Growth control by intracellular tension and extracellular stiffness

doi:10.1016/j.tcb.2008.05.002

Trends in Cell Biology

Volume 18, Issue 7, July 2008, Pages 347-352

https://doi.org/10.1016/j.tcb.2008.05.002 Get rights and content

Integrin-mediated cell attachment to the extracellular matrix is an established regulator of the cell cycle, and the best-characterized targets of this process are the cyclin D1 gene and members of the cip and kip (cip/kip) family of cdk inhibitors. Manipulation of intracellular tension affects the same targets, supporting the idea that integrin activation and intracellular tension are closely related. Several signaling cascades, including FAK, Rho GTPases and ERK, transmit the integrin and tensional signals to pathways controlling the cell cycle. However, the experimental approaches that have generated these results alter cell adhesion and tension in ways that do not reflect the subtlety of those occurring in vivo. Increasing emphasis is therefore being placed on approaches that use micropatterning to control cell spreading, and deformable substrata to model the compliance of biological tissue.

Introduction

The extracellular microenvironment has profound effects on several cellular functions, including differentiation, apoptosis and proliferation. Although many studies have shown that localized release of soluble factors affects cellular function, the mixture of matrix proteins, proteoglycans and glucosaminoglycans that comprise the extracellular matrix (ECM) provide equally important cues that direct cellular decisions. Perhaps the newest idea about cellular control by the microenvironment is that the stiffness of the ECM (also referred to as its ‘compliance’) itself provides information to the cells. Several recent reviews have discussed the effects that ECM stiffness has on cell differentiation 1, 2. Here, we discuss data supporting the importance of the ECM, intracellular tension, cell shape, and ECM stiffness as regulators of integrin-dependent cell proliferation. Although we focus on studies showing that the ECM regulates progression through G₁ phase, we note a recent study indicating that integrin-mediated adhesion can also regulate M phase [3].

Section snippets

Regulation of cyclin D1 expression by the ECM

cyclin D1 has been termed a ‘mitogen sensor’ because its expression is induced by many mitogenic factors, including growth factors, cytokines and hormones. However, our early studies showed that the induction of cyclin D1 expression is blocked when mitogen-stimulated cells are unable to attach to the substratum (for review, see [4]). Some ECM components, such as fibronectin, synergize with growth factors to induce the expression of cyclin D1 mRNA, whereas others, such as hyaluronan [5] and

cyclin D1 gene expression requires ECM-mediated sustained ERK activity

Studies using pharmacological MAP-ERK kinase (MEK) inhibitors and dominant–negative or constitutively active mutants of the Extracellular signal-regulated protein kinase (ERK) cascade indicate that ERK activity – probably ERK5 in addition to the prototypical ERK1 and ERK2 [8] – stimulates cyclin D1 expression in mid-G₁ phase in many cell types (Figure 1; for review, see [9]). ERK activity stimulates the cyclin D1 promoter, whereas inhibition of ERK represses the cyclin D1 promoter;

FAK and cyclin D1 gene expression

Focal adhesion kinase (FAK) is the canonical mediator of integrin signals [11], and several reports have linked FAK to the induction of cyclin D1 (Figure 1). FAK is recruited to focal adhesions through its Focal adhesion targeting (FAT) domain and is autophosphorylated at Y397 upon ECM-induced integrin clustering. Phosphorylation of Y397 creates a binding site for Src and results in the Src-dependent tyrosine phosphorylation of FAK at several other sites, including Y576 and Y577–

ECM binding to integrins downregulates cip/kip cdk inhibitors

In addition to regulating expression of cyclin D1, the ECM regulates the expression of members of the cip/kip cdk inhibitor family (e.g. p21^cip1 and p27^kip1) which regulate activity of cdk2 in G₁ phase and entry into S phase (for review, see in 4, 16). Incubating cells in suspension blocks integrin signaling and leads to an increase in the levels of p21^cip1 and p27^kip1. Skp2 is the substrate-targeting component of the E3 ubiquitin ligase complex that targets p27^kip1 for degradation, and

Effects of intracellular tension, the actin cytoskeleton, and Rho family GTPases on adhesion-dependent signaling to the cell cycle

The actin cytoskeleton plays a major role in determining the degree of intracellular tension of a cell, and a high-tensional state is characterized by the appearance of actin stress fibers. Studies using actin-depolymerizing drugs have shown that sustained ERK activity and ERK-dependent cyclin D1 induction correlate with actin stress fiber formation and the consequent increase in intracellular tension (for review, see [4]). The Rho–Rho kinase pathway increases actin polymerization and cellular

Effects of cell spreading on integrin signaling and cell proliferation

Although the majority of studies of intracellular tension use either pharmacological inhibitors of actin polymerization or dominant–negative Rho GTPases to disrupt intracellular tension, an alternative approach is to control cell spreading. Cell spreading can be controlled by plating cells on different densities of matrix protein or by plating cells onto micropatterned, matrix protein-coated ‘islands’ of defined size; to date, most of the substrates have been rigid. Micropatterning enables

Modeling the effects of tissue and ECM compliance on integrin signaling and proliferation

Although actin-depolymerizing drugs and deliberate Rho–Rho kinase inhibition (e.g. with C3 toxin, dominant–negative constructs, RNA interference or Y27632) have been widely used to study the effects of integrin signaling on cellular function, these approaches probably result in much more severe changes in f-actin and Rho–Rho kinase activity than occur in vivo. Additionally, a pervasive shortcoming of these approaches is that the cells are cultured on non-deformable 2D substrata (i.e. culture

A working model for cell cycle control by intracellular tension and extracellular stiffness

Collectively, the results discussed above indicate that integrin-regulated cell cycle progression is not a binary process of ‘off’ or ‘on’ but rather a collection of signaling events with different compliance or tensional thresholds (Figure 2). For example, at the highest level of ECM compliance, FAK autophosphorylation is reduced, ERK activity is not sustained, cyclin D1 is not expressed, and cdk inhibitor expression is upregulated. This molecular signature, which results in G₁ phase arrest,

Conclusions and future directions

The extracellular matrix is remodeled physiologically and also pathologically in diseases as diverse as fibrosis, cancer and atherosclerosis. The results to date indicate that changes in ECM stiffness affect cell morphology, integrin signaling, and the actin cytoskeleton, thereby facilitating control of the cell cycle. Given that different integrin-dependent signaling events have distinct compliance thresholds, the molecular composition of integrin signaling complexes at high and low tissue

Acknowledgements

Work in our laboratory is supported by grants from the National Institutes of Health.

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Trends in Cell Biology

ReviewGrowth control by intracellular tension and extracellular stiffness

Introduction

Section snippets

Regulation of cyclin D1 expression by the ECM

cyclin D1 gene expression requires ECM-mediated sustained ERK activity

FAK and cyclin D1 gene expression

ECM binding to integrins downregulates cip/kip cdk inhibitors

Effects of intracellular tension, the actin cytoskeleton, and Rho family GTPases on adhesion-dependent signaling to the cell cycle

Effects of cell spreading on integrin signaling and cell proliferation

Modeling the effects of tissue and ECM compliance on integrin signaling and proliferation

A working model for cell cycle control by intracellular tension and extracellular stiffness

Conclusions and future directions

Acknowledgements

Curr. Opin. Genet. Dev.

Cancer Cell

Mol. Cell

J. Biol. Chem.

Neoplasia

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Exp. Cell Res.

Cancer Cell

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Cell

J. Biol. Chem.

Biomaterials

J. Biol. Chem.

Biophys. J.

Cell

Biophys. J.

Eur. J. Cell Biol.

Mater. Lett.

Tissue cells feel and respond to the stiffness of their substrate

Science

Cell type-specific response to growth on soft materials

J. Appl. Physiol.

Perturbing integrin function inhibits microtubule growth from centrosomes, spindle assembly, and cytokinesis

J. Cell Biol.

Hyaluronan and CD44 antagonize mitogen-dependent cyclin D1 expression in mesenchymal cells

J. Cell Biol.

SPARC regulates cell cycle progression in mesangial cells via its inhibition of IGF-dependent signaling

J. Cell. Biochem.

Glycogen synthase kinase-3beta regulates cyclin D1 proteolysis and subcellular localization

Genes Dev.

Activation of cyclin D1 expression by the ERK5 cascade

Oncogene

Integrating the MAP kinase signal into the G1 phase cell cycle machinery

Bioessays

ERK activity and G1 phase progression: identifying dispensable versus essential activities and primary versus secondary targets

Mol. Biol. Cell

Focal adhesion kinase: the first ten years

J. Cell Sci.

Integrin-mediated activation of focal adhesion kinase is required for signaling to Jun NH2-terminal kinase and progression through the G1 phase of the cell cycle

J. Cell Biol.

Transcriptional activation of cyclin D1 promoter by FAK contributes to cell cycle progression

Mol. Biol. Cell

Integrins and cell proliferation: regulation of cyclin-dependent kinases via cytoplasmic signaling pathways

J. Cell Sci.

Review
Growth control by intracellular tension and extracellular stiffness

Integrating the MAP kinase signal into the G₁ phase cell cycle machinery

ERK activity and G₁ phase progression: identifying dispensable versus essential activities and primary versus secondary targets

Integrin-mediated activation of focal adhesion kinase is required for signaling to Jun NH2-terminal kinase and progression through the G₁ phase of the cell cycle