Traction force microscopy in rapidly moving cells reveals separate roles for ROCK and MLCK in the mechanics of retraction

Morin, Timothy R.; Ghassem-Zadeh, Sean A.; Lee, Juliet

doi:10.1016/j.yexcr.2014.04.015

Cited by 13 publications

(10 citation statements)

References 53 publications

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“…TFM revealed that the average cellular force was 125 Pa (range 21-349 Pa) (Fig. S2A,B), similar to previous reports of stromal cells (Munevar et al, 2001;Morin et al, 2014). The range of force generation by myofibroblasts suggests great heterogeneity in this population, with some cells being highly contractile and others exerting far weaker force.…”

Section: Results and Discussion Mapping The Mechanical Landscape In Hsupporting

confidence: 87%

Single cell force profiling of human myofibroblasts reveals a biophysical spectrum of cell states

et al. 2020

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Mechanical force is a fundamental regulator of cell phenotype. Myofibroblasts are central mediators of fibrosis, a major unmet clinical need characterised by the deposition of excessive matrix proteins. Traction forces of myofibroblasts play a key role in remodelling the matrix and modulate the activities of embedded stromal cells. Here, we employ a combination of unsupervised computational analysis, cytoskeletal profiling and single cell traction force microscopy as a functional readout to uncover how the complex spatiotemporal dynamics and mechanics of living human myofibroblast shape sub-cellular profiling of traction forces in fibrosis. We resolve distinct biophysical communities of myofibroblasts, and our results provide a new paradigm for studying functional heterogeneity in human stromal cells.

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Section: Results and Discussion Mapping The Mechanical Landscape In Hsupporting

confidence: 87%

Single cell force profiling of human myofibroblasts reveals a biophysical spectrum of cell states

et al. 2020

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“…To further directly assess cell contractility in Acvr1 +/+ and Acvr1 R206H/+ cells, we used traction force microscopy and determined that Acvr1 R206H/+ cells exert a higher traction stress on their substrates and generate a higher total force per cell (Fig. A ).…”

Section: Resultsmentioning

confidence: 99%

“…Traction force microscopy evaluates the extent to which a cell deforms its underlying ECM‐coated substrate. From this, one can calculate the total force generated by a single cell and the average traction stress (force per unit area) it applies to the substrate . A higher average traction stress indicates that a cell is in a more contractile state .…”

Section: Discussionmentioning

confidence: 99%

Elevated BMP and Mechanical Signaling Through YAP1/RhoA Poises FOP Mesenchymal Progenitors for Osteogenesis

Stanley

Heo

Mauck

et al. 2019

Journal of Bone and Mineral Research

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Fibrodysplasia ossificans progressiva (FOP) is a rare genetic disease characterized by the formation of extraskeletal bone, or heterotopic ossification (HO), in soft connective tissues such as skeletal muscle. All familial and sporadic cases with a classic clinical presentation of FOP carry a gain-of-function mutation (R206H; c.617 G > A) in ACVR1, a cell surface receptor that mediates bone morphogenetic protein (BMP) signaling. The BMP signaling pathway is recognized for its chondro/osteogenic-induction potential, and HO in FOP patients forms ectopic but qualitatively normal endochondral bone tissue through misdirected cell fate decisions by tissue-resident mesenchymal stem cells. In addition to biochemical ligand-receptor signaling, mechanical cues from the physical environment are transduced to activate intracellular signaling, a process known as mechanotransduction, and can influence cell fates. Utilizing an established mesenchymal stem cell model of mouse embryonic fibroblasts (MEFs) from the Acvr1 R206H/+ mouse model that mimics the human disease, we demonstrated that activation of the mechanotransductive effectors Rho/ROCK and YAP1 are increased in Acvr1 R206H/+ cells. We show that on softer substrates, a condition associated with low mechanical signaling, the morphology of Acvr1 R206H/+ cells is similar to the morphology of control Acvr1 +/+ cells on stiffer substrates, a condition that activates mechanotransduction. We further determined that Acvr1 R206H/+ cells are poised for osteogenic differentiation, expressing increased levels of chondro/osteogenic markers compared with Acvr1 +/+ cells. We also identified increased YAP1 nuclear localization in Acvr1 R206H/+ cells, which can be rescued by either BMP inhibition or Rho antagonism. Our results establish RhoA and YAP1 signaling as modulators of mechanotransduction in FOP and suggest that aberrant mechanical signals, combined with and as a result of the increased BMP pathway signaling through mutant ACVR1, lead to misinterpretation of the cellular microenvironment and a heightened sensitivity to mechanical stimuli that promotes commitment of Acvr1 R206H/+ progenitor cells to chondro/osteogenic lineages. Generation of mouse embryonic fibroblasts and cell cultureKnock-in Acvr1 R206H/+ MEFs were isolated at embryonic day 13.5 (E13.5) as previously described (43) from mice described in Chakkalakal and colleagues. (41) Genotypes of cell lines (MEFs) and presence of the R206H mutation were confirmed by PCR (primers in Supplemental Table S1). Acvr1 +/+ controls were littermates that did not contain an Acvr1 R206H allele. Cells were cultured in Dulbecco's modified Eagle's medium, high glucose (DMEM, Gibco, Thermo Fisher Scientific,

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“…While Traction Force Microscopy itself identifies absolute cell stresses and stress heat maps for single cells or cell clusters, TFM in combination with cell imaging has been used to track and correlate changes in cytoskeletal and associated signals with cell mobility [ 83 , 84 ], directionality of migration [ 58 , 60 , 82 , 85 ], heterogeneity of cell traction associated with proliferation [ 82 , 86 ] and stem cell fate [ 31 , 87 , 88 ].…”

Section: Tools To Study Cellular Mechanotransductionmentioning

confidence: 99%

Biophysical Tools to Study Cellular Mechanotransduction

2017

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The cell membrane is the interface that volumetrically isolates cellular components from the cell’s environment. Proteins embedded within and on the membrane have varied biological functions: reception of external biochemical signals, as membrane channels, amplification and regulation of chemical signals through secondary messenger molecules, controlled exocytosis, endocytosis, phagocytosis, organized recruitment and sequestration of cytosolic complex proteins, cell division processes, organization of the cytoskeleton and more. The membrane’s bioelectrical role is enabled by the physiologically controlled release and accumulation of electrochemical potential modulating molecules across the membrane through specialized ion channels (e.g., Na+, Ca2+, K+ channels). The membrane’s biomechanical functions include sensing external forces and/or the rigidity of the external environment through force transmission, specific conformational changes and/or signaling through mechanoreceptors (e.g., platelet endothelial cell adhesion molecule (PECAM), vascular endothelial (VE)-cadherin, epithelial (E)-cadherin, integrin) embedded in the membrane. Certain mechanical stimulations through specific receptor complexes induce electrical and/or chemical impulses in cells and propagate across cells and tissues. These biomechanical sensory and biochemical responses have profound implications in normal physiology and disease. Here, we discuss the tools that facilitate the understanding of mechanosensitive adhesion receptors. This article is structured to provide a broad biochemical and mechanobiology background to introduce a freshman mechano-biologist to the field of mechanotransduction, with deeper study enabled by many of the references cited herein.

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Traction force microscopy in rapidly moving cells reveals separate roles for ROCK and MLCK in the mechanics of retraction

Cited by 13 publications

References 53 publications

Single cell force profiling of human myofibroblasts reveals a biophysical spectrum of cell states

Single cell force profiling of human myofibroblasts reveals a biophysical spectrum of cell states

Elevated BMP and Mechanical Signaling Through YAP1/RhoA Poises FOP Mesenchymal Progenitors for Osteogenesis

Biophysical Tools to Study Cellular Mechanotransduction

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