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PAR2 absence completely rescues inflammation and ichthyosis caused by altered CAP1/Prss8 expression in mouse skin.

Frateschi S, Camerer E, Crisante G, Rieser S, Membrez M, Charles RP, Beermann F, Stehle JC, Breiden B, Sandhoff K, Rotman S, Haftek M, Wilson A, Ryser S, Steinhoff M, Coughlin SR, Hummler E - Nat Commun (2011)

Bottom Line: K14-CAP1/Prss8 ectopic expression impairs epidermal barrier function and causes skin inflammation characterized by an increase in thymic stromal lymphopoietin levels and immune cell infiltrations.Strikingly, both gross and functional K14-CAP1/Prss8-induced phenotypes are completely negated when superimposed on a PAR2- background, establishing PAR2 as a pivotal mediator of pathogenesis.Our data provide genetic evidence for PAR2 as a downstream effector of CAP1/Prss8 in a signalling cascade that may provide novel therapeutic targets for ichthyoses, pruritus and inflammatory skin diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Toxicology, University of Lausanne, Lausanne 1005, Switzerland.

ABSTRACT
Altered serine protease activity is associated with skin disorders in humans and in mice. The serine protease channel-activating protease-1 (CAP1; also termed protease serine S1 family member 8 (Prss8)) is important for epidermal homeostasis and is thus indispensable for postnatal survival in mice, but its roles and effectors in skin pathology are poorly defined. In this paper, we report that transgenic expression in mouse skin of either CAP1/Prss8 (K14-CAP1/Prss8) or protease-activated receptor-2 (PAR2; Grhl3(PAR2/+)), one candidate downstream target, causes epidermal hyperplasia, ichthyosis and itching. K14-CAP1/Prss8 ectopic expression impairs epidermal barrier function and causes skin inflammation characterized by an increase in thymic stromal lymphopoietin levels and immune cell infiltrations. Strikingly, both gross and functional K14-CAP1/Prss8-induced phenotypes are completely negated when superimposed on a PAR2- background, establishing PAR2 as a pivotal mediator of pathogenesis. Our data provide genetic evidence for PAR2 as a downstream effector of CAP1/Prss8 in a signalling cascade that may provide novel therapeutic targets for ichthyoses, pruritus and inflammatory skin diseases.

No MeSH data available.


Related in: MedlinePlus

K14-CAP1/Prss8 mice exhibit itching and severe skin inflammation.(a) Skin injuries with open wounds and lymph node inflammation (haematoxylin and eosin staining, bar represents 20 μm) in a representative 7-month-old transgenic mouse. (b) Dot plot indicating the duration of the scratching behaviour expressed in percentage. Each dot represents a single event per animal. Bars indicate the average of the time spent scratching, P<0.01. (c) Semiquantitative RT–PCR analysis of TSLP, MMP9, IL-1β and IL-1α (30 cycles) in transgenic versus wild-type skin. The reaction is controlled by detection of β-actin (25 cycles). Water was used as negative control. Data are representative of n=3 mice per genotype. (d) Giemsa, periodic acid-Schiff staining and S100, F4/80 and CD3 labelling in skin of 2-week-old wild-type and K14-CAP1/Prss8 transgenic mice. Pictures are representative of n=3 animals per genotype. Bars represent 20 μm. (e) Quantification of the number of CD3-positive cells in transgenic versus wild-type skin. n=3 mice per genotype, *P<0.01. (f) FACS analysis of dendritic cells (CD11c+), macrophages (F4/80+) and Tγ cells (CD3+; CD3+/Vg3+; and CD3+/Vg3-) in skin of 6-day-old pups (n=3 mice per genotype). (g) F4/80 and S100-positive cells in skin of 2-day-old transgenic and littermate controls. Bar represents 20 μm; pictures are representative of n=3 mice per genotype. (h) Quantification of the number of S100-positive cells in 2-day-old transgenic and littermate animals. n=3 mice per genotype, *P<0.05. (i) TSLP quantitative RT–PCR analyses of wild-type and transgenic skin in 2-day-old pups (n≥3 mice per group, *P<0.05, **P<0.01). (j) TEWL and body weight (k) measurements at P1, P2 and P5 (n≥4 mice per group). (l) Quantitative RT–PCR examination of TSLP, IL1α, IL1β and MMP9 in transgenic primary keratinocytes, ***P<0.001. (m) Quantification of cell growth assessed in primary keratinocytes derived from 2-day-old transgenic and control littermates. (l, m) The data are representative of three independent experiments. All data are presented as mean±s.e.m.
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f4: K14-CAP1/Prss8 mice exhibit itching and severe skin inflammation.(a) Skin injuries with open wounds and lymph node inflammation (haematoxylin and eosin staining, bar represents 20 μm) in a representative 7-month-old transgenic mouse. (b) Dot plot indicating the duration of the scratching behaviour expressed in percentage. Each dot represents a single event per animal. Bars indicate the average of the time spent scratching, P<0.01. (c) Semiquantitative RT–PCR analysis of TSLP, MMP9, IL-1β and IL-1α (30 cycles) in transgenic versus wild-type skin. The reaction is controlled by detection of β-actin (25 cycles). Water was used as negative control. Data are representative of n=3 mice per genotype. (d) Giemsa, periodic acid-Schiff staining and S100, F4/80 and CD3 labelling in skin of 2-week-old wild-type and K14-CAP1/Prss8 transgenic mice. Pictures are representative of n=3 animals per genotype. Bars represent 20 μm. (e) Quantification of the number of CD3-positive cells in transgenic versus wild-type skin. n=3 mice per genotype, *P<0.01. (f) FACS analysis of dendritic cells (CD11c+), macrophages (F4/80+) and Tγ cells (CD3+; CD3+/Vg3+; and CD3+/Vg3-) in skin of 6-day-old pups (n=3 mice per genotype). (g) F4/80 and S100-positive cells in skin of 2-day-old transgenic and littermate controls. Bar represents 20 μm; pictures are representative of n=3 mice per genotype. (h) Quantification of the number of S100-positive cells in 2-day-old transgenic and littermate animals. n=3 mice per genotype, *P<0.05. (i) TSLP quantitative RT–PCR analyses of wild-type and transgenic skin in 2-day-old pups (n≥3 mice per group, *P<0.05, **P<0.01). (j) TEWL and body weight (k) measurements at P1, P2 and P5 (n≥4 mice per group). (l) Quantitative RT–PCR examination of TSLP, IL1α, IL1β and MMP9 in transgenic primary keratinocytes, ***P<0.001. (m) Quantification of cell growth assessed in primary keratinocytes derived from 2-day-old transgenic and control littermates. (l, m) The data are representative of three independent experiments. All data are presented as mean±s.e.m.

Mentions: Transgenic animals surviving till adulthood frequently exhibited skin lesions (Fig. 4a; Supplementary Fig. S1m) possibly caused or aggravated by increased scratching behaviour (Fig. 4b; Supplementary Fig. S1n), which was evident after weaning and persistent throughout life. Lesions were sometimes accompanied by inflammation and swelling of regional lymph nodes, which were found to be enriched in plasma cells (Fig. 4a; Supplementary Fig. S1m).


PAR2 absence completely rescues inflammation and ichthyosis caused by altered CAP1/Prss8 expression in mouse skin.

Frateschi S, Camerer E, Crisante G, Rieser S, Membrez M, Charles RP, Beermann F, Stehle JC, Breiden B, Sandhoff K, Rotman S, Haftek M, Wilson A, Ryser S, Steinhoff M, Coughlin SR, Hummler E - Nat Commun (2011)

K14-CAP1/Prss8 mice exhibit itching and severe skin inflammation.(a) Skin injuries with open wounds and lymph node inflammation (haematoxylin and eosin staining, bar represents 20 μm) in a representative 7-month-old transgenic mouse. (b) Dot plot indicating the duration of the scratching behaviour expressed in percentage. Each dot represents a single event per animal. Bars indicate the average of the time spent scratching, P<0.01. (c) Semiquantitative RT–PCR analysis of TSLP, MMP9, IL-1β and IL-1α (30 cycles) in transgenic versus wild-type skin. The reaction is controlled by detection of β-actin (25 cycles). Water was used as negative control. Data are representative of n=3 mice per genotype. (d) Giemsa, periodic acid-Schiff staining and S100, F4/80 and CD3 labelling in skin of 2-week-old wild-type and K14-CAP1/Prss8 transgenic mice. Pictures are representative of n=3 animals per genotype. Bars represent 20 μm. (e) Quantification of the number of CD3-positive cells in transgenic versus wild-type skin. n=3 mice per genotype, *P<0.01. (f) FACS analysis of dendritic cells (CD11c+), macrophages (F4/80+) and Tγ cells (CD3+; CD3+/Vg3+; and CD3+/Vg3-) in skin of 6-day-old pups (n=3 mice per genotype). (g) F4/80 and S100-positive cells in skin of 2-day-old transgenic and littermate controls. Bar represents 20 μm; pictures are representative of n=3 mice per genotype. (h) Quantification of the number of S100-positive cells in 2-day-old transgenic and littermate animals. n=3 mice per genotype, *P<0.05. (i) TSLP quantitative RT–PCR analyses of wild-type and transgenic skin in 2-day-old pups (n≥3 mice per group, *P<0.05, **P<0.01). (j) TEWL and body weight (k) measurements at P1, P2 and P5 (n≥4 mice per group). (l) Quantitative RT–PCR examination of TSLP, IL1α, IL1β and MMP9 in transgenic primary keratinocytes, ***P<0.001. (m) Quantification of cell growth assessed in primary keratinocytes derived from 2-day-old transgenic and control littermates. (l, m) The data are representative of three independent experiments. All data are presented as mean±s.e.m.
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f4: K14-CAP1/Prss8 mice exhibit itching and severe skin inflammation.(a) Skin injuries with open wounds and lymph node inflammation (haematoxylin and eosin staining, bar represents 20 μm) in a representative 7-month-old transgenic mouse. (b) Dot plot indicating the duration of the scratching behaviour expressed in percentage. Each dot represents a single event per animal. Bars indicate the average of the time spent scratching, P<0.01. (c) Semiquantitative RT–PCR analysis of TSLP, MMP9, IL-1β and IL-1α (30 cycles) in transgenic versus wild-type skin. The reaction is controlled by detection of β-actin (25 cycles). Water was used as negative control. Data are representative of n=3 mice per genotype. (d) Giemsa, periodic acid-Schiff staining and S100, F4/80 and CD3 labelling in skin of 2-week-old wild-type and K14-CAP1/Prss8 transgenic mice. Pictures are representative of n=3 animals per genotype. Bars represent 20 μm. (e) Quantification of the number of CD3-positive cells in transgenic versus wild-type skin. n=3 mice per genotype, *P<0.01. (f) FACS analysis of dendritic cells (CD11c+), macrophages (F4/80+) and Tγ cells (CD3+; CD3+/Vg3+; and CD3+/Vg3-) in skin of 6-day-old pups (n=3 mice per genotype). (g) F4/80 and S100-positive cells in skin of 2-day-old transgenic and littermate controls. Bar represents 20 μm; pictures are representative of n=3 mice per genotype. (h) Quantification of the number of S100-positive cells in 2-day-old transgenic and littermate animals. n=3 mice per genotype, *P<0.05. (i) TSLP quantitative RT–PCR analyses of wild-type and transgenic skin in 2-day-old pups (n≥3 mice per group, *P<0.05, **P<0.01). (j) TEWL and body weight (k) measurements at P1, P2 and P5 (n≥4 mice per group). (l) Quantitative RT–PCR examination of TSLP, IL1α, IL1β and MMP9 in transgenic primary keratinocytes, ***P<0.001. (m) Quantification of cell growth assessed in primary keratinocytes derived from 2-day-old transgenic and control littermates. (l, m) The data are representative of three independent experiments. All data are presented as mean±s.e.m.
Mentions: Transgenic animals surviving till adulthood frequently exhibited skin lesions (Fig. 4a; Supplementary Fig. S1m) possibly caused or aggravated by increased scratching behaviour (Fig. 4b; Supplementary Fig. S1n), which was evident after weaning and persistent throughout life. Lesions were sometimes accompanied by inflammation and swelling of regional lymph nodes, which were found to be enriched in plasma cells (Fig. 4a; Supplementary Fig. S1m).

Bottom Line: K14-CAP1/Prss8 ectopic expression impairs epidermal barrier function and causes skin inflammation characterized by an increase in thymic stromal lymphopoietin levels and immune cell infiltrations.Strikingly, both gross and functional K14-CAP1/Prss8-induced phenotypes are completely negated when superimposed on a PAR2- background, establishing PAR2 as a pivotal mediator of pathogenesis.Our data provide genetic evidence for PAR2 as a downstream effector of CAP1/Prss8 in a signalling cascade that may provide novel therapeutic targets for ichthyoses, pruritus and inflammatory skin diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Toxicology, University of Lausanne, Lausanne 1005, Switzerland.

ABSTRACT
Altered serine protease activity is associated with skin disorders in humans and in mice. The serine protease channel-activating protease-1 (CAP1; also termed protease serine S1 family member 8 (Prss8)) is important for epidermal homeostasis and is thus indispensable for postnatal survival in mice, but its roles and effectors in skin pathology are poorly defined. In this paper, we report that transgenic expression in mouse skin of either CAP1/Prss8 (K14-CAP1/Prss8) or protease-activated receptor-2 (PAR2; Grhl3(PAR2/+)), one candidate downstream target, causes epidermal hyperplasia, ichthyosis and itching. K14-CAP1/Prss8 ectopic expression impairs epidermal barrier function and causes skin inflammation characterized by an increase in thymic stromal lymphopoietin levels and immune cell infiltrations. Strikingly, both gross and functional K14-CAP1/Prss8-induced phenotypes are completely negated when superimposed on a PAR2- background, establishing PAR2 as a pivotal mediator of pathogenesis. Our data provide genetic evidence for PAR2 as a downstream effector of CAP1/Prss8 in a signalling cascade that may provide novel therapeutic targets for ichthyoses, pruritus and inflammatory skin diseases.

No MeSH data available.


Related in: MedlinePlus