A new micropatterning method of soft substrates reveals that different tumorigenic signals can promote or reduce cell contraction levels

Tseng, Qingzong; Wang, Irène; Duchemin-Pelletier, Eve; Azioune, Ammar; Carpi, Nicolas; Gao, Jie; Filhol, Odile; Piel, Matthieu; Théry, Manuel; Balland, Martial

doi:10.1039/c0lc00641f

Cited by 223 publications

(230 citation statements)

References 53 publications

(69 reference statements)

Supporting

Mentioning

211

Contrasting

Order By: Relevance

“…Addition of the oxidized protein to the hydrogel surface leads to rapid conjugation within 1 h. To verify bioconjugation, we first mixed an Alexa-647-conjugated fibrinogen with the selected matrix protein to confirm protein conjugation and pattern fidelity as demonstrated in a previous report. 32 Immunofluorescence analysis of the conjugated gels indicates higher fluorescence intensity on the gels that were treated with hydrazine, thus confirming conjugation (Fig. 1C).…”

Section: Polyacrylamide Gel Fabrication and Chemical Modificationsupporting

confidence: 49%

Matrix Composition and Mechanics Direct Proangiogenic Signaling from Mesenchymal Stem Cells

Abdeen

Weiss

Lee

et al. 2014

Tissue Engineering Part A

104

122

View full text Add to dashboard Cite

The secretion of trophic factors that promote angiogenesis from mesenchymal stem cells (MSCs) is a promising cell-based therapeutic treatment. However, clinical efficacy has proved variable, likely on account of ill-defined cell delivery formulations and the inherent complexity of cellular secretion. Here we show how controlling the mechanical properties and protein composition of the extracellular matrix (ECM) surrounding MSCs can guide proangiogenic signaling. Conditioned media from MSCs adherent to polyacrylamide hydrogel functionalized with fibronectin, collagen I, or laminin was applied to 3D matrigel cultures containing human microvascular endothelial cells (HMVECs). The degree of tubulogenesis in HMVECs is shown to depend on both the substrate rigidity and matrix protein composition. MSCs cultured on fibronectin-modified hydrogels show a stiffness dependence in proangiogenic signaling with maximum influence on tubulogenesis observed from 40 kPa conditioned media, twofold higher than commercially available cocktails of growth factors. Quantitative real-timepolymerase chain reaction reveals stiffness-dependent expression of multiple factors involved in angiogenesis that corroborate the functional tubulogenesis assay. Restricting cell spreading with micropatterned surfaces attenuates the conditioned media effects; however, small-molecule inhibitors of actomyosin contractility do not significantly reduce the functional outcome. This work demonstrates how controlling matrix rigidity and protein composition can influence the secretory profile of MSCs. Model systems that deconstruct the physical and biochemical cues involved in MSC secretion may assist in the design of hydrogel biomaterials for cell-based therapies.

show abstract

Section: Polyacrylamide Gel Fabrication and Chemical Modificationsupporting

confidence: 49%

Matrix Composition and Mechanics Direct Proangiogenic Signaling from Mesenchymal Stem Cells

Abdeen

Weiss

Lee

et al. 2014

Tissue Engineering Part A

104

122

View full text Add to dashboard Cite

show abstract

“…Cells were plated at an approximate density of 2×10 4 cells per cm 2 for 3-4 h and images were acquired on an iMIC Andromeda (FEI, Gräfelfing, Germany) microscope at 40x magnification. Force calculations were performed as previously described (Tseng et al, 2011).…”

Section: Traction Force Microscopymentioning

confidence: 99%

ICAP-1 monoubiquitylation coordinates matrix density and rigidity sensing for cell migration through ROCK2–MRCKα balance

Bouin

Kyumurkov

Régent-Kloeckner

et al. 2017

Journal of Cell Science

Self Cite

View full text Add to dashboard Cite

There was an error published in J. Cell Sci. 130, 626-636.The name of the author Alexander Kyumurkov was incorrect in the original author list. The correct spelling is given above.The authors apologise to the readers for any confusion that this error might have caused. 1195 ABSTRACTCell migration is a complex process requiring density and rigidity sensing of the microenvironment to adapt cell migratory speed through focal adhesion and actin cytoskeleton regulation. ICAP-1 (also known as ITGB1BP1), a β1 integrin partner, is essential for ensuring integrin activation cycle and focal adhesion formation. We show that ICAP-1 is monoubiquitylated by Smurf1, preventing ICAP-1 binding to β1 integrin. The non-ubiquitylatable form of ICAP-1 modifies β1 integrin focal adhesion organization and interferes with fibronectin density sensing. ICAP-1 is also required for adapting cell migration in response to substrate stiffness in a β1-integrin-independent manner. ICAP-1 monoubiquitylation regulates rigidity sensing by increasing MRCKα (also known as CDC42BPA)-dependent cell contractility through myosin phosphorylation independently of substrate rigidity. We provide evidence that ICAP-1 monoubiquitylation helps in switching from ROCK2-mediated to MRCKα-mediated cell contractility. ICAP-1 monoubiquitylation serves as a molecular switch to coordinate extracellular matrix density and rigidity sensing thus acting as a crucial modulator of cell migration and mechanosensing.

show abstract

“…Traction force microscopy calculations were performed as previously described (48). A description may also be found in SI Materials and Methods.…”

Section: Methodsmentioning

confidence: 99%

RhoA knockout fibroblasts lose tumor-inhibitory capacity in vitro and promote tumor growth in vivo

Alkasalias

Alexeyenko

Hennig

et al. 2017

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

Self Cite

View full text Add to dashboard Cite

Fibroblasts are a main player in the tumor-inhibitory microenvironment. Upon tumor initiation and progression, fibroblasts can lose their tumor-inhibitory capacity and promote tumor growth. The molecular mechanisms that underlie this switch have not been defined completely. Previously, we identified four proteins overexpressed in cancer-associated fibroblasts and linked to Rho GTPase signaling. Here, we show that knocking out the Ras homolog family member A (RhoA) gene in normal fibroblasts decreased their tumorinhibitory capacity, as judged by neighbor suppression in vitro and accompanied by promotion of tumor growth in vivo. This also induced PC3 cancer cell motility and increased colony size in 2D cultures. RhoA knockout in fibroblasts induced vimentin intermediate filament reorganization, accompanied by reduced contractile force and increased stiffness of cells. There was also loss of wide F-actin stress fibers and large focal adhesions. In addition, we observed a significant loss of α-smooth muscle actin, which indicates a difference between RhoA knockout fibroblasts and classic cancer-associated fibroblasts. In 3D collagen matrix, RhoA knockout reduced fibroblast branching and meshwork formation and resulted in more compactly clustered tumor-cell colonies in coculture with PC3 cells, which might boost tumor stem-like properties. Coculturing RhoA knockout fibroblasts and PC3 cells induced expression of proinflammatory genes in both. Inflammatory mediators may induce tumor cell stemness. Network enrichment analysis of transcriptomic changes, however, revealed that the Rho signaling pathway per se was significantly triggered only after coculturing with tumor cells. Taken together, our findings in vivo and in vitro indicate that Rho signaling governs the inhibitory effects by fibroblasts on tumor-cell growth.Rho GTPases | RhoA | cancer-associated fibroblasts | tumor-inhibitory capacity | cytoskeleton

show abstract

A new micropatterning method of soft substrates reveals that different tumorigenic signals can promote or reduce cell contraction levels

Cited by 223 publications

References 53 publications

Matrix Composition and Mechanics Direct Proangiogenic Signaling from Mesenchymal Stem Cells

Matrix Composition and Mechanics Direct Proangiogenic Signaling from Mesenchymal Stem Cells

ICAP-1 monoubiquitylation coordinates matrix density and rigidity sensing for cell migration through ROCK2–MRCKα balance

RhoA knockout fibroblasts lose tumor-inhibitory capacity in vitro and promote tumor growth in vivo

Contact Info

Product

Resources

About