The Focal Adhesion-Localized CdGAP Regulates Matrix Rigidity Sensing and Durotaxis

Wormer, Duncan B.; Davis, Kevin; Henderson, James H.; Turner, Christopher E.

doi:10.1371/journal.pone.0091815

Cited by 53 publications

(66 citation statements)

References 70 publications

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“…23,24 The molecular connections between focal adhesions, cytoskeletal filaments and nuclear scaffolds may produce integrated changes in cellular and nuclear structure. 20,21,25 It has been shown that the differentiation of murine MSCs into a chondrocytic phenotype required a rounded cell shape.…”

Section: Introductionmentioning

confidence: 99%

Nanotopographical Modulation of Cell Function through Nuclear Deformation

Wang

Bruce

Mezan

et al. 2016

ACS Appl. Mater. Interfaces

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Although nanotopography has been shown to be a potent modulator of cell behavior, it is unclear how the nanotopographical cue, through focal adhesions, affects the nucleus, eventually influencing cell phenotype and function. Thus, current methods to apply nanotopography to regulate cell behavior are basically empirical. We, herein, engineered nanotopographies of various shapes (gratings and pillars) and dimensions (feature size, spacing and height), and thoroughly investigated cell spreading, focal adhesion organization and nuclear deformation of human primary fibroblasts as the model cell grown on the nanotopographies. We examined the correlation between nuclear deformation and cell functions such as cell proliferation, transfection and extracellular matrix protein type I collagen production. It was found that the nanoscale gratings and pillars could facilitate focal adhesion elongation by providing anchoring sites, and the nanogratings could orient focal adhesions and nuclei along the nanograting direction, depending on not only the feature size but also the spacing of the nanogratings. Compared with continuous nanogratings, discrete nanopillars tended to disrupt the formation and growth of focal adhesions and thus had less profound effects on nuclear deformation. Notably, nuclear volume could be effectively modulated by the height of nanotopography. Further, we demonstrated that cell proliferation, transfection, and type I collagen production were strongly associated with the nuclear volume, indicating that the nucleus serves as a critical mechanosensor for cell regulation. Our study delineated the relationships between focal adhesions, nucleus and cell function and highlighted that the nanotopography could regulate cell phenotype and function by modulating nuclear deformation. This study provides insight into the rational design of nanotopography for new biomaterials and the cell–substrate interfaces of implants and medical devices.

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Section: Introductionmentioning

confidence: 99%

Nanotopographical Modulation of Cell Function through Nuclear Deformation

Wang

Bruce

Mezan

et al. 2016

ACS Appl. Mater. Interfaces

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“…In particular, CdGAP (Cdc42 GTPase-activating protein)/ARHGAP31, a GAP for Rac1 and Cdc422526, was reported to be highly expressed in human umbilical vein endothelial cells (HUVECs)24, and to be regulated in Zebrafish by the transcription factor Ets, a master regulator of angiogenesis2728. Outside of the vascular system, pro-migratory and pro-invasive functions were ascribed to CdGAP, which was shown to regulate directional membrane protrusions of migrating osteosarcoma cells293031 and TGFβ-dependent cell motility and invasion of breast cancer cells32. Recently, truncating mutations in the terminal exon of the CdGAP gene were identified in patients with a rare developmental disorder, Adams-Oliver Syndrome (AOS)3334, which is frequently associated with cardiac and vascular anomalies3536.…”

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confidence: 99%

CdGAP/ARHGAP31, a Cdc42/Rac1 GTPase regulator, is critical for vascular development and VEGF-mediated angiogenesis

Caron

DeGeer

Fournier

et al. 2016

Sci Rep

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Mutations in the CdGAP/ARHGAP31 gene, which encodes a GTPase-activating protein for Rac1 and Cdc42, have been reported causative in the Adams-Oliver developmental syndrome often associated with vascular defects. However, despite its abundant expression in endothelial cells, CdGAP function in the vasculature remains unknown. Here, we show that vascular development is impaired in CdGAP-deficient mouse embryos at E15.5. This is associated with superficial vessel defects and subcutaneous edema, resulting in 44% embryonic/perinatal lethality. VEGF-driven angiogenesis is defective in CdGAP−/− mice, showing reduced capillary sprouting from aortic ring explants. Similarly, VEGF-dependent endothelial cell migration and capillary formation are inhibited upon CdGAP knockdown. Mechanistically, CdGAP associates with VEGF receptor-2 and controls VEGF-dependent signaling. Consequently, CdGAP depletion results in impaired VEGF-mediated Rac1 activation and reduced phosphorylation of critical intracellular mediators including Gab1, Akt, PLCγ and SHP2. These findings are the first to demonstrate the importance of CdGAP in embryonic vascular development and VEGF-induced signaling, and highlight CdGAP as a potential therapeutic target to treat pathological angiogenesis and vascular dysfunction.

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“…Importantly, unlike polyacrylamide substrates, the ECM components do not have to be chemically cross-linked to PDMS. Indeed, the fibronectin coating is approximately equivalent on both the PDMS and the glass surfaces, as determined by staining with an antibody specific to fibronectin 11 . Since previous studies have shown that cells behave the same on either PDMS or polyacrylamide substrates, the use of PDMS substrates provides a much easier model with which to study mechanotransduction in cells 10,12 .…”

Section: Discussionmentioning

confidence: 99%

“…We have recently used this assay to study how the focal adhesion signaling protein cdGAP regulates mechanosensing and durotaxis 11 . Clearly, there are numerous cellular components in addition to focal adhesions, such as cytoskeletal elements, protein trafficking components and ion channels that are involved in mechanical signaling and thus may also contribute to the regulation of durotaxis [13][14][15] .…”

Section: Discussionmentioning

confidence: 99%

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A Simplified System for Evaluating Cell Mechanosensing and Durotaxis <em>In Vitro</em>

Goreczny

Wormer

Turner

2015

JoVE

Self Cite

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The composition and mechanical properties of the extracellular matrix are highly variable between tissue types. This connective tissue stroma diversity greatly impacts cell behavior to regulate normal and pathologic processes including cell proliferation, differentiation, adhesion signaling and directional migration. In this regard, the innate ability of certain cell types to migrate towards a stiffer, or less compliant matrix substrate is referred to as durotaxis. This phenomenon plays an important role during embryonic development, wound repair and cancer cell invasion. Here, we describe a straightforward assay to study durotaxis, in vitro, using polydimethylsiloxane (PDMS) substrates. Preparation of the described durotaxis chambers creates a rigidity interface between the relatively soft PDMS gel and a rigid glass coverslip. In the example provided, we have used these durotaxis chambers to demonstrate a role for the cdc42/Rac1 GTPase activating protein, cdGAP, in mechanosensing and durotaxis regulation in human U2OS osteosarcoma cells. This assay is readily adaptable to other cell types and/or knockdown of other proteins of interests to explore their respective roles in mechanosignaling and durotaxis. Video LinkThe video component of this article can be found at

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The Focal Adhesion-Localized CdGAP Regulates Matrix Rigidity Sensing and Durotaxis

Cited by 53 publications

References 70 publications

Nanotopographical Modulation of Cell Function through Nuclear Deformation

Nanotopographical Modulation of Cell Function through Nuclear Deformation

CdGAP/ARHGAP31, a Cdc42/Rac1 GTPase regulator, is critical for vascular development and VEGF-mediated angiogenesis

A Simplified System for Evaluating Cell Mechanosensing and Durotaxis <em>In Vitro</em>

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