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Ectodomain shedding of epidermal growth factor receptor ligands is required for keratinocyte migration in cutaneous wound healing.

Tokumaru S, Higashiyama S, Endo T, Nakagawa T, Miyagawa JI, Yamamori K, Hanakawa Y, Ohmoto H, Yoshino K, Shirakata Y, Matsuzawa Y, Hashimoto K, Taniguchi N - J. Cell Biol. (2000)

Bottom Line: Keratinocyte proliferation and migration are essential to cutaneous wound healing and are, in part, mediated in an autocrine fashion by epidermal growth factor receptor (EGFR)-ligand interactions.Soluble EGFR-immunoglobulin G-Fcgamma fusion protein, which is able to neutralize all EGFR ligands, also suppressed keratinocyte migration in vitro.The application of OSU8-1 to wound sites in mice greatly retarded reepithelialization as the result of a failure in keratinocyte migration, but this effect could be overcome if recombinant soluble HB-EGF was added along with OSU8-1.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Osaka University Medical School, Suita, Osaka 565-0871, Japan.

ABSTRACT
Keratinocyte proliferation and migration are essential to cutaneous wound healing and are, in part, mediated in an autocrine fashion by epidermal growth factor receptor (EGFR)-ligand interactions. EGFR ligands are initially synthesized as membrane-anchored forms, but can be processed and shed as soluble forms. We provide evidence here that wound stimuli induce keratinocyte shedding of EGFR ligands in vitro, particularly the ligand heparin-binding EGF-like growth factor (HB-EGF). The resulting soluble ligands stimulated transient activation of EGFR. OSU8-1, an inhibitor of EGFR ligand shedding, abrogated the wound-induced activation of EGFR and caused suppression of keratinocyte migration in vitro. Soluble EGFR-immunoglobulin G-Fcgamma fusion protein, which is able to neutralize all EGFR ligands, also suppressed keratinocyte migration in vitro. The application of OSU8-1 to wound sites in mice greatly retarded reepithelialization as the result of a failure in keratinocyte migration, but this effect could be overcome if recombinant soluble HB-EGF was added along with OSU8-1. These findings indicate that the shedding of EGFR ligands represents a critical event in keratinocyte migration, and suggest their possible use as an effective clinical treatment in the early phases of wound healing.

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Effect of OSU8-1 on wound healing in vivo. Histology of excised mouse skin on day 6 after wounding, without or with 10 mM OSU8-1 treatment. Keratinocytes were stained with antikeratin/cytokeratin antibody. (a) A typical tissue section from the control wounds (n = 15). Keratinocytes are observed to have migrated from the edge of the wound, spreading under the crust and covering the wound site. Arrowheads indicate the base of the keratinocyte layer. The arrow marks the edge of the wound. (b) A typical tissue section from an OSU8-1–treated wound (n = 10). Both keratinocyte migration and the regeneration of the epidermis were greatly inhibited. (c) A typical tissue section from a wound treated with a cocktail of OSU8-1 and HB-EGF (5 μg/ml; n = 5). Both keratinocyte migration and the regeneration of the epidermis were completely restored.
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Figure 8: Effect of OSU8-1 on wound healing in vivo. Histology of excised mouse skin on day 6 after wounding, without or with 10 mM OSU8-1 treatment. Keratinocytes were stained with antikeratin/cytokeratin antibody. (a) A typical tissue section from the control wounds (n = 15). Keratinocytes are observed to have migrated from the edge of the wound, spreading under the crust and covering the wound site. Arrowheads indicate the base of the keratinocyte layer. The arrow marks the edge of the wound. (b) A typical tissue section from an OSU8-1–treated wound (n = 10). Both keratinocyte migration and the regeneration of the epidermis were greatly inhibited. (c) A typical tissue section from a wound treated with a cocktail of OSU8-1 and HB-EGF (5 μg/ml; n = 5). Both keratinocyte migration and the regeneration of the epidermis were completely restored.

Mentions: Under conditions of wounding, keratinocytes migrate from hair follicles and the wound edge towards the center of the wound. Given the effects of OSU8-1 described above on keratinocyte migration in vitro in response to wound stimuli, we therefore investigated the effects of OSU8-1 on cutaneous wound healing in mice. Keratin staining of wound tissue sections from wounds treated with vehicle solution alone revealed the migration of keratinocytes into the wounded site. Regenerated epithelium and granulation tissue was observed in the upper dermis at day 6 after wounding (n = 15, Fig. 8 a). In contrast, wound tissue sections from wounds treated with 10 mM of OSU8-1 showed essentially no tissue regeneration or repair of the epidermis (Fig. 8 b). It is noteworthy that the migration of keratinocytes into the wounded site was completely blocked in all of the OSU8-1– treated wounds (n = 10; Fig. 8 b). The application of HB-EGF at 5 μg/ml in addition to OSU8-1 restored keratinocyte migration, resulting in healing wounded sites (n = 5; Fig. 8 c). These results indicate that the production of the soluble forms of EGFR ligands represents a critical event in the process of wound healing in vivo.


Ectodomain shedding of epidermal growth factor receptor ligands is required for keratinocyte migration in cutaneous wound healing.

Tokumaru S, Higashiyama S, Endo T, Nakagawa T, Miyagawa JI, Yamamori K, Hanakawa Y, Ohmoto H, Yoshino K, Shirakata Y, Matsuzawa Y, Hashimoto K, Taniguchi N - J. Cell Biol. (2000)

Effect of OSU8-1 on wound healing in vivo. Histology of excised mouse skin on day 6 after wounding, without or with 10 mM OSU8-1 treatment. Keratinocytes were stained with antikeratin/cytokeratin antibody. (a) A typical tissue section from the control wounds (n = 15). Keratinocytes are observed to have migrated from the edge of the wound, spreading under the crust and covering the wound site. Arrowheads indicate the base of the keratinocyte layer. The arrow marks the edge of the wound. (b) A typical tissue section from an OSU8-1–treated wound (n = 10). Both keratinocyte migration and the regeneration of the epidermis were greatly inhibited. (c) A typical tissue section from a wound treated with a cocktail of OSU8-1 and HB-EGF (5 μg/ml; n = 5). Both keratinocyte migration and the regeneration of the epidermis were completely restored.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2192647&req=5

Figure 8: Effect of OSU8-1 on wound healing in vivo. Histology of excised mouse skin on day 6 after wounding, without or with 10 mM OSU8-1 treatment. Keratinocytes were stained with antikeratin/cytokeratin antibody. (a) A typical tissue section from the control wounds (n = 15). Keratinocytes are observed to have migrated from the edge of the wound, spreading under the crust and covering the wound site. Arrowheads indicate the base of the keratinocyte layer. The arrow marks the edge of the wound. (b) A typical tissue section from an OSU8-1–treated wound (n = 10). Both keratinocyte migration and the regeneration of the epidermis were greatly inhibited. (c) A typical tissue section from a wound treated with a cocktail of OSU8-1 and HB-EGF (5 μg/ml; n = 5). Both keratinocyte migration and the regeneration of the epidermis were completely restored.
Mentions: Under conditions of wounding, keratinocytes migrate from hair follicles and the wound edge towards the center of the wound. Given the effects of OSU8-1 described above on keratinocyte migration in vitro in response to wound stimuli, we therefore investigated the effects of OSU8-1 on cutaneous wound healing in mice. Keratin staining of wound tissue sections from wounds treated with vehicle solution alone revealed the migration of keratinocytes into the wounded site. Regenerated epithelium and granulation tissue was observed in the upper dermis at day 6 after wounding (n = 15, Fig. 8 a). In contrast, wound tissue sections from wounds treated with 10 mM of OSU8-1 showed essentially no tissue regeneration or repair of the epidermis (Fig. 8 b). It is noteworthy that the migration of keratinocytes into the wounded site was completely blocked in all of the OSU8-1– treated wounds (n = 10; Fig. 8 b). The application of HB-EGF at 5 μg/ml in addition to OSU8-1 restored keratinocyte migration, resulting in healing wounded sites (n = 5; Fig. 8 c). These results indicate that the production of the soluble forms of EGFR ligands represents a critical event in the process of wound healing in vivo.

Bottom Line: Keratinocyte proliferation and migration are essential to cutaneous wound healing and are, in part, mediated in an autocrine fashion by epidermal growth factor receptor (EGFR)-ligand interactions.Soluble EGFR-immunoglobulin G-Fcgamma fusion protein, which is able to neutralize all EGFR ligands, also suppressed keratinocyte migration in vitro.The application of OSU8-1 to wound sites in mice greatly retarded reepithelialization as the result of a failure in keratinocyte migration, but this effect could be overcome if recombinant soluble HB-EGF was added along with OSU8-1.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Osaka University Medical School, Suita, Osaka 565-0871, Japan.

ABSTRACT
Keratinocyte proliferation and migration are essential to cutaneous wound healing and are, in part, mediated in an autocrine fashion by epidermal growth factor receptor (EGFR)-ligand interactions. EGFR ligands are initially synthesized as membrane-anchored forms, but can be processed and shed as soluble forms. We provide evidence here that wound stimuli induce keratinocyte shedding of EGFR ligands in vitro, particularly the ligand heparin-binding EGF-like growth factor (HB-EGF). The resulting soluble ligands stimulated transient activation of EGFR. OSU8-1, an inhibitor of EGFR ligand shedding, abrogated the wound-induced activation of EGFR and caused suppression of keratinocyte migration in vitro. Soluble EGFR-immunoglobulin G-Fcgamma fusion protein, which is able to neutralize all EGFR ligands, also suppressed keratinocyte migration in vitro. The application of OSU8-1 to wound sites in mice greatly retarded reepithelialization as the result of a failure in keratinocyte migration, but this effect could be overcome if recombinant soluble HB-EGF was added along with OSU8-1. These findings indicate that the shedding of EGFR ligands represents a critical event in keratinocyte migration, and suggest their possible use as an effective clinical treatment in the early phases of wound healing.

Show MeSH
Related in: MedlinePlus