Spatial Proliferation of Epithelial Cells Is Regulated by E-Cadherin Force

Mohan, Ayush; Schlue, Kyle T.; Kniffin, Alex F.; Mayer, Carl R.; Duke, Ashley; Narayanan, Vani; Arsenovic, Paul T.; Bathula, Kranthidhar; Danielsson, Brooke E; Dumbali, Sandeep; Maruthamuthu, Venkat; Conway, Daniel E.

doi:10.1016/j.bpj.2018.07.030

Cited by 24 publications

(28 citation statements)

References 34 publications

(46 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To remedy these shortcomings, we developed two cell stretching apparatuses which can apply uniaxial strains of up to 300% in one dimension or biaxial strains of up to 60% in both dimensions. We previously used these devices to apply mechanical forces to adherens junctions 16 and desmosomes, 6 two principal structures of cell-cell contacts. Both devices are suitable for in situ imaging, and the components of which can be made using a 3D printer and assembled using hand tools.…”

Section: Introductionmentioning

confidence: 99%

Characterization of 3D Printed Stretching Devices for Imaging Force Transmission in Live-Cells

et al. 2019

Self Cite

View full text Add to dashboard Cite

Introduction-Cell stretch is a method which can rapidly apply mechanical force through cell-matrix and cell-cell adhesions and can be utilized to better understand underlying biophysical questions related to intracellular force transmission and mechanotransduction. Methods-3D printable stretching devices suitable for livecell fluorescent imaging were designed using finite element modeling and validated experimentally. These devices were then used along with FRET based nesprin-2G force sensitive biosensors as well as live cell fluorescent staining to understand how the nucleus responds to externally applied mechanical force in cells with both intact LINC (linker of nucleoskeleton and cytoskeleton) complex and cells with the LINC complex disrupted using expression of dominant negative KASH protein. Results-The devices were shown to provide a larger strain ranges (300% uniaxial and 60% biaxial) than currently available commercial or academic designs we are aware of. Under uniaxial deformation, the deformation of the nucleus of NIH 3T3 cells per unit of imposed cell strain was shown to be approximately 50% higher in control cells compared to cells with a disrupted LINC complex. Under biaxial deformation, MDCK II cells showed permanent changes in the nuclear morphology as well as actin organization upon unloading, indicating that failure, plastic deformation, or remodeling of the cytoskeleton is occurring in response to the applied stretch. Conclusion-Development and open distribution of low-cost, 3D-printable uniaxial and biaxial cell stretching devices compatible with live-cell fluorescent imaging allows a wider range of researchers to investigate mechanical influences on biological questions with only a minimal investment of resources.

show abstract

Section: Introductionmentioning

confidence: 99%

Characterization of 3D Printed Stretching Devices for Imaging Force Transmission in Live-Cells

et al. 2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…Though adult cardiomyocyte has lost the ability to entry cell cycle, cardiomyocyte from embryonic and neonatal mammal is capable of proliferating . Prior experiments have shown that externally applied forces result in increased proliferation in an E‐cadherin force‐dependent manner . In this study, we demonstrated that simulated microgravity and hypergravity can influence the cardiomyocyte proliferation in different extent.…”

Section: Discussionmentioning

confidence: 61%

Alteration of calcium signalling in cardiomyocyte induced by simulated microgravity and hypergravity

Liu

Zhong

Zhou³

et al. 2020

Cell Proliferation

View full text Add to dashboard Cite

Objectives Cardiac Ca2+ signalling plays an essential role in regulating excitation‐contraction coupling and cardiac remodelling. However, the response of cardiomyocytes to simulated microgravity and hypergravity and the effects on Ca2+ signalling remain unknown. Here, we elucidate the mechanisms underlying the proliferation and remodelling of HL‐1 cardiomyocytes subjected to rotation‐simulated microgravity and 4G hypergravity. Materials and Methods The cardiomyocyte cell line HL‐1 was used in this study. A clinostat and centrifuge were used to study the effects of microgravity and hypergravity, respectively, on cells. Calcium signalling was detected with laser scanning confocal microscopy. Protein and mRNA levels were detected by Western blotting and real‐time PCR, respectively. Wheat germ agglutinin (WGA) staining was used to analyse cell size. Results Our data showed that spontaneous calcium oscillations and cytosolic calcium concentration are both increased in HL‐1 cells after simulated microgravity and 4G hypergravity. Increased cytosolic calcium leads to activation of calmodulin‐dependent protein kinase II/histone deacetylase 4 (CaMKII/HDAC4) signalling and upregulation of the foetal genes ANP and BNP, indicating cardiac remodelling. WGA staining indicated that cell size was decreased following rotation‐simulated microgravity and increased following 4G hypergravity. Moreover, HL‐1 cell proliferation was increased significantly under hypergravity but not rotation‐simulated microgravity. Conclusions Our study demonstrates for the first time that Ca2+/CaMKII/HDAC4 signalling plays a pivotal role in myocardial remodelling under rotation‐simulated microgravity and hypergravity.

show abstract

“…We next sought to compare these with experimentally-measured junction forces. To measure cell-cell junction forces experimentally, Madin-Darby Canine Kidney Cells (MDCKII) cells were stably transfected with a full-length E-cadherin force sensor, as previously described [18]. Briefly, the force sensor consists of two fluorophores coupled by a polypeptide that exhibits elasticity.…”

Section: Resultsmentioning

confidence: 99%

A predictive model of intercellular tension and cell-matrix mechanical interactions in a multicellular geometry

Scott

Griggs

Narayanan

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

Epithelial cells form continuous sheets of cells that exist in tensional homeostasis.Homeostasis is maintained through cell-to-cell adhesions that distribute tension and balance forces between cells and their underlying matrix. Disruption of tensional homeostasis can lead to Epithelial-Mesenchymal Transition (EMT), which is a transdifferentiation process in which epithelial cells adopt a mesenchymal phenotype, where cell-cell adhesion is lost and individual cell migration is acquired. This process is critical during embryogenesis and wound healing, but is also dysregulated in many disease states. To further understand the role of intercellular tension in spatial patterning of epithelial cell monolayers, we developed a multicellular computational model of cell-cell and cell-substrate forces. This work builds on a hybrid Cellular Potts-finite element model to evaluate cell-matrix mechanical feedback of an adherent multicellular cluster. Thermodynamically-constrained cells migrate by generating traction forces on a finite element substrate to minimize the total energy of the system. Junctional forces at cell-cell contacts balance these traction forces, thereby producing a mechanically stable epithelial monolayer. Simulations were compared to in vitro experiments using fluorescence-based junction force sensors in clusters of cells undergoing EMT. Results indicate that the multicellular CPM model can reproduce many aspects of EMT, including epithelial monolayer formation dynamics, changes in cell geometry, and spatial patterning of cell geometry and cell-cell forces in an epithelial colony. Author summaryEpithelial cells line all organs of the human body and act as a protective barrier by forming a continuous sheet. These cells exert force on both their neighboring cells as well as the underlying extracellular matrix, which is a network of proteins that creates the structure of tissues. Here we develop a model that encompasses both cell-cell forces and cell-matrix forces in an epithelial cell sheet. The model accounts for cell migration and proliferation, and regulates how cell-cell adhesions are formed. We demonstrate how the interplay between cell-cell forces and cell-matrix forces can regulate the formation of July 9, 2019 1/27 the epithelial cell sheet, the organization of cells within the sheet, and the pattern of cell geometries and cell forces within the sheet. We compare computational results with experiments in which epithelial cell sheets are disrupted and cell-cell junction forces are measured, and demonstrate that the model captures many aspects of epithelial cell dynamics observed experimentally. 42 cellular geometry. This results in a pattern of traction forces directed towards the cell 43 centroid and proportional to their distance from the cell centroid. These traction forces 44 generate substrate strains which, in addition to cell-cell and cell-matrix interactions, 45 July 9, 2019 2/27 impose a thermodynamic constraint and govern the dynamics of individual cells in the 46 CPM. In the current work, we ...

show abstract

Spatial Proliferation of Epithelial Cells Is Regulated by E-Cadherin Force

Cited by 24 publications

References 34 publications

Characterization of 3D Printed Stretching Devices for Imaging Force Transmission in Live-Cells

Characterization of 3D Printed Stretching Devices for Imaging Force Transmission in Live-Cells

Alteration of calcium signalling in cardiomyocyte induced by simulated microgravity and hypergravity

A predictive model of intercellular tension and cell-matrix mechanical interactions in a multicellular geometry

Contact Info

Product

Resources

About