Enhancement of endothelialization by topographical features is mediated by PTP1B-dependent endothelial adherens junctions remodeling

RationaleFailure of small synthetic vascular grafts is largely due to late endothelialization and has been an ongoing challenge in the treatment of cardiovascular diseases.ObjectivePrevious strategies developed to promote graft endothelialization include surface topographical modulation and biochemical modifications. However, these have been met with limited success. Importantly, although the integrity of Endothelial Cell (EC) monolayer is crucial for endothelialization, the crosstalk between surface topography and cell-cell connectivity is still not well understood. Here we explored a combined strategy that utilizes both topographical features and pharmacological perturbations.Methods and resultWe characterized EC behaviors in response to micron-scale grating topography in conjunction with pharmacological perturbations of endothelial adherens junctions (EAJ) regulators. We studied the EA.hy 926 cell-cell junctions and monolayer integrity using the junctional markers upon the inhibitory effect of EAJ regulator on both planar and grating topographies substrates.We identified a protein tyrosine phosphatase, PTP1B, as a potent regulator of EAJ stability. Next, we studied the physiologically relevant behaviors of EC using primary human coronary arterial endothelial cells (HCAEC). Our results showed that PTP1B inhibition synergized with grating topographies to modulate EAJ rearrangement, thereby controlling global EC monolayer sheet orientation, connectivity and collective cell migration to promote endothelialization.Our results showed that PTP1B inhibition synergized with grating topographies to modulate EAJ rearrangement, thereby controlling global EC monolayer sheet orientation, connectivity and collective cell migration and proliferation.ConclusionThe synergistic effect of PTP1B inhibition and grating topographies could be useful for the promotion of endothelialization by enhancing EC migration and proliferation.

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Faculty Opinions recommendation of α-Catenin Controls the Anisotropy of Force Distribution at Cell-Cell Junctions during Collective Cell Migration.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.733465778.793561842 ◽

2019 ◽

Author(s):

Johan De Rooij

Keyword(s):

Cell Migration ◽

Cell Junctions ◽

Force Distribution ◽

Collective Cell Migration ◽

Cell Cell

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HAX1 impact on collective cell migration, cell adhesion, and cell shape is linked to the regulation of actomyosin contractility

Molecular Biology of the Cell ◽

10.1091/mbc.e19-05-0304 ◽

2019 ◽

Vol 30 (25) ◽

pp. 3024-3036 ◽

Cited By ~ 3

Author(s):

Anna Balcerak ◽

Alicja Trebinska-Stryjewska ◽

Maciej Wakula ◽

Mateusz Chmielarczyk ◽

Urszula Smietanka ◽

...

Keyword(s):

Breast Cancer ◽

Cell Migration ◽

Single Cell ◽

Cancer Progression ◽

Cancer Cell Line ◽

Cell Junctions ◽

Collective Cell Migration ◽

Actomyosin Contractility ◽

Epithelial Cell Layer ◽

Cell Cell

HAX1 protein is involved in the regulation of apoptosis, cell motility and calcium homeostasis. Its overexpression was reported in several tumors, including breast cancer. This study demonstrates that HAX1 has an impact on collective, but not single-cell migration, thus indicating the importance of cell–cell contacts for the HAX1-mediated effect. Accordingly, it was shown that HAX1 knockdown affects cell–cell junctions, substrate adhesion, and epithelial cell layer integrity. As demonstrated here, these effects can be attributed to the modulation of actomyosin contractility through changes in RhoA and septin signaling. Additionally, it was shown that HAX1 does not influence invasive potential in the breast cancer cell line, suggesting that its role in breast cancer progression may be linked instead to collective invasion of the epithelial cells but not single-cell dissemination.

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The tyrosine phosphatase SHP2 regulates recovery of endothelial adherens junctions through control of β-catenin phosphorylation

Molecular Biology of the Cell ◽

10.1091/mbc.e12-01-0038 ◽

2012 ◽

Vol 23 (21) ◽

pp. 4212-4225 ◽

Cited By ~ 56

Author(s):

Ilse Timmerman ◽

Mark Hoogenboezem ◽

Anton M. Bennett ◽

Dirk Geerts ◽

Peter L. Hordijk ◽

...

Keyword(s):

Barrier Function ◽

Adherens Junctions ◽

Tyrosine Phosphatase ◽

Endothelial Permeability ◽

Cell Junctions ◽

Endothelial Barrier ◽

Endothelial Barrier Function ◽

Src Homology 2 Domain ◽

Cell Cell

Impaired endothelial barrier function results in a persistent increase in endothelial permeability and vascular leakage. Repair of a dysfunctional endothelial barrier requires controlled restoration of adherens junctions, comprising vascular endothelial (VE)-cadherin and associated β-, γ-, α-, and p120-catenins. Little is known about the mechanisms by which recovery of VE-cadherin–mediated cell–cell junctions is regulated. Using the inflammatory mediator thrombin, we demonstrate an important role for the Src homology 2-domain containing tyrosine phosphatase (SHP2) in mediating recovery of the VE-cadherin–controlled endothelial barrier. Using SHP2 substrate-trapping mutants and an in vitro phosphatase activity assay, we validate β-catenin as a bona fide SHP2 substrate. SHP2 silencing and SHP2 inhibition both result in delayed recovery of endothelial barrier function after thrombin stimulation. Moreover, on thrombin challenge, we find prolonged elevation in tyrosine phosphorylation levels of VE-cadherin–associated β-catenin in SHP2-depleted cells. No disassembly of the VE-cadherin complex is observed throughout the thrombin response. Using fluorescence recovery after photobleaching, we show that loss of SHP2 reduces the mobility of VE-cadherin at recovered cell–cell junctions. In conclusion, our data show that the SHP2 phosphatase plays an important role in the recovery of disrupted endothelial cell–cell junctions by dephosphorylating VE-cadherin–associated β-catenin and promoting the mobility of VE-cadherin at the plasma membrane.

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A-CAM: a 135-kD receptor of intercellular adherens junctions. II. Antibody-mediated modulation of junction formation.

The Journal of Cell Biology ◽

10.1083/jcb.103.4.1451 ◽

1986 ◽

Vol 103 (4) ◽

pp. 1451-1464 ◽

Cited By ~ 84

Author(s):

T Volk ◽

B Geiger

Keyword(s):

Cell Adhesion ◽

Adherens Junctions ◽

Adherens Junction ◽

Recovery Phase ◽

Inhibitory Effect ◽

Cell Junctions ◽

Specific Cell ◽

Normal Medium ◽

Junction Formation ◽

Cell Cell

Intercellular adherens junctions between cultured lens epithelial cells are highly Ca2+-dependent and are readily dissociated upon chelation of extracellular Ca2+ ions. Addition of Ca2+ to EGTA-treated cells results in the recovery of cell-cell junctions including the reorganization of adherens junction-specific cell adhesion molecule (A-CAM), vinculin, and actin (Volk, T., and B. Geiger, 1986, J. Cell Biol., 103:000-000). Incubation of cells during the recovery phase with Fab' fragments of anti-A-CAM specifically inhibited the re-formation of cell-cell adherens junctions. This inhibition was accompanied by remarkable changes in microfilament organization manifested by an apparent deterioration of stress fibers and the appearance of fragmented actin bundles throughout the cytoplasm. Incubation of EGTA-dissociated cells with intact divalent anti-A-CAM antibodies in normal medium had no apparent inhibitory effect on junction formation and did not affect the assembly of actin microfilament bundles. Moreover, adherens junctions formed in the presence of the divalent antibodies became essentially Ca2+-independent, suggesting that cell-cell adhesion between them was primarily mediated by the antibodies. These studies suggest that A-CAM participates in intercellular adhesion in adherens-type junctions and point to its involvement in microfilament bundle assembly.

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Protein Phosphatase 1 activity controls a balance between collective and single cell modes of migration

10.1101/811562 ◽

2019 ◽

Author(s):

Yujun Chen ◽

Nirupama Kotian ◽

George Aranjuez ◽

Lin Chen ◽

C. Luke Messer ◽

...

Keyword(s):

Cell Migration ◽

Single Cell ◽

Protein Phosphatase ◽

Single Cells ◽

Cell Junctions ◽

Protein Phosphatase 1 ◽

Collective Cell Migration ◽

Border Cells ◽

Border Cell ◽

Cell Cell

AbstractCollective cell migration is central to many developmental and pathological processes. However, the mechanisms that keep cell collectives together and coordinate movement of multiple cells are poorly understood. Using the Drosophila border cell migration model, we find that Protein phosphatase 1 (Pp1) activity controls collective cell cohesion and migration. Inhibition of Pp1 causes border cells to round up, dissociate, and move as single cells with altered motility. We present evidence that Pp1 promotes proper levels of cadherin-catenin complex proteins at cell-cell junctions within the cluster to keep border cells together. Pp1 further restricts actomyosin contractility to the cluster periphery rather than at internal cell-cell contacts. We show that the myosin phosphatase Pp1 complex, which inhibits non-muscle myosin-II (Myo-II) activity, coordinates border cell shape and cluster cohesion. Given the high conservation of Pp1 complexes, this study identifies Pp1 as a major regulator of collective versus single cell migration.

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Short-term stimulation of collective cell migration in tissues reprograms long-term supracellular dynamics

10.1101/2021.07.27.453602 ◽

2021 ◽

Author(s):

Abraham E. Wolf ◽

Matthew A. Heinrich ◽

Isaac B. Breinyn ◽

Tom J. Zajdel ◽

Daniel J. Cohen

Keyword(s):

Cell Migration ◽

Invasive Disease ◽

Cell Junctions ◽

Collective Cell Migration ◽

Long Term Effects ◽

Short Term ◽

Cell Dynamics ◽

Global Migration ◽

Cell Cell

The ability to program collective cell migration can allow us to control critical multicellular processes in development, regenerative medicine, and invasive disease. However, while various technologies exist to make individual cells migrate, translating these tools to control myriad, collectively interacting cells within a single tissue poses many challenges. For instance, do cells within the same tissue interpret a global migration 'command' differently based on where they are in the tissue? Similarly, since no stimulus is permanent, what are the long-term effects of transient commands on collective cell dynamics? We investigate these questions by bioelectrically programming large epithelial tissues to globally migrate 'rightward' via electrotaxis. Tissues clearly developed distinct rear, middle, side, and front responses to a single global migration stimulus. Furthermore, at no point post-stimulation did tissues return to their pre-stimulation behavior, instead equilibrating to a third, new migratory state. These unique dynamics suggested that programmed migration resets tissue mechanical state, which was confirmed by transient chemical disruption of cell-cell junctions, analysis of strain wave propagation patterns, and quantification of cellular crowd dynamics. Overall, this work demonstrates how externally driving the collective migration of a tissue can reprogram baseline cell-cell interactions and collective dynamics, even well beyond the end of the global migratory cue, and emphasizes the importance of considering the supracellular context of tissues and other collectives when attempting to program crowd behaviors.

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Overriding native cell coordination enhances external programming of collective cell migration

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2101352118 ◽

2021 ◽

Vol 118 (29) ◽

pp. e2101352118

Author(s):

Gawoon Shim ◽

Danelle Devenport ◽

Daniel J. Cohen

Keyword(s):

Cell Migration ◽

Cell Adhesion ◽

Mouse Skin ◽

Cell Junctions ◽

External Control ◽

Collective Cell Migration ◽

Collective Behaviors ◽

Primary Mouse ◽

E Cadherin ◽

Cell Cell

As collective cell migration is essential in biological processes spanning development, healing, and cancer progression, methods to externally program cell migration are of great value. However, problems can arise if the external commands compete with strong, preexisting collective behaviors in the tissue or system. We investigate this problem by applying a potent external migratory cue—electrical stimulation and electrotaxis—to primary mouse skin monolayers where we can tune cell–cell adhesion strength to modulate endogenous collectivity. Monolayers with high cell–cell adhesion showed strong natural coordination and resisted electrotactic control, with this conflict actively damaging the leading edge of the tissue. However, reducing preexisting coordination in the tissue by specifically inhibiting E-cadherin–dependent cell–cell adhesion, either by disrupting the formation of cell–cell junctions with E-cadherin–specific antibodies or rapidly dismantling E-cadherin junctions with calcium chelators, significantly improved controllability. Finally, we applied this paradigm of weakening existing coordination to improve control and demonstrate accelerated wound closure in vitro. These results are in keeping with those from diverse, noncellular systems and confirm that endogenous collectivity should be considered as a key quantitative design variable when optimizing external control of collective migration.

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ERK-mediated mechanochemical waves direct collective cell polarization

10.1101/2019.12.25.888552 ◽

2019 ◽

Cited By ~ 6

Author(s):

Naoya Hino ◽

Leone Rossetti ◽

Ariadna Marín-Llauradó ◽

Kazuhiro Aoki ◽

Xavier Trepat ◽

...

Keyword(s):

Cell Migration ◽

Cell Polarization ◽

Cell Junctions ◽

Collective Cell Migration ◽

Mechanical Forces ◽

Force Transmission ◽

Long Distance ◽

Erk Activation ◽

Guidance Cues ◽

Cell Cell

AbstractDuring collective migration of epithelial cells, the migration direction is aligned over a large, tissue-scale expanse. Although the collective cell migration is known to be directed by mechanical forces transmitted via cell-cell junctions, it remains elusive how the intercellular force transmission is coordinated with intracellular biochemical signaling to achieve collective movements. Here we show that intercellular coupling of extracellular signal-regulated kinase (ERK)-mediated mechanochemical feedback in individual cells yields long-distance transmission of guidance cues. Mechanical stretch activates ERK through epidermal growth factor receptor (EGFR) activation, and the ERK activation triggers cell contraction. In addition, the contraction of the activated cell pulls neighboring cells via cell-cell junctions, evoking another round of ERK activation and contraction in the neighbors. Furthermore, anisotropic contraction based on front-rear cell polarization guarantees unidirectional propagation of ERK activation waves, and in turn, the ERK activation waves direct multicellular alignment of the polarity, leading to long-range ordered migration. Our findings reveal that mechanical forces mediate intercellular signaling underlying sustained transmission of guidance cues for collective cell migration.

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