scholarly journals Matrix mechanics regulates epithelial defence against cancer by tuning dynamic localization of filamin

2022 ◽  
Vol 13 (1) ◽  
Author(s):  
Shilpa P. Pothapragada ◽  
Praver Gupta ◽  
Soumi Mukherjee ◽  
Tamal Das
Keyword(s):  
Normal Cell ◽  
Transformed Cells ◽  
Matrix Stiffness ◽  
Loss Of Function ◽  
Single Population ◽  
Soft Matrix ◽  
Matrix Mechanics ◽  
Initial Stage ◽  
Protein Tracking ◽  
Cell Interface

AbstractIn epithelia, normal cells recognize and extrude out newly emerged transformed cells by competition. This process is the most fundamental epithelial defence against cancer, whose occasional failure promotes oncogenesis. However, little is known about what factors determine the success or failure of this defence. Here we report that mechanical stiffening of extracellular matrix attenuates the epithelial defence against HRasV12-transformed cells. Using photoconversion labelling, protein tracking, and loss-of-function mutations, we attribute this attenuation to stiffening-induced perinuclear sequestration of a cytoskeletal protein, filamin. On soft matrix mimicking healthy epithelium, filamin exists as a dynamically single population, which moves to the normal cell-transformed cell interface to initiate the extrusion of transformed cells. However, on stiff matrix mimicking fibrotic epithelium, filamin redistributes into two dynamically distinct populations, including a new perinuclear pool that cannot move to the cell-cell interface. A matrix stiffness-dependent differential between filamin-Cdc42 and filamin-perinuclear cytoskeleton interaction controls this distinctive filamin localization and hence, determines the success or failure of epithelial defence on soft versus stiff matrix. Together, our study reveals how pathological matrix stiffening leads to a failed epithelial defence at the initial stage of oncogenesis.

scholarly journals Matrix mechanics regulates epithelial defence against cancer by tuning dynamic localization of filamin

2020 ◽  
Author(s):  
Shilpa Pothapragada ◽  
Praver Gupta ◽  
Soumi Mukherjee ◽  
Tamal Das
Keyword(s):  
Transformed Cells ◽  
Loss Of Function ◽  
Single Population ◽  
Soft Matrix ◽  
Transformed Cell ◽  
Matrix Mechanics ◽  
Initial Stage ◽  
Protein Tracking ◽  
Cell Interface ◽  
Cell Extrusion

Abstract In epithelia, normal cells recognize and extrude out newly emerged transformed cells by competition. This process is the most fundamental epithelial defence against cancer, whose occasional failure promotes oncogenesis. However, little is known about what factors determine the success or failure of this defence. Here we report that mechanical stiffening of extracellular matrix attenuates the epithelial defence against activated HRasV12-transformed cells. Using photoconversion labelling, protein tracking, and loss-of-function mutations, we attribute this attenuation to stiffening-induced perinuclear sequestration of a cytoskeletal protein, filamin. On soft matrix mimicking healthy epithelium, filamin exists as a dynamically single population, which moves to the normal cell-transformed cell interface to initiate transformed cell-extrusion. But, on stiff matrix mimicking fibrotic epithelium, filamin redistributes into two dynamically distinct populations, including a new perinuclear pool, which cannot move to the cell-cell interface. A tug-of-war between filamin-Cdc42 and filamin-perinuclear cytoskeleton interactions controls this differential filamin localization and hence, determines the success or failure of epithelial defence on soft versus stiff matrix. Together, our study reveals how pathological matrix stiffening leads to a failed epithelial defence at the initial stage of oncogenesis.

scholarly journals Serum-free growth of normal and transformed fibroblasts in milk: differential requirements for fibronectin.

1981 ◽  
Vol 88 (2) ◽  
pp. 294-300 ◽  
Author(s):  
K S Steimer ◽  
M Klagsbrun
Keyword(s):  
Cell Lines ◽  
Normal Cell ◽  
Transformed Cells ◽  
Temperature Sensitive ◽  
Fibroblastic Cell ◽  
Serum Free ◽  
Transformed Fibroblasts ◽  
Cell Strains ◽  
Free Growth ◽  
Fibroblastic Cells

Bovine milk may be used as a supplement for the serum-free growth of certain fibroblastic cells in culture. The growth properties of three representative cell types in milk-supplemented medium were examined; fibroblastic cell strains, fibroblastic cell lines, and transformed fibroblasts. Transformed fibroblasts, which included RNA and DNA tumor virus-transformed cells and carcinogen-transformed cells, grew in milk. Instead of growing attached to the culture dishes, as they normally do in serum, transformed fibroblasts grew in milk as large clusters in suspension. In contrast, nontransformed fibroblastic cell strains and cell lines did not grow in milk-supplemented medium. Fibroblasts transformed by a temperature-sensitive transformation mutant of Rous sarcoma virus were temperature-sensitive for growth in milk. The failure of cells to adhere to the substratum in milk-supplemented medium suggested that milk might be deficient in attachment factors for fibroblasts. When the attachment of fibroblastic cells in milk-supplemented medium was facilitated by pretreating culture dishes with fibronectin, (a) transformed cells grew attached rather than in suspension, (b) normal cell lines attached and grew to confluence, and (c) normal cell strains adhered and survived but did not exhibit appreciable cell proliferation.

scholarly journals Matrix stiffness and blood pressure together regulate vascular smooth muscle cell phenotype switching and cofilin dependent podosome formation

2020 ◽  
Author(s):  
Brian Sit ◽  
Zhen Feng ◽  
Ioannis Xanthis ◽  
Emilie Marhuenda ◽  
Simona Zingaro ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Phenotypic Switching ◽  
Cell Phenotype ◽  
Matrix Stiffness ◽  
Cellular Mechanics ◽  
Soft Matrix ◽  
Phenotype Switch ◽  
The Impact ◽  
Progression Of Atherosclerosis

AbstractVascular smooth muscle cells (VSMCs) play a central role in the onset and progression of atherosclerosis. In pre-atherosclerotic lesions, VSMCs switch from a contractile to a synthetic phenotype and subsequently remodel the microenvironment, leading to further disease progression. Ageing and associated mechanical changes of the extracellular matrix as well as hypertension are major risk of atherosclerosis. Consequently, we sought here to systematically study the impact of mechanical and chemical stimulations on VSMC phenotypic switching. We find that the hemodynamic pressure and matrix stiffness have overlapping effects and together contribute to the phenotypic changes in cellular mechanics, podosome formation, and matrix degradation. We further identify cofilin as a key modulator of the mechanosensitive phenotype switch, which is regulated through Ca2+/slingshot-dependent pressure sensing and RhoA/ROCK-dependent stiffness sensing pathways. Altogether, microenvironment stimulations of high pressure and soft matrix collectively promote the cofilin activity, VSMC migration, and the early progression of atherosclerosis.

scholarly journals Src-transformed cells hijack mitosis to extrude from the epithelium

2018 ◽  
Author(s):  
Katarzyna A. Anton ◽  
Mihoko Kajita ◽  
Rika Narumi ◽  
Yasuyuki Fujita ◽  
Masazumi Tada
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Progression ◽  
Apical Part ◽  
Transformed Cells ◽  
Cycle Progression ◽  
Cellular Processes ◽  
Initial Stage ◽  
Apical Extrusion

AbstractAt the initial stage of carcinogenesis single mutated cells appear within an epithelium. Mammalian in vitro experiments show that potentially cancerous cells undergo live apical extrusion from normal monolayers. However, the mechanism underlying this process in vivo remains poorly understood. Mosaic expression of the oncogene vSrc in a simple epithelium of the early zebrafish embryo results in apical extrusion of transformed cells. Here we find that during extrusion components of the cytokinetic ring are recruited to adherens junctions of transformed cells, stimulating formation of a misoriented pseudo cytokinetic ring. During extrusion, the ring constricts and separates the basal from the apical part of the cell releasing both from the epithelium. This process requires cell cycle progression and occurs immediately after vSrc-transformed cell enters mitosis. To achieve extrusion, vSrc coordinates cell cycle progression, junctional integrity, cell survival and apicobasal polarity. Without vSrc, modulating these cellular processes reconstitutes vSrc-like extrusion, confirming their sufficiency for this process.

scholarly journals INFLUENCE OF LEAFLET’S MATRIX STIFFNESS AND FIBER ORIENTATION ON THE OPENING DYNAMICS OF A PROSTHETIC TRILEAFLET HEART VALVE

2017 ◽  
Vol 17 (06) ◽  
pp. 1750096 ◽  
Author(s):  
ANDREA AVANZINI
Keyword(s):  
Fiber Orientation ◽  
Mechanical Response ◽  
Strain Energy Function ◽  
Effective Orifice Area ◽  
Matrix Stiffness ◽  
Soft Matrix ◽  
Long Time ◽  
Valve Dynamics ◽  
Lower Maximum ◽  
Opening Phase

Biological valves are employed for aortic valve substitution since a long time but there is a growing effort toward the development of new engineered tissues, in which the complex mechanical response of native leaflets is replicated using composite materials consisting of a soft matrix with embedded reinforcing fibers. The main goal of the present study is to investigate the influence that variations on fiber orientation and matrix stiffness may have on valve dynamics. To this aim, a fluid–structure interaction (FSI) model of a trileaflet valve was implemented in which the opening phase was simulated and leaflet matrix stiffness and fiber orientation were varied in the framework of an anisotropic hyperelastic strain energy function. Results show that both parameters may affect significantly transvalvular pressure gradient and effective orifice area (EOA). For the opening phase of the valve examined, less favorable flow conditions were found when preferred fiber orientation is circumferential, due to lower maximum EOA achievable. Such configuration in combination with stiffer matrix may result in significant degradation of valve performances. Overall fiber orientation can potentially be taylored to optimize valve dynamics, provided also structural aspects that may be prominent in the closure phase, are considered.

scholarly journals TRPV4 Plays a Role in Matrix Stiffness-Induced Macrophage Polarization

2020 ◽  
Vol 11 ◽  
Author(s):  
Bidisha Dutta ◽  
Rishov Goswami ◽  
Shaik O. Rahaman
Keyword(s):  
Transient Receptor Potential ◽  
Macrophage Polarization ◽  
Transient Receptor Potential Vanilloid ◽  
Polarization Spectrum ◽  
Dependent Manner ◽  
Matrix Stiffness ◽  
Soft Matrix ◽  
Genetic Ablation ◽  
M1 Macrophage

Phenotypic polarization of macrophages is deemed essential in innate immunity and various pathophysiological conditions. We have now determined key aspects of the molecular mechanism by which mechanical cues regulate macrophage polarization. We show that Transient Receptor Potential Vanilloid 4 (TRPV4), a mechanosensitive ion channel, mediates substrate stiffness-induced macrophage polarization. Using atomic force microscopy, we showed that genetic ablation of TRPV4 function abrogated fibrosis-induced matrix stiffness generation in skin tissues. We have determined that stiffer skin tissue promotes the M1 macrophage subtype in a TRPV4-dependent manner; soft tissue does not. These findings were further validated by our in vitro results which showed that stiff matrix (50 kPa) alone increased expression of macrophage M1 markers in a TRPV4-dependent manner, and this response was further augmented by the addition of soluble factors; neither of which occurred with soft matrix (1 kPa). A direct requirement for TRPV4 in M1 macrophage polarization spectrum in response to increased stiffness was evident from results of gain-of-function assays, where reintroduction of TRPV4 significantly upregulated the expression of M1 markers in TRPV4 KO macrophages. Together, these data provide new insights regarding the role of TRPV4 in matrix stiffness-induced macrophage polarization spectrum that may be explored in tissue engineering and regenerative medicine and targeted therapeutics.

Regulation of proximal tubular cell differentiation and proliferation in primary culture by matrix stiffness and ECM components

2014 ◽  
Vol 307 (6) ◽  
pp. F695-F707 ◽  
Author(s):  
Wan-Chun Chen ◽  
Hsi-Hui Lin ◽  
Ming-Jer Tang
Keyword(s):  
Cell Differentiation ◽  
Transforming Growth Factor ◽  
Primary Cultures ◽  
Type I ◽  
Matrix Stiffness ◽  
Mesenchymal Transition ◽  
Soft Matrix ◽  
The Matrix ◽  
Proximal Tubular ◽  
Differentiation And Proliferation

To explore whether matrix stiffness affects cell differentiation, proliferation, and transforming growth factor (TGF)-β1-induced epithelial-mesenchymal transition (EMT) in primary cultures of mouse proximal tubular epithelial cells (mPTECs), we used a soft matrix made from monomeric collagen type I-coated polyacrylamide gel or matrigel (MG). Both kinds of soft matrix benefited primary mPTECs to retain tubular-like morphology with differentiation and growth arrest and to evade TGF-β1-induced EMT. However, the potent effect of MG on mPTEC differentiation was suppressed by glutaraldehyde-induced cross-linking and subsequently stiffening MG or by an increasing ratio of collagen in the soft mixed gel. Culture media supplemented with MG also helped mPTECs to retain tubular-like morphology and a differentiated phenotype on stiff culture dishes as soft MG did. We further found that the protein level and activity of ERK were scaled with the matrix stiffness. U-0126, a MEK inhibitor, abolished the stiff matrix-induced dedifferentiation and proliferation. These data suggest that the ERK signaling pathway plays a vital role in matrix stiffness-regulated cell growth and differentiation. Taken together, both compliant property and specific MG signals from the matrix are required for the regulation of epithelial differentiation and proliferation. This study provides a basic understanding of how physical and chemical cues derived from the extracellular matrix regulate the physiological function of proximal tubules and the pathological development of renal fibrosis.

scholarly journals Control of matrix stiffness promotes endodermal lineage specification by regulating SMAD2/3 via lncRNA LINC00458

2020 ◽  
Vol 6 (6) ◽  
pp. eaay0264 ◽  
Author(s):  
Yu-Fan Chen ◽  
Yi-Shuan J. Li ◽  
Chih-Hung Chou ◽  
Men Yee Chiew ◽  
Hsien-Da Huang ◽  
...  
Keyword(s):  
Young’S Modulus ◽  
Young's Modulus ◽  
Noncoding Rnas ◽  
Specific Gene ◽  
Sequencing Analysis ◽  
Matrix Stiffness ◽  
Loss Of Function ◽  
Lineage Specification ◽  
Soft Substrate ◽  
Extrinsic Factors

During endoderm formation, cell identity and tissue morphogenesis are tightly controlled by cell-intrinsic and cell-extrinsic factors such as biochemical and physical inputs. While the effects of biochemical factors are well studied, the physical cues that regulate cell division and differentiation are poorly understood. RNA sequencing analysis demonstrated increases of endoderm-specific gene expression in hPSCs cultured on soft substrate (Young’s modulus, 3 ± 0.45 kPa) in comparison with hard substrate (Young’s modulus, 165 ± 6.39 kPa). Further analyses revealed that multiple long noncoding RNAs (lncRNAs) were up-regulated on soft substrate; among them, LINC00458 was identified as a stiffness-dependent lncRNA specifically required for hPSC differentiation toward an early endodermal lineage. Gain- and loss-of-function experiments confirmed that LINC00458 is functionally required for hPSC endodermal lineage specification induced by soft substrates. Our study provides evidence that mechanical cues regulate the expression of LINC00458 and induce differentiation of hPSC into hepatic lineage progenitors.

Stimulated Virus Production in Vinblastine and Cytochalasin-Induced Multinucleate Cells

1970 ◽  
Vol 28 ◽  
pp. 186-187
Author(s):  
Krishan Awtar
Keyword(s):  
Cell Division ◽  
Normal Cell ◽  
Virus Production ◽  
Multinucleate Cells ◽  
Daughter Cells ◽  
Cell Divisions ◽  
Nuclear Membranes ◽  
Division 2 ◽  
Vinblastine Sulfate

Exposure of cells to low sublethal but mitosis-arresting doses of vinblastine sulfate (Velban) results in the initial arrest of cells in mitosis followed by their subsequent return to an “interphase“-like stage. A large number of these cells reform their nuclear membranes and form large multimicronucleated cells, some containing as many as 25 or more micronuclei (1). Formation of large multinucleate cells is also caused by cytochalasin, by causing the fusion of daughter cells at the end of an otherwise .normal cell division (2). By the repetition of this process through subsequent cell divisions, large cells with 6 or more nuclei are formed.

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