cascades. These pathways are reportedly associated with FFSS-induced and EP2-mediated signaling in other epithelial cells as well. The current regimen for treating hyperfiltration-mediated injury largely depends on targeting the renin-angiotensin-aldosterone system. The present study identifies specific transduction mechanisms and provides novel information on the direct effect of FFSS on podocytes. These results suggest that targeting EP2-mediated signaling pathways holds therapeutic significance for delaying progression of chronic kidney disease secondary to hyperfiltration.

scholarly journals Fluid flow shear stress over podocytes is increased in the solitary kidney

2013 ◽  
Vol 29 (1) ◽  
pp. 65-72 ◽  
Author(s):  
Tarak Srivastava ◽  
Gianni E. Celsi ◽  
Mukut Sharma ◽  
Hongying Dai ◽  
Ellen T. McCarthy ◽  
...  
Keyword(s):  
Shear Stress ◽  
Fluid Flow ◽  
Solitary Kidney ◽  
Flow Shear ◽  
Flow Shear Stress ◽  
Fluid Flow Shear Stress

scholarly journals Transcription Factor β-Catenin Plays a Key Role in Fluid Flow Shear Stress-Mediated Glomerular Injury in Solitary Kidney

Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1253
Author(s):  
Tarak Srivastava ◽  
Daniel P. Heruth ◽  
R. Scott Duncan ◽  
Mohammad H. Rezaiekhaligh ◽  
Robert E. Garola ◽  
...  
Keyword(s):  
Shear Stress ◽  
Transcription Factor ◽  
Fluid Flow ◽  
Immunofluorescence Microscopy ◽  
Solitary Kidney ◽  
Glomerular Injury ◽  
Flow Shear ◽  
Flow Shear Stress ◽  
Fluid Flow Shear Stress

Increased fluid flow shear stress (FFSS) in solitary kidney alters podocyte function in vivo. FFSS-treated cultured podocytes show upregulated AKT-GSK3β-β-catenin signaling. The present study was undertaken to confirm (i) the activation of β-catenin signaling in podocytes in vivo using unilaterally nephrectomized (UNX) TOPGAL mice with the β-galactosidase reporter gene for β-catenin activation, (ii) β-catenin translocation in FFSS-treated mouse podocytes, and (iii) β-catenin signaling using publicly available data from UNX mice. The UNX of TOPGAL mice resulted in glomerular hypertrophy and increased the mesangial matrix consistent with hemodynamic adaptation. Uninephrectomized TOPGAL mice showed an increased β-galactosidase expression at 4 weeks but not at 12 weeks, as assessed using immunofluorescence microscopy (p < 0.001 at 4 weeks; p = 0.16 at 12 weeks) and X-gal staining (p = 0.008 at 4 weeks; p = 0.65 at 12 weeks). Immunofluorescence microscopy showed a significant increase in phospho-β-catenin (Ser552, p = 0.005) at 4 weeks but not at 12 weeks (p = 0.935) following UNX, and the levels of phospho-β-catenin (Ser675) did not change. In vitro FFSS caused a sustained increase in the nuclear translocation of phospho-β-catenin (Ser552) but not phospho-β-catenin (Ser675) in podocytes. The bioinformatic analysis of the GEO dataset, #GSE53996, also identified β-catenin as a key upstream regulator. We conclude that transcription factor β-catenin mediates FFSS-induced podocyte (glomerular) injury in solitary kidney.

scholarly journals Prostaglandin E2 is crucial in the response of podocytes to fluid flow shear stress

2010 ◽  
Vol 4 (2) ◽  
pp. 79-90 ◽  
Author(s):  
Tarak Srivastava ◽  
Ellen T. McCarthy ◽  
Ram Sharma ◽  
Patricia A. Cudmore ◽  
Mukut Sharma ◽  
...  
Keyword(s):  
Shear Stress ◽  
Fluid Flow ◽  
Prostaglandin E2 ◽  
Flow Shear ◽  
Flow Shear Stress ◽  
Fluid Flow Shear Stress
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