Interplay of matrix stiffness and protein tethering in stem cell differentiation

Wen, Jessica H.; Vincent, Ludovic G.; Fuhrmann, Alexander; Choi, Yu Suk; Hribar, Kolin C.; Taylor‐Weiner, Hermes; Chen, Shaochen; Engler, Adam J.

doi:10.1038/nmat4051

Cited by 839 publications

(884 citation statements)

References 37 publications

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“…S2A), and thus differences in cell morphology might be due to differences in ligand density, rather than gel modulus. Previous studies demonstrated that protein tethering to the surface of PA gels can be modulated by varying the concentration of the substrate-protein cross-linker sulfo-SANPAH (SS) (41). Therefore, a range of SS concentrations (0.5-2.0 mg/mL) were used to functionalize a 60-kPa PA gel, and Ecad-Fc ligand binding was measured using an anti-E-cadherin antibody (Fig.…”

Section: Resultsmentioning

confidence: 99%

Changes in E-cadherin rigidity sensing regulate cell adhesion

Collins

Denisin

Pruitt

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Mechanical cues are sensed and transduced by cell adhesion complexes to regulate diverse cell behaviors. Extracellular matrix (ECM) rigidity sensing by integrin adhesions has been well studied, but rigidity sensing by cadherins during cell adhesion is largely unexplored. Using mechanically tunable polyacrylamide (PA) gels functionalized with the extracellular domain of E-cadherin (Ecad-Fc), we showed that E-cadherin-dependent epithelial cell adhesion was sensitive to changes in PA gel elastic modulus that produced striking differences in cell morphology, actin organization, and membrane dynamics. Traction force microscopy (TFM) revealed that cells produced the greatest tractions at the cell periphery, where distinct types of actin-based membrane protrusions formed. Cells responded to substrate rigidity by reorganizing the distribution and size of hightraction-stress regions at the cell periphery. Differences in adhesion and protrusion dynamics were mediated by balancing the activities of specific signaling molecules. Cell adhesion to a 30-kPa Ecad-Fc PA gel required Cdc42-and formin-dependent filopodia formation, whereas adhesion to a 60-kPa Ecad-Fc PA gel induced Arp2/3-dependent lamellipodial protrusions. A quantitative 3D cell-cell adhesion assay and live cell imaging of cell-cell contact formation revealed that inhibition of Cdc42, formin, and Arp2/3 activities blocked the initiation, but not the maintenance of established cell-cell adhesions. These results indicate that the same signaling molecules activated by E-cadherin rigidity sensing on PA gels contribute to actin organization and membrane dynamics during cell-cell adhesion. We hypothesize that a transition in the stiffness of E-cadherin homotypic interactions regulates actin and membrane dynamics during initial stages of cellcell adhesion.C ell adhesion is essential for tissue structure and function. Cells use specialized types of adhesions to interact with the surrounding environment, including integrin-based focal adhesions at cell-extracellular matrix (cell-ECM) contacts and cadherin-based adhesions at cell-cell contacts (1). Integrins bind to the ECM and intracellular proteins that link to the actin cytoskeleton and important signaling pathways (2). Similarly, cadherins regulate cellcell recognition and adhesion (3) and, through cytoplasmic adaptor proteins (catenins, vinculin) (4, 5), also link to the actin cytoskeleton and other proteins with signaling and scaffolding functions (6).Initiation of cell-cell adhesion requires significant reorganization of the actin cytoskeleton and is tightly controlled by the activities of actin nucleating proteins and Rho GTPases. Adhesion is initiated when filopodia from opposing cells come into contact with one another (7,8), and this process is regulated by Cdc42 activity (9, 10) and formin-dependent actin polymerization (11)(12)(13)(14). Intermediate stages of cell-cell contact formation involve lateral expansion of the contact by Rac1-induced and Arp2/3-dependent lamellipodial activity (15, 16). Finally, com...

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Section: Resultsmentioning

confidence: 99%

Changes in E-cadherin rigidity sensing regulate cell adhesion

Collins

Denisin

Pruitt

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…Our amyloid-based hydrogel system promotes the commitment of stem cells toward neurons, and this effect may be mediated by mechanotransduction. [46][47][48] Hence, this type of hydrogel can promote neuronal differentiation without the need for any additional growth factors. Acute inflammation followed by immune rejection is often associated with many scaffolds implanted in the brain, making them unsuitable for in vivo applications.…”

Section: Discussionmentioning

confidence: 99%

Implantable amyloid hydrogels for promoting stem cell differentiation to neurons

et al. 2016

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We report a new class of amyloid-inspired peptide hydrogels that was designed and based on α-synuclein protein for which hydrogel formation is triggered by various stimuli, such as heating/cooling or changes in pH. The peptides resemble a cross-β-sheet-rich amyloid, and they assemble into a nanofibrous meshwork that mimics the natural extracellular matrix. Our design principle allows easy manipulation of the gelator sequence to exploit the desirable properties of amyloids for use in cell replacement therapies for neurodegenerative diseases. The amyloid hydrogels facilitate the attachment and neuronal differentiation of mesenchymal stem cells (MSCs) and assist in the delivery and engraftment of MSCs in the substantia nigra and caudate putamen of a Parkinsonian mouse model.

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“…As our data demonstrates a strong effect of composition on structural and mechanical (bulk rheological) properties, these may be more important to consider for the optimization of fibrin hydrogels for a specific application. Measured storage moduli covered two orders of magnitude within a range that is known to affect cell behavior in terms of differentiation [10,65], and also independently from the material porosity [66]. Low fibrinogen concentrations with relatively low thrombin and factor XIII concentrations (such as 5LL…”

Section: Discussionmentioning

confidence: 94%