The Experts below are selected from a list of 13620 Experts worldwide ranked by ideXlab platform
John M. Tarbell - One of the best experts on this subject based on the ideXlab platform.
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the cancer cell glycocalyx proteoglycan glypican 1 mediates Interstitial Flow mechanotransduction to enhance cell migration and metastasis
Biorheology, 2019Co-Authors: Heriberto Moran, Henry Qazi, Limary M Cancel, Mariya Mayer, Lance L Munn, John M. TarbellAbstract:Background Previous studies have demonstrated that the glycosaminoglycans (GAGs) heparan sulfate (HS) and hyaluronic acid (HA) are mechanosensors for Interstitial Flow on cancer cells. The proteins that link the GAGs to the cancer cell for mechanotransduction, however, are not known. Objective To assess whether the HS proteoglycan core proteins, Glypican-1 and Syndecan-1, or the HA receptor, CD44, provides the mechanical linkage to the cell. Methods The highly metastatic renal carcinoma cell line (SN12L1) and its companion low metastatic cell line (SN12C) were analyzed by Western blot, siRNA, and a 3-dimensional Interstitial Flow migration assay. Results There was significant elevation of Glypican-1 protein expression in the SN12L1 cells relative to the SN12C cells while there were no significant differences in Syndecan-1 or CD44. Knock down of Glypican-1 by siRNA completely blocked Flow induced migration in SN12L1 cells. MAPK inhibitors also blocked Flow induced migration in SN12L1 cells. Conclusions Glypican-1 provides the mechanical linkage from HS (the Flow sensor) to the SN12L1 cell where mechanotransduction leading to the enhancement of migration (metastasis) occurs. MAPKs downstream of Glypican-1 propagate the signal. The HS, Glypican-1, MAPK signaling axis suggests opportunities for pharmaceutical intervention.
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cancer cell glycocalyx mediates mechanotransduction and Flow regulated invasion
Integrative Biology, 2013Co-Authors: Henry Qazi, Zhong Dong Shi, Lance L Munn, Rocio Palomino, John M. TarbellAbstract:Mammalian cells are covered by a surface proteoglycan (glycocalyx) layer, and it is known that blood vessel-lining endothelial cells use the glycocalyx to sense and transduce the shearing forces of blood Flow into intracellular signals. Tumor cells in vivo are exposed to forces from Interstitial fluid Flow that may affect metastatic potential but are not reproduced by most in vitro cell motility assays. We hypothesized that glycocalyx-mediated mechanotransduction of Interstitial Flow shear stress is an un-recognized factor that can significantly enhance metastatic cell motility and play a role in augmentation of invasion. Involvement of MMP levels, cell adhesion molecules (CD44, α3 integrin), and glycocalyx components (heparan sulfate and hyaluronan) was investigated in a cell/collagen gel suspension model designed to mimic the Interstitial Flow microenvironment. Physiological levels of Flow upregulated MMP levels and enhanced the motility of metastatic cells. Blocking the Flow-enhanced expression of MMP activity or adhesion molecules (CD44 and integrins) resulted in blocking the Flow-enhanced migratory activity. The presence of a glycocalyx-like layer was verified around tumor cells, and the degradation of this layer by hyaluronidase and heparinase blocked the Flow-regulated invasion. This study shows for the first time that Interstitial Flow enhancement of metastatic cell motility can be mediated by the cell surface glycocalyx – a potential target for therapeutics.
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Effect of the glycocalyx layer on transmission of Interstitial Flow shear stress to embedded cells
Biomechanics and Modeling in Mechanobiology, 2013Co-Authors: John M. Tarbell, Zhong Dong ShiAbstract:In this paper, a simple theoretical model is developed to describe the transmission of force from Interstitial fluid Flow to the surface of a cell covered by a proteoglycan / glycoprotein layer (glycocalyx) and embedded in an extracellular matrix. Brinkman equations are used to describe Flow through the extracellular matrix and glycocalyx layers and the solid mechanical stress developed in the glycocalyx by the fluid Flow loading is determined. Using reasonable values for the Darcy permeability of extracellular matrix and glycocalyx layers and Interstitial Flow velocity, we are able to estimate the fluid and solid shear stresses imposed on the surface of embedded vascular, cartilage and tumor cells in vivo and in vitro. The principal finding is that the surface solid stress is typically one to two orders of magnitude larger than the surface fluid stress. This indicates that Interstitial Flow shear stress can be sensed by the cell surface glycocalyx, supporting numerous recent observations that Interstitial Flow can induce mechanotransduction in embedded cells. This study may contribute to understanding of Interstitial Flow-related mechanobiology in embryogenesis, tumorigenesis, tissue physiology and diseases and has implications in tissue engineering.
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Heparan Sulfate Proteoglycans Mediate Interstitial Flow Mechanotransduction Regulating MMP-13 Expression and Cell Motility via FAK-ERK in 3D Collagen
PloS one, 2011Co-Authors: Zhong Dong Shi, Hui Wang, John M. TarbellAbstract:Background Interstitial Flow directly affects cells that reside in tissues and regulates tissue physiology and pathology by modulating important cellular processes including proliferation, differentiation, and migration. However, the structures that cells utilize to sense Interstitial Flow in a 3-dimensional (3D) environment have not yet been elucidated. Previously, we have shown that Interstitial Flow upregulates matrix metalloproteinase (MMP) expression in rat vascular smooth muscle cells (SMCs) and fibroblasts/myofibroblasts via activation of an ERK1/2-c-Jun pathway, which in turn promotes cell migration in collagen. Herein, we focused on uncovering the Flow-induced mechanotransduction mechanism in 3D.
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Shear stress modulation of smooth muscle cell marker genes in 2-D and 3-D depends on mechanotransduction by heparan sulfate proteoglycans and ERK1/2.
PloS one, 2010Co-Authors: Zhong Dong Shi, Giya Abraham, John M. TarbellAbstract:Background During vascular injury, vascular smooth muscle cells (SMCs) and fibroblasts/myofibroblasts (FBs/MFBs) are exposed to altered luminal blood Flow or transmural Interstitial Flow. We investigate the effects of these two types of fluid Flows on the phenotypes of SMCs and MFBs and the underlying mechanotransduction mechanisms. Methodology/principal findings Exposure to 8 dyn/cm(2) laminar Flow shear stress (2-dimensional, 2-D) for 15 h significantly reduced expression of alpha-smooth muscle actin (alpha-SMA), smooth muscle protein 22 (SM22), SM myosin heavy chain (SM-MHC), smoothelin, and calponin. Cells suspended in collagen gels were exposed to Interstitial Flow (1 cmH(2)O, approximately 0.05 dyn/cm(2), 3-D), and after 6 h of exposure, expression of SM-MHC, smoothelin, and calponin were significantly reduced, while expression of alpha-SMA and SM22 were markedly enhanced. PD98059 (an ERK1/2 inhibitor) and heparinase III (an enzyme to cleave heparan sulfate) significantly blocked the effects of laminar Flow on gene expression, and also reversed the effects of Interstitial Flow on SM-MHC, smoothelin, and calponin, but enhanced Interstitial Flow-induced expression of alpha-SMA and SM22. SMCs and MFBs have similar responses to fluid Flow. Silencing ERK1/2 completely blocked the effects of both laminar Flow and Interstitial Flow on SMC marker gene expression. Western blotting showed that both types of Flows induced ERK1/2 activation that was inhibited by disruption of heparan sulfate proteoglycans (HSPGs). Conclusions/significance The results suggest that HSPG-mediated ERK1/2 activation is an important mechanotransduction pathway modulating SMC marker gene expression when SMCs and MFBs are exposed to Flow. Fluid Flow may be involved in vascular remodeling and lesion formation by affecting phenotypes of vascular wall cells. This study has implications in understanding the Flow-related mechanobiology in vascular lesion formation, tumor cell invasion, and stem cell differentiation.
M Swartz - One of the best experts on this subject based on the ideXlab platform.
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Abstract 2614: A three-dimensionalin vitrobioreactor for analysis of the tumor microenvironment: investigation of Interstitial Flow and the mechanism of metastasis.
Tumor Biology, 2013Co-Authors: Kathryn Hockemeyer, M Swartz, Tammy Sobolik, Alexander Terekhov, William H. Hofmeister, John P. Wikswo, Chris Janetopoulos, Ann RichmondAbstract:As recent decades have seen extensive resources dedicated to exploring tumor metastasis, discoveries have exposed a need for understanding the intricacies of the tumor microenvironment (TME) in an ex vivo model. Much of the heterogeneity of the disease stems from the complexity of interactions between the cells that comprise the TME, placing its investigation at the heart of developing effective treatment. A number of findings implicate chemokines and their receptors, expressed early in cancer progression, in promoting tumor metastasis. These findings identify several cell types significantly involved in secreting and responding to chemokines, such as tumor cells, fibroblasts, and endothelial cells, among others. Chemokine expression depends not only on the cell types of the TME but also on the conditions in the surrounding matrix, especially Interstitial Flow. A strong link has been identified between Interstitial Flow and tumor metastasis. This Flow, a result of rapid lymphangiogenesis, correlates with the expression of a number of chemokines responsible for tumor invasion and metastasis. Current limitations for studying the human TME have stalled progress since mouse models cannot fully engage all the relevant human cell types. Thus, the emergence of complex intercellular relationships within the TME generates a need for heterotypic three-dimensional in vitro models to cover the gaps between animal models and human studies. The need for 3D in vitro models is amplified by the exposed importance of Interstitial Flow. To mimic and examine the breast cancer tumor, we have designed a bioreactor, machined through femtosecond laser-etching technology that allows for co-culture of the numerous cell types that comprise the TME. The design incorporates a semicircular chamber connected to a channel 200 μm in width by a thin porous filter of laser-etched glass. The device is coated with poly-L-lysine and Collagen I to improve cell adhesion. To recreate the TME, the semicircular chamber is loaded with cancer-associated fibroblasts and tumor cells (MCF-7 and MDA-MB-231 cell lines) in a ratio of 3:1 in 3D reconstituted basement membrane with 1.5 mg/mL Collagen I (rat-tail) and 10% Matrigel. This 3D cell culture is grown for 5 days to allow spheroid formation. Human microvascular endothelial cells are cultured to form a confluent monolayer in the channel that is then perfused with medium, of which a higher quantity is aspirated from the outlet to generate Interstitial Flow through the 3D cell culture. Cancer-associated leukocytes can be infused through the channel to better mimic the TME. This unique and innovative design will allow in vitro visualization of cell migration and expression, which can shed light on the mechanisms of metastasis. Experiments can then be performed to elucidate the specific relationships linking Interstitial Flow and metastasis. Citation Format: Kathryn Hockemeyer, Tammy Sobolik, Alexander Terekhov, Melody Swartz, William Hofmeister, John Wikswo, Chris Janetopoulos, Ann Richmond. A three-dimensional in vitro bioreactor for analysis of the tumor microenvironment: investigation of Interstitial Flow and the mechanism of metastasis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2614. doi:10.1158/1538-7445.AM2013-2614
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Interstitial Flow in a 3D Microenvironment Increases Glioma Invasion by a CXCR4-Dependent Mechanism
Cancer research, 2012Co-Authors: Jennifer M. Munson, Ravi V. Bellamkonda, M SwartzAbstract:Brain tumor invasion leads to recurrence and resistance to treatment. Glioma cells invade in distinct patterns, possibly determined by microenvironmental cues including chemokines, structural heterogeneity, and fluid Flow. We hypothesized that Flow originating from pressure differentials between the brain and tumor is active in glioma invasion. Using in vitro models, we show that Interstitial Flow promotes cell invasion in multiple glioma cell lines. Flow effects were CXCR4-dependent, because they were abrogated by CXCR4 inhibition. Furthermore, CXCR4 was activated in response to Flow, which could be responsible for enhanced cell motility. Flow was seen to enhance cell polarization in the Flow direction, and this Flow-induced polarization could be blocked by CXCR4 inhibition or CXCL12 oversaturation in the matrix. Furthermore, using live imaging techniques in a three-dimensional Flow chamber, there were more cells migrating and more cells migrating in the direction of Flow. This study shows that Interstitial Flow is an active regulator of glioma invasion. The new mechanisms of glioma invasion that we identify here—namely, Interstitial Flow-enhanced motility, activation of CXCR4, and CXCL12-driven autologous chemotaxis—are significant in therapy to prevent or treat brain cancer invasion. Current treatment strategies can lead to edema and altered Flow in the brain, and one popular experimental treatment in clinical trials, convection enhanced delivery, involves enhancement of Flow in and around the tumor. A better understanding of how Interstitial Flow at the tumor margin can alter chemokine distributions, cell motility, and directed invasion offers a better understanding of treatment failure. Cancer Res; 73(5); 1536–46. ©2012 AACR .
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Migration dynamics of breast cancer cells in a tunable 3D Interstitial Flow chamber
Integrative biology : quantitative biosciences from nano to macro, 2011Co-Authors: Ulrike Haessler, Jeremy C.m. Teo, Didier Foretay, Philippe Renaud, M SwartzAbstract:The migration of cells such as leukocytes, tumor cells, and fibroblasts through 3D matrices is critical for regulating homeostasis and immunity and for driving pathogenesis. Interstitial Flow through the extracellular matrix, which can substantially increase during inflammation and in the tumor microenvironment, can influence cell migration in multiple ways. Leukocytes and tumor cells are heterogeneous in their migration responses to Flow, yet most 3D migration studies use endpoint measurements representing average characteristics. Here we present a robust new microfluidic device for 3D culture with live imaging under well-controlled Flow conditions, along with a comparison of analytical methods for describing the migration behavior of heterogeneous cell populations. We then use the model to provide new insight on how Interstitial Flow affects MDA-MB-231 breast cancer cell invasion, phenomena that are not seen from averaged or endpoint measurements. Specifically, we find that Interstitial Flow increases the percentage of cells that become migratory, and increases migrational speed in about 20% of the cells. It also increases the migrational persistence of a subpopulation (5–10% of cells) in the positive or negative Flow direction. Cells that migrated upstream moved faster but with less directedness, whereas cells that migrated in the direction of Flow moved at slower speeds but with higher directedness. These findings demonstrate how fluid Flow in the tumor microenvironment can enhance tumor cell invasion by directing a subpopulation of tumor cells in the Flow direction; i.e., towards the draining lymphatic vessels, a major route of metastasis.
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Tumor Cell Invasion Is Promoted by Interstitial Flow-Induced Matrix Priming by Stromal Fibroblasts
Cancer research, 2011Co-Authors: Adrian C Shieh, Hallie A. Rozansky, Boris Hinz, M SwartzAbstract:Interstitial Flow emanates from tumors into the microenvironment where it promotes tumor cell invasion. Fibroblasts are key constituents of the tumor stroma that modulate the mechanical environment by matrix remodeling and contraction. Here, we explore how Interstitial fluid Flow affects fibroblast–tumor cell interactions. Using a 3-dimensional invasion assay and MDA-MB-435S cells cocultured with dermal fibroblasts in a collagen matrix, we showed a synergistic enhancement of tumor cell invasion by fibroblasts in the presence of Interstitial Flow. Interstitial Flow also drove transforming growth factor (TGF)-β1 and collagenase-dependent fibroblast migration, consistent with previously described mechanisms in which Flow promotes invasion through autologous chemotaxis and increased motility. Concurrently, migrating fibroblasts enhanced tumor cell invasion by matrix priming via Rho-mediated contraction. We propose a model in which Interstitial Flow promotes fibroblast migration through increased TGF-β1 activation and collagen degradation, positioning fibroblasts to locally reorganize collagen fibers via Rho-dependent contractility, in turn enhancing tumor cell invasion via mechanotactic cues. This represents a novel mechanism in which Interstitial Flow causes fibroblast-mediated stromal remodeling that facilitates tumor invasion. Cancer Res; 71(3); 790–800. ©2011 AACR.
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A multichamber fluidic device for 3D cultures under Interstitial Flow with live imaging: Development, characterization, and applications
Biotechnology and bioengineering, 2010Co-Authors: Carmen Bonvin, Adrian C Shieh, Jan Overney, J. Brandon Dixon, M SwartzAbstract:Interstitial Flow is an important biophysical cue that can affect capillary morphogenesis, tumor cell migration, and fibroblast remodeling of the extracellular matrix, among others. Current models that incorporate Interstitial Flow and that are suitable for live imaging lack the ability to perform multiple simultaneous experiments, for example, to compare effects of growth factors, extracellular matrix composition, etc. We present a nine-chamber radial Flow device that allows simultaneous 3D fluidic experiments for relatively long-term culture with live imaging capabilities. Flow velocity profiles were characterized by fluorescence recovery after photobleaching (FRAP) for Flow uniformity and estimating the hydraulic conductivity. We demonstrate lymphatic and blood capillary morphogenesis in fibrin gels over 10 days, comparing Flow with static conditions as well as the effects of an engineered variant of VEGF that binds fibrin via Factor XIII. We also demonstrate the culture of contractile fibroblasts and co-cultures with tumor cells for modeling the tumor microenvironment. Therefore, this device is useful for studies of capillary morphogenesis, cell migration, contractile cells like fibroblasts, and multicellular cultures, all under Interstitial Flow.
Zhong Dong Shi - One of the best experts on this subject based on the ideXlab platform.
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cancer cell glycocalyx mediates mechanotransduction and Flow regulated invasion
Integrative Biology, 2013Co-Authors: Henry Qazi, Zhong Dong Shi, Lance L Munn, Rocio Palomino, John M. TarbellAbstract:Mammalian cells are covered by a surface proteoglycan (glycocalyx) layer, and it is known that blood vessel-lining endothelial cells use the glycocalyx to sense and transduce the shearing forces of blood Flow into intracellular signals. Tumor cells in vivo are exposed to forces from Interstitial fluid Flow that may affect metastatic potential but are not reproduced by most in vitro cell motility assays. We hypothesized that glycocalyx-mediated mechanotransduction of Interstitial Flow shear stress is an un-recognized factor that can significantly enhance metastatic cell motility and play a role in augmentation of invasion. Involvement of MMP levels, cell adhesion molecules (CD44, α3 integrin), and glycocalyx components (heparan sulfate and hyaluronan) was investigated in a cell/collagen gel suspension model designed to mimic the Interstitial Flow microenvironment. Physiological levels of Flow upregulated MMP levels and enhanced the motility of metastatic cells. Blocking the Flow-enhanced expression of MMP activity or adhesion molecules (CD44 and integrins) resulted in blocking the Flow-enhanced migratory activity. The presence of a glycocalyx-like layer was verified around tumor cells, and the degradation of this layer by hyaluronidase and heparinase blocked the Flow-regulated invasion. This study shows for the first time that Interstitial Flow enhancement of metastatic cell motility can be mediated by the cell surface glycocalyx – a potential target for therapeutics.
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Effect of the glycocalyx layer on transmission of Interstitial Flow shear stress to embedded cells
Biomechanics and Modeling in Mechanobiology, 2013Co-Authors: John M. Tarbell, Zhong Dong ShiAbstract:In this paper, a simple theoretical model is developed to describe the transmission of force from Interstitial fluid Flow to the surface of a cell covered by a proteoglycan / glycoprotein layer (glycocalyx) and embedded in an extracellular matrix. Brinkman equations are used to describe Flow through the extracellular matrix and glycocalyx layers and the solid mechanical stress developed in the glycocalyx by the fluid Flow loading is determined. Using reasonable values for the Darcy permeability of extracellular matrix and glycocalyx layers and Interstitial Flow velocity, we are able to estimate the fluid and solid shear stresses imposed on the surface of embedded vascular, cartilage and tumor cells in vivo and in vitro. The principal finding is that the surface solid stress is typically one to two orders of magnitude larger than the surface fluid stress. This indicates that Interstitial Flow shear stress can be sensed by the cell surface glycocalyx, supporting numerous recent observations that Interstitial Flow can induce mechanotransduction in embedded cells. This study may contribute to understanding of Interstitial Flow-related mechanobiology in embryogenesis, tumorigenesis, tissue physiology and diseases and has implications in tissue engineering.
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Heparan Sulfate Proteoglycans Mediate Interstitial Flow Mechanotransduction Regulating MMP-13 Expression and Cell Motility via FAK-ERK in 3D Collagen
PloS one, 2011Co-Authors: Zhong Dong Shi, Hui Wang, John M. TarbellAbstract:Background Interstitial Flow directly affects cells that reside in tissues and regulates tissue physiology and pathology by modulating important cellular processes including proliferation, differentiation, and migration. However, the structures that cells utilize to sense Interstitial Flow in a 3-dimensional (3D) environment have not yet been elucidated. Previously, we have shown that Interstitial Flow upregulates matrix metalloproteinase (MMP) expression in rat vascular smooth muscle cells (SMCs) and fibroblasts/myofibroblasts via activation of an ERK1/2-c-Jun pathway, which in turn promotes cell migration in collagen. Herein, we focused on uncovering the Flow-induced mechanotransduction mechanism in 3D.
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Shear stress modulation of smooth muscle cell marker genes in 2-D and 3-D depends on mechanotransduction by heparan sulfate proteoglycans and ERK1/2.
PloS one, 2010Co-Authors: Zhong Dong Shi, Giya Abraham, John M. TarbellAbstract:Background During vascular injury, vascular smooth muscle cells (SMCs) and fibroblasts/myofibroblasts (FBs/MFBs) are exposed to altered luminal blood Flow or transmural Interstitial Flow. We investigate the effects of these two types of fluid Flows on the phenotypes of SMCs and MFBs and the underlying mechanotransduction mechanisms. Methodology/principal findings Exposure to 8 dyn/cm(2) laminar Flow shear stress (2-dimensional, 2-D) for 15 h significantly reduced expression of alpha-smooth muscle actin (alpha-SMA), smooth muscle protein 22 (SM22), SM myosin heavy chain (SM-MHC), smoothelin, and calponin. Cells suspended in collagen gels were exposed to Interstitial Flow (1 cmH(2)O, approximately 0.05 dyn/cm(2), 3-D), and after 6 h of exposure, expression of SM-MHC, smoothelin, and calponin were significantly reduced, while expression of alpha-SMA and SM22 were markedly enhanced. PD98059 (an ERK1/2 inhibitor) and heparinase III (an enzyme to cleave heparan sulfate) significantly blocked the effects of laminar Flow on gene expression, and also reversed the effects of Interstitial Flow on SM-MHC, smoothelin, and calponin, but enhanced Interstitial Flow-induced expression of alpha-SMA and SM22. SMCs and MFBs have similar responses to fluid Flow. Silencing ERK1/2 completely blocked the effects of both laminar Flow and Interstitial Flow on SMC marker gene expression. Western blotting showed that both types of Flows induced ERK1/2 activation that was inhibited by disruption of heparan sulfate proteoglycans (HSPGs). Conclusions/significance The results suggest that HSPG-mediated ERK1/2 activation is an important mechanotransduction pathway modulating SMC marker gene expression when SMCs and MFBs are exposed to Flow. Fluid Flow may be involved in vascular remodeling and lesion formation by affecting phenotypes of vascular wall cells. This study has implications in understanding the Flow-related mechanobiology in vascular lesion formation, tumor cell invasion, and stem cell differentiation.
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Shear stress modulation of smooth muscle cell marker genes in 2-D and 3-D depends on mechanotransduction by heparan sulfate proteoglycans and ERK1/2.
PloS one, 2010Co-Authors: Zhong Dong Shi, Giya Abraham, John M. TarbellAbstract:During vascular injury, vascular smooth muscle cells (SMCs) and fibroblasts/myofibroblasts (FBs/MFBs) are exposed to altered luminal blood Flow or transmural Interstitial Flow. We investigate the effects of these two types of fluid Flows on the phenotypes of SMCs and MFBs and the underlying mechanotransduction mechanisms. Exposure to 8 dyn/cm(2) laminar Flow shear stress (2-dimensional, 2-D) for 15 h significantly reduced expression of alpha-smooth muscle actin (alpha-SMA), smooth muscle protein 22 (SM22), SM myosin heavy chain (SM-MHC), smoothelin, and calponin. Cells suspended in collagen gels were exposed to Interstitial Flow (1 cmH(2)O, approximately 0.05 dyn/cm(2), 3-D), and after 6 h of exposure, expression of SM-MHC, smoothelin, and calponin were significantly reduced, while expression of alpha-SMA and SM22 were markedly enhanced. PD98059 (an ERK1/2 inhibitor) and heparinase III (an enzyme to cleave heparan sulfate) significantly blocked the effects of laminar Flow on gene expression, and also reversed the effects of Interstitial Flow on SM-MHC, smoothelin, and calponin, but enhanced Interstitial Flow-induced expression of alpha-SMA and SM22. SMCs and MFBs have similar responses to fluid Flow. Silencing ERK1/2 completely blocked the effects of both laminar Flow and Interstitial Flow on SMC marker gene expression. Western blotting showed that both types of Flows induced ERK1/2 activation that was inhibited by disruption of heparan sulfate proteoglycans (HSPGs). The results suggest that HSPG-mediated ERK1/2 activation is an important mechanotransduction pathway modulating SMC marker gene expression when SMCs and MFBs are exposed to Flow. Fluid Flow may be involved in vascular remodeling and lesion formation by affecting phenotypes of vascular wall cells. This study has implications in understanding the Flow-related mechanobiology in vascular lesion formation, tumor cell invasion, and stem cell differentiation.
Roger D Kamm - One of the best experts on this subject based on the ideXlab platform.
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Balance of Interstitial Flow magnitude and vascular endothelial growth factor concentration modulates three-dimensional microvascular network formation.
APL bioengineering, 2019Co-Authors: Yoshinori Abe, Roger D Kamm, Masafumi Watanabe, Seok Chung, Kazuo Tanishita, Ryo SudoAbstract:Hemodynamic and biochemical factors play important roles in critical steps of angiogenesis. In particular, Interstitial Flow has attracted attention as an important hemodynamic factor controlling the angiogenic process. Here, we applied a wide range of Interstitial Flow magnitudes to an in vitro three-dimensional (3D) angiogenesis model in a microfluidic device. This study aimed to investigate the effect of Interstitial Flow magnitude in combination with the vascular endothelial growth factor (VEGF) concentration on 3D microvascular network formation. Human umbilical vein endothelial cells (HUVECs) were cultured in a series of Interstitial Flow generated by 2, 8, and 25 mmH2O. Our findings indicated that Interstitial Flow significantly enhanced vascular sprout formation, network extension, and the development of branching networks in a magnitude-dependent manner. Furthermore, we demonstrated that the proangiogenic effect of Interstitial Flow application could not be substituted by the increased VEGF concentration. In addition, we found that HUVECs near vascular sprouts significantly elongated in >8 mmH2O conditions, while activation of Src was detected even in 2 mmH2O conditions. Our results suggest that the balance between the Interstitial Flow magnitude and the VEGF concentration plays an important role in the regulation of 3D microvascular network formation in vitro.
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Interstitial Flow promotes macrophage polarization toward an M2 phenotype.
Molecular biology of the cell, 2018Co-Authors: Jean Carlos Serrano, Hao Xing, Tara A. Lee, Hesham Azizgolshani, Muhammad H. Zaman, Roger D KammAbstract:Tumor tissues are characterized by an elevated Interstitial fluid Flow from the tumor to the surrounding stroma. Macrophages in the tumor microenvironment are key contributors to tumor progression. While it is well established that chemical stimuli within the tumor tissues can alter macrophage behaviors, the effects of mechanical stimuli, especially the Flow of Interstitial fluid in the tumor microenvironment, on macrophage phenotypes have not been explored. Here, we used three-dimensional biomimetic models to reveal that macrophages can sense and respond to pathophysiological levels of Interstitial fluid Flow reported in tumors (∼3 µm/s). Specifically, Interstitial Flow (IF) polarizes macrophages toward an M2-like phenotype via integrin/Src-mediated mechanotransduction pathways involving STAT3/6. Consistent with this Flow-induced M2 polarization, macrophages treated with IF migrate faster and have an enhanced ability to promote cancer cell migration. Moreover, IF directs macrophages to migrate against the Flow. Since IF emanates from the tumor to the surrounding stromal tissues, our results suggest that IF could not only induce M2 polarization of macrophages but also recruit these M2 macrophages toward the tumor masses, contributing to cancer cell invasion and tumor progression. Collectively, our study reveals that IF could be a critical regulator of tumor immune environment.
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Abstract A49: Tumor-associated Interstitial Flow promotes macrophage migration and pro-metastatic M2 phenotype in 3D ECM
Tumor Mechanobiology, 2017Co-Authors: Hao Xing, Tara A. Lee, Hesham Azizgolshani, Roger D KammAbstract:The growth of solid tumor is often accompanied by an increase in the Flow of Interstitial fluid (Interstitial Flow) from the center of the tumor to the surrounding stroma. Recent studies have shown that elevated Interstitial Flow can influence the migration of cancer cells and fibroblasts. Despite these findings, the effects of Interstitial Flow on macrophages, one of the key metastasis-promoting tumor-associated stromal cell types, have not been explored. We used microfluidic and transwell Flow assays to investigate how high level of Interstitial Flow (~3 μm/s) often observed in tumor affect macrophage migration and protein expression. We found that Interstitial Flow not only enhanced macrophage migration speed, but also directed macrophages to migrate against the Flow in 3D extracellular matrix. Moreover, we discovered that Interstitial Flow activated Akt and FAK, two kinases that are crucial for cell migration. Interestingly, Interstitial Flow also up-regulated macrophage expression of M2 markers Arg I and TGFβ through the activation of JAK1-STAT3/6 pathway. Finally, macrophages treated with Interstitial Flow were shown to have enhanced abilities to promote cancer cell migration and protrusion formation. Taken together, these results suggest that Interstitial Flow can promote metastasis through its effects on macrophages. Specifically, since macrophages migrate against the Flow, Interstitial Flow, which emanates from the tumor, can act as a mechanical stimulus to recruit macrophages into tumor tissues. Moreover, Interstitial Flow can activate macrophages and polarize them toward pro-metastatic M2 phenotype to further promote tumor metastasis and immunosuppression. Citation Format: Ran Li, Hao Xing, Tara A. Lee, Hesham Azizgolshani, Roger D. Kamm. Tumor-associated Interstitial Flow promotes macrophage migration and pro-metastatic M2 phenotype in 3D ECM. [abstract]. In: Proceedings of the AACR Special Conference on Engineering and Physical Sciences in Oncology; 2016 Jun 25-28; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2017;77(2 Suppl):Abstract nr A49.
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A three-dimensional microfluidic tumor cell migration assay to screen the effect of anti-migratory drugs and Interstitial Flow
Microfluidics and Nanofluidics, 2012Co-Authors: Johann Kalchman, Roger D Kamm, Seok Chung, Kazuo Tanishita, Shingo Fujioka, Yamato Kikkawa, Toshihiro Mitaka, Ryo SudoAbstract:Most anti-cancer drug screening assays are currently performed in two dimensions, on flat, rigid surfaces. However, there are increasing indications that three-dimensional (3D) platforms provide a more realistic setting to investigate accurate morphology, growth, and sensitivity of tumor cells to chemical factors. Moreover, Interstitial Flow plays a pivotal role in tumor growth. Here, we present a microfluidic 3D platform to investigate behaviors of tumor cells in Flow conditions with anti-migratory compounds. Our results show that Interstitial Flow and its direction have significant impact on migration and growth of hepatocellular carcinoma cell lines such as HepG2 and HLE. In particular, HepG2/HLE cells tend to migrate against Interstitial Flow, and their growth increases in Interstitial Flow conditions regardless of the Flow direction. Furthermore, this migratory activity of HepG2 cells is enhanced when they are co-cultured with human umbilical vein endothelial cells. We also found that migration activity of HepG2 cells attenuates under hypoxic conditions. In addition, the effect of Artemisinin, an anti-migratory compound, on HepG2 cells was quantitatively analyzed. The microfluidic 3D platform described here is useful to investigate more accurately the effect of anti-migratory drugs on tumor cells and the critical influence of Interstitial Flow than 2D culture models.
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Interstitial Flow influences direction of tumor cell migration through competing mechanisms
Proceedings of the National Academy of Sciences, 2011Co-Authors: William J Polacheck, Joseph L. Charest, Roger D KammAbstract:Interstitial Flow is the convective transport of fluid through tissue extracellular matrix. This creeping fluid Flow has been shown to affect the morphology and migration of cells such as fibroblasts, cancer cells, endothelial cells, and mesenchymal stem cells. A microfluidic cell culture system was designed to apply stable pressure gradients and fluid Flow and allow direct visualization of transient responses of cells seeded in a 3D collagen type I scaffold. We used this system to examine the effects of Interstitial Flow on cancer cell morphology and migration and to extend previous studies showing that Interstitial Flow increases the metastatic potential of MDA-MB-435S melanoma cells [Shields J, et al. (2007) Cancer Cell 11:526-538]. Using a breast carcinoma line (MDA-MB-231) we also observed cell migration along streamlines in the presence of Flow; however, we further demonstrated that the strength of the Flow as well as the cell density determined directional bias of migration along the streamline. In particular, we found that cells either at high seeding density or with the CCR-7 receptor inhibited migration against, rather than with the Flow. We provide further evidence that CCR7-dependent autologous chemotaxis is the mechanism that leads to migration with the Flow, but also demonstrate a competing CCR7-independent mechanism that causes migration against the Flow. Data from experiments investigating the effects of cell concentration, Interstitial Flow rate, receptor activity, and focal adhesion kinase phosphorylation support our hypothesis that the competing stimulus is integrin mediated. This mechanism may play an important role in development of metastatic disease.
Steven C George - One of the best experts on this subject based on the ideXlab platform.
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Low levels of physiological Interstitial Flow eliminate morphogen gradients and guide angiogenesis.
Angiogenesis, 2017Co-Authors: Venktesh S Shirure, Andrew Lezia, Arnold Tao, Luis F. Alonzo, Steven C GeorgeAbstract:Convective transport can significantly distort spatial concentration gradients. Interstitial Flow is ubiquitous throughout living tissue, but our understanding of how Interstitial Flow affects concentration gradients in biological processes is limited. Interstitial Flow is of particular interest for angiogenesis because pathological and physiological angiogenesis is associated with altered Interstitial Flow, and both Interstitial Flow and morphogen gradients (e.g., vascular endothelial growth factor, VEGF) can potentially stimulate and guide new blood vessel growth. We designed an in vitro microfluidic platform to simulate 3D angiogenesis in a tissue microenvironment that precisely controls Interstitial Flow and spatial morphogen gradients. The microvascular tissue was developed from endothelial colony forming cell-derived endothelial cells extracted from cord blood and stromal fibroblasts in a fibrin extracellular matrix. Pressure in the microfluidic lines was manipulated to control the Interstitial Flow. A mathematical model of mass and momentum transport, and experimental studies with fluorescently labeled dextran were performed to validate the platform. Our data demonstrate that at physiological Interstitial Flow (0.1–10 μm/s), morphogen gradients were eliminated within hours, and angiogenesis demonstrated a striking bias in the opposite direction of Interstitial Flow. The Interstitial Flow-directed angiogenesis was dependent on the presence of VEGF, and the effect was mediated by αvβ3 integrin. We conclude that under physiological conditions, growth factors such as VEGF and fluid forces work together to initiate and spatially guide angiogenesis.
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microfluidic device to control Interstitial Flow mediated homotypic and heterotypic cellular communication
Lab on a Chip, 2015Co-Authors: Luis F. Alonzo, Venktesh S Shirure, Monica L Moya, Steven C GeorgeAbstract:Tissue engineering can potentially recreate in vivo cellular microenvironments in vitro for an array of applications such as biological inquiry and drug discovery. However, the majority of current in vitro systems still neglect many biological, chemical, and mechanical cues that are known to impact cellular functions such as proliferation, migration, and differentiation. To address this gap, we have developed a novel microfluidic device that precisely controls the spatial and temporal interactions between adjacent three-dimensional cellular environments. The device consists of four interconnected microtissue compartments (~0.1 mm3) arranged in a square. The top and bottom pairs of compartments can be sequentially loaded with discrete cellularized hydrogels creating the opportunity to investigate homotypic (left to right or x-direction) and heterotypic (top to bottom or y-direction) cell–cell communication. A controlled hydrostatic pressure difference across the tissue compartments in both x and y direction induces Interstitial Flow and modulates communication via soluble factors. To validate the biological significance of this novel platform, we examined the role of stromal cells in the process of vasculogenesis. Our device confirms previous observations that soluble mediators derived from normal human lung fibroblasts (NHLFs) are necessary to form a vascular network derived from endothelial colony forming cell-derived endothelial cells (ECFC-ECs). We conclude that this platform could be used to study important physiological and pathological processes that rely on homotypic and heterotypic cell–cell communication.
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abstract 4806 angiogenesis is independently influenced by Interstitial Flow and concentration gradients of tumor secreted morphogens
Cancer Research, 2014Co-Authors: Venktesh S Shirure, Steven C GeorgeAbstract:Angiogenesis facilitates growth and entry of tumors into the systemic circulation leading to metastasis. Morphogens, such as vascular endothelial growth factor (VEGF), secreted by the tumors are believed to guide nearby vasculature towards the tumor. The distribution of morphogens in tissues is affected by Interstitial fluid Flow, which also exerts forces in the tissue microenvironment. Numerous cells in the tumor microenvironment can sense and respond to force, yet our understanding of the independent effect of Interstitial Flow in tumor angiogenesis is limited. This is due, in part, to inadequate experimental models that cannot decouple spatial morphogen concentrations and Interstitial Flow. Notably, control of these parameters is not possible in animal models, and 2D cell systems lack the complexity of in vivo tissue. We addressed these limitations by designing a microfluidic platform of polydimethlsiloxane (PDMS) polymer consisting of three parallel microporous tissue compartments ( Citation Format: Venktesh S. Shirure, Steven C. George. Angiogenesis is independently influenced by Interstitial Flow and concentration gradients of tumor secreted morphogens. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4806. doi:10.1158/1538-7445.AM2014-4806
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Abstract 3930: Recapitulating the microenvironment in vitro for comparative study of factors affecting tumor growth and vascularization
Tumor Biology, 2014Co-Authors: Luis F. Alonzo, Monica L Moya, Claire Robertson, Marian L. Waterman, Christopher C.w. Hughes, Steven C GeorgeAbstract:Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA Introduction: Tumor growth is dramatically affected by the microenvironment, including supporting cells such as the stroma and vasculature, mechanical factors, such as Interstitial Flow and extracellular matrix and aspects of the tumor mass itself, including shape. However, current tumor growth models, including xenograft models, 2D and/or simplistic 3D cultures, are unable to address these interactions in a high throughput human-derived system. We have developed a novel microfluidic platform that combines human derived perfused microvessels, stroma, and Interstitial Flow with 3D culture. This platform was used to quantitatively compare the role of these microenvironmental factors on tumor growth. Methods: A microfluidic device was fabricated consisting of two supply channels on either side of a central tissue compartment. The inner stromal compartment consists of normal human fibroblasts (NHLFs) and GFP-labeled human colorectal adenocarcinoma tumor cells, SW620, seeded in a fibrin matrix. To simulate a vasculogenic-like process, human cord blood endothelial colony forming cells endothelial cells (ECFC-ECs) were distributed throughout the stromal channel with the fibroblasts and tumor cells. Tumor growth rate and area was compared across day, Interstitial Flow rates and tumor shape (fractal dimension, perimeter to area) with ANOVA. Results: Cell viability within the device was maintained under Interstitial Flow conditions for a period of 21 days. Within one week of culture, microvessel formation and significant tumor growth into spheroids (n=636) were observed. On average, tumor growth rate was 26% ±62% per day with the highest growth rates observed on the first days. By day 7, many tumor masses had died off, with 2-3 large, fast growing tumors remaining per chamber. Highest tumor growth rates and areas were observed in tumor masses with a characteristic morphology of high perimeter to area and lower cohesion. Interstitial Flow rates ranging from essentially static to supraphysiologic were generated. Differences in tumor growth rates were not statistically significant across chambers with different mean Flow rates. To demonstrate intraluminal Flow within the vascular network, fluorescently labeled dextran (40, 70, and 150 kDa) was introduced into the microfluidic lines. Dextran was retained in the vessel network, and showed tumor cells residing in the intraluminal space of the formed vasculature. By day 14, the network eroded, as the tumor masses overgrew and encompassed more than 60% of the chamber volume. Discussion: We have developed a novel 3D microfluidic system of the tumor microenvironment that features perfused capillaries and controlled Interstitial Flow. Tumor growth was affected by tumor characteristic shape in this model though Interstitial Flow appeared to play a lesser role. Vascular development was observed and its interaction with tumor growth will be analyzed in future work. Citation Format: Luis F. Alonzo, Claire J. Robertson, Monica L. Moya, Marian L. Waterman, Christopher C. Hughes, Steven C. George. Recapitulating the microenvironment in vitro for comparative study of factors affecting tumor growth and vascularization. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3930. doi:10.1158/1538-7445.AM2014-3930
