Rho kinase modulates the actin cytoskeleton in intestinal epithelial cells and is essential for epithelial cell migration during wound healing

Polyamines are essential for early mucosal restitution that occurs by epithelial cell migration to reseal superficial wounds after injury. Normal intestinal epithelial cells are tightly bound in sheets, but they need to be rapidly disassembled during restitution. β-Catenin is involved in cell-cell adhesion, and its tyrosine phosphorylation causes disassembly of adhesion junctions, enhancing the spreading of cells. The current study determined whether polyamines are required for the stimulation of epithelial cell migration by altering β-catenin tyrosine phosphorylation. Migration of intestinal epithelial cells (IEC-6 line) after wounding was associated with an increase in β-catenin tyrosine phosphorylation, which decreased the binding activity of β-catenin to α-catenin. Polyamine depletion by α-difluoromethylornithine reduced cytoplasmic free Ca2+concentration ([Ca2+]cyt), prevented induction of β-catenin phosphorylation, and decreased cell migration. Elevation of [Ca2+]cyt induced by the Ca2+ ionophore ionomycin restored β-catenin phosphorylation and promoted migration in polyamine-deficient cells. Decreased β-catenin phosphorylation through the tyrosine kinase inhibitor herbimycin-A or genistein blocked cell migration, which was accompanied by reorganization of cytoskeletal proteins. These results indicate that β-catenin tyrosine phosphorylation plays a critical role in polyamine-dependent cell migration and that polyamines induce β-catenin tyrosine phosphorylation at least partially through [Ca2+]cyt.

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CXCL12 activation of CXCR4 regulates mucosal host defense through stimulation of epithelial cell migration and promotion of intestinal barrier integrity

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00208.2004 ◽

2005 ◽

Vol 288 (2) ◽

pp. G316-G326 ◽

Cited By ~ 50

Author(s):

Jennifer M. Smith ◽

Priscilla A. Johanesen ◽

Michael K. Wendt ◽

David G. Binion ◽

Michael B. Dwinell

Keyword(s):

Wound Healing ◽

Cell Migration ◽

Epithelial Cell ◽

Host Defense ◽

Intestinal Epithelial Cell ◽

Mucosal Barrier ◽

Intestinal Epithelial ◽

Barrier Integrity ◽

Epithelial Migration ◽

Epithelial Cell Migration

Intestinal epithelial cell migration plays a key role in gastrointestinal mucosal barrier formation, enterocyte development, differentiation, turnover, wound healing, and adenocarcinoma metastasis. Chemokines, through engagement of their corresponding receptors, are potent mediators of directed cell migration and are critical in the establishment and regulation of innate and adaptive immune responses. The aim of this study was to define the role for the chemokine CXCL12 and its sole cognate receptor CXCR4 in regulating intestinal epithelial cell migration and to determine its impact on barrier integrity. CXCL12 stimulated the dose-dependent chemotactic migration of human T84 colonic epithelial cells. Epithelial cell migration was inhibited by CXCR4 neutralizing antibody, pertussis toxin, LY-294002, and PD-98059, thereby implicating Gαi, phosphatidylinositol 3-kinase (PI3-kinase), and the ERK1/2 MAP kinase pathways in CXCR4-specific signaling. CXCL12 was also shown to increase barrier integrity, as defined by transepithelial resistance and paracellular flux across differentiating T84 monolayers. To determine whether CXCL12 regulated epithelial restitution, we used the normal nontransformed intestinal epithelial cell-6 (IEC-6) wound healing model. By using RT-PCR, immunoblot analysis, and immunofluorescence microscopy, we first showed expression of both CXCR4 and its ligand by IEC-6 cells. We then demonstrated that CXCL12 activated comparable signaling mechanisms to stimulate epithelial migration in the absence of proliferation in wounded IEC-6 monolayers. Taken together, these data indicate that CXCL12 signaling via CXCR4 directs intestinal epithelial cell migration, barrier maturation, and restitution, consistent with an important mechanistic role for these molecules in mucosal barrier integrity and innate host defense.

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Tumor necrosis factor regulates intestinal epithelial cell migration by receptor-dependent mechanisms

AJP Cell Physiology ◽

10.1152/ajpcell.00309.2002 ◽

2003 ◽

Vol 284 (4) ◽

pp. C953-C961 ◽

Cited By ~ 81

Author(s):

Julissa Corredor ◽

Fang Yan ◽

Christopher C. Shen ◽

Wei Tong ◽

Sutha K. John ◽

...

Keyword(s):

Cell Migration ◽

Tumor Necrosis Factor ◽

Epithelial Cell ◽

Tumor Necrosis ◽

Intestinal Epithelial Cell ◽

Wound Closure ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Epithelial Cell Migration ◽

Necrosis Factor

Altered mucosal integrity and increased cytokine production, including tumor necrosis factor (TNF), are the hallmarks of inflammatory bowel disease (IBD). In this study, we addressed the role of TNF receptors (TNFR) on intestinal epithelial cell migration in an in vitro wound closure model. With mouse TNFR1 or TNFR2 knockout intestinal epithelial cells, gene transfection, and pharmacological inhibitors, we show a concentration-dependent receptor-mediated regulation of intestinal cell migration by TNF. A physiological TNF level (1 ng/ml) enhances migration through TNFR2, whereas a pathological level (100 ng/ml) inhibits wound closure through TNFR1. Increased rate of wound closure by TNFR2 or inhibition by TNFR1 cannot be explained by either increased proliferation or apoptosis, respectively. Furthermore, inhibiting Src tyrosine kinase decreases TNF-induced focal adhesion kinase (FAK) tyrosine phosphorylation and cellular migration. We therefore conclude that TNFR2 activates a novel Src-regulated pathway involving FAK tyrosine phosphorylation that enhances migration of intestinal epithelial cells.

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Resolvin E1 is a pro-repair molecule that promotes intestinal epithelial wound healing

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1921335117 ◽

2020 ◽

Vol 117 (17) ◽

pp. 9477-9482 ◽

Cited By ~ 5

Author(s):

Miguel Quiros ◽

Darius Feier ◽

Dorothee Birkl ◽

Rachit Agarwal ◽

Dennis W. Zhou ◽

...

Keyword(s):

Wound Healing ◽

Cell Migration ◽

Epithelial Cell ◽

Wound Repair ◽

Intestinal Epithelial Cell ◽

Intestinal Inflammation ◽

Small Gtpase ◽

Intestinal Epithelial ◽

Epithelial Cell Migration ◽

Cell Migration And Proliferation

Resolution of intestinal inflammation and wound repair are active processes that mediate epithelial healing at mucosal surfaces. Lipid molecules referred to as specialized proresolving mediators (SPMs) play an important role in the restorative response. Resolvin E1 (RvE1), a SPM derived from omega-3 fatty acids, has been reported to dampen intestinal inflammation by promoting anti-inflammatory responses including increased neutrophil spherocytosis and macrophage production of IL-10. Despite these observations, a role for RvE1 in regulating intestinal epithelial cell migration and proliferation during mucosal wound repair has not been explored. Using an endoscopic biopsy-based wound healing model, we report that RvE1 is locally produced in response to intestinal mucosal injury. Exposure of intestinal epithelial cells to RvE1 promoted wound repair by increasing cellular proliferation and migration through activation of signaling pathways including CREB, mTOR, and Src-FAK. Additionally, RvE1-triggered activation of the small GTPase Rac1 led to increased intracellular reactive oxygen species (ROS) production, cell–matrix adhesion, and cellular protrusions at the leading edge of migrating cells. Furthermore, in situ administration of RvE1-encapsulated synthetic targeted polymeric nanoparticles into intestinal wounds promoted mucosal repair. Together, these findings demonstrate that RvE1 functions as a prorepair lipid mediator by increasing intestinal epithelial cell migration and proliferation, and highlight potential therapeutic applications for this SPM to promote mucosal healing in the intestine.

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ADAM-15 inhibits wound healing in human intestinal epithelial cell monolayers

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00262.2004 ◽

2005 ◽

Vol 288 (2) ◽

pp. G346-G353 ◽

Cited By ~ 35

Author(s):

Laetitia Charrier ◽

Yutao Yan ◽

Adel Driss ◽

Christian L. Laboisse ◽

Shanthi V. Sitaraman ◽

...

Keyword(s):

Wound Healing ◽

Epithelial Cell ◽

Epithelial Cells ◽

Intestinal Epithelial Cell ◽

Intestinal Epithelial Cells ◽

Epithelial Cell Line ◽

Intestinal Epithelial ◽

Coding Region ◽

Human Intestinal Epithelial Cell ◽

New Variant

The disintegrin metalloproteases (or ADAMs) are membrane-anchored glycoproteins that have been implicated in cell-cell or cell-matrix interactions and in proteolysis of molecules on the cell surface. The expression and/or the pathophysiological implications of ADAMs are not known in intestinal epithelial cells. Therefore, our aim was to investigate the expression and the role of ADAMs in intestinal epithelial cells. Expression of ADAMs was assessed by RT-PCR, Western blot analysis, and immunufluorescence experiments. Wound-healing experiments were performed by using the electric cell substrate impedence sensing technology. Our results showed that ADAMs-10, -12, and -15 mRNA are expressed in the colonic human cell lines Caco2-BBE and HT29-Cl.19A. An ADAM-15 complementary DNA cloned from Caco2-BBE poly(A)+ RNA, and encompassing the entire coding region, was found to be shorter and to present a different region encoding the cytoplasmic tail compared with ADAM-15 sequence deposited in the database. In Caco2-BBE cells and colonic epithelial cells, ADAM-15 protein was found in the apical, basolateral, and intracellular compartments. We also showed that the overexpression of ADAM-15 reduced cell migration in a wound-healing assay in Caco2-BBE monolayers. Our data show that 1) ADAM-15 is expressed in human intestinal epithelia, 2) a new variant of ADAM-15 is expressed in a human intestinal epithelial cell line, and 3) ADAM-15 is involved in intestinal epithelial cells wound-healing processes. Together, these results suggest that ADAM-15 may have important pathophysiological roles in intestinal cells.

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Desmocollin-2 promotes intestinal mucosal repair by controlling integrin-dependent cell adhesion and migration

Molecular Biology of the Cell ◽

10.1091/mbc.e19-12-0692 ◽

2020 ◽

Vol 31 (6) ◽

pp. 407-418 ◽

Cited By ~ 2

Author(s):

Sven Flemming ◽

Anny-Claude Luissint ◽

Dennis H. M. Kusters ◽

Arturo Raya-Sandino ◽

Shuling Fan ◽

...

Keyword(s):

Wound Healing ◽

Epithelial Cell ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Epithelial Cell Migration ◽

Cell Adhesion And Migration ◽

Dependent Cell ◽

And Migration ◽

Mucosal Wound

We report a key role for Dsc2 in simple epithelial cell migration and mucosal wound healing in vivo using newly generated mice with inducible conditional knockdown of Dsc2 in intestinal epithelial cells ( Villin-CreERT2; Dsc2fl/fl).

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Proliferation, not apoptosis, alters epithelial cell migration in small intestine of CFTR null mice

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.2001.281.3.g681 ◽

2001 ◽

Vol 281 (3) ◽

pp. G681-G687 ◽

Cited By ~ 34

Author(s):

Ann Marie Gallagher ◽

Roberta A. Gottlieb

Keyword(s):

Cell Proliferation ◽

Small Intestine ◽

Cell Migration ◽

Epithelial Cell ◽

Epithelial Cells ◽

Intestinal Epithelial ◽

Transmembrane Conductance Regulator ◽

Epithelial Cell Proliferation ◽

Transmembrane Conductance ◽

Epithelial Cell Migration

Expression of a mutated cystic fibrosis transmembrane conductance regulator (CFTR) has been shown to enhance proliferation within CF airways, and cells expressing a mutated CFTR have been shown to be less susceptible to apoptosis. Because the CFTR is expressed in the epithelial cells lining the gastrointestinal tract and all CF mouse models are characterized by gastrointestinal obstruction, we hypothesized that CFTR null mice would have increased epithelial cell proliferation and reduced apoptosis within the small intestine. The rate of intestinal epithelial cell migration from crypt to villus was increased in CFTR null mice relative to mice expressing the wild-type CFTR. This difference in migration could be explained by an increase in epithelial cell proliferation but not by a difference in apoptosis within the crypts of Lieberkühn. In addition, using two independent sets of CF cell lines, we found that epithelial cell susceptibility to apoptosis was unrelated to the presence of a functional CFTR. Thus increased proliferation but not alterations in apoptosis within epithelial cells might contribute to the pathophysiology of CF.

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Epidermal growth factor stimulates Rac activation through Src and phosphatidylinositol 3-kinase to promote colonic epithelial cell migration

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00340.2007 ◽

2008 ◽

Vol 294 (1) ◽

pp. G276-G285 ◽

Cited By ~ 66

Author(s):

Rebecca S. Dise ◽

Mark R. Frey ◽

Robert H. Whitehead ◽

D. Brent Polk

Keyword(s):

Wound Healing ◽

Cell Migration ◽

Epithelial Cell ◽

Intestinal Epithelial Cell ◽

Wound Closure ◽

Intestinal Epithelial ◽

Phosphatidylinositol 3 Kinase ◽

Epithelial Cell Migration ◽

Wound Margin ◽

Epidermal Growth

Regulated intestinal epithelial cell migration plays a key role in wound healing and maintenance of a healthy gastrointestinal tract. Epidermal growth factor (EGF) stimulates cell migration and wound closure in intestinal epithelial cells through incompletely understood mechanisms. In this study we investigated the role of the small GTPase Rac in EGF-induced cell migration using an in vitro wound-healing assay. In mouse colonic epithelial (MCE) cell lines, EGF-stimulated wound closure was accompanied by a doubling of the number of cells containing lamellipodial extensions at the wound margin, increased Rac membrane translocation in cells at the wound margin, and rapid Rac activation. Either Rac1 small interfering (si)RNA or a Rac1 inhibitor completely blocked EGF-stimulated wound closure. Whereas EGF failed to activate Rac in colon cells from EGF receptor (EGFR) knockout mice, stable expression of wild-type EGFR restored EGF-stimulated Rac activation and migration. Pharmacological inhibition of either phosphatidylinositol 3-kinase (PI3K) or Src family kinases reduced EGF-stimulated Rac activation. Cotreatment of cells with both inhibitors completely blocked EGF-stimulated Rac activation and localization to the leading edge of cells and lamellipodial extension. Our results present a novel mechanism by which the PI3K and Src signaling cascades cooperate to activate Rac and promote intestinal epithelial cell migration downstream of EGFR.

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