Limits...
MMP-9 and CXCL8/IL-8 are potential therapeutic targets in epidermolysis bullosa simplex.

Lettner T, Lang R, Klausegger A, Hainzl S, Bauer JW, Wally V - PLoS ONE (2013)

Bottom Line: We found kallikrein-related peptidases and matrix metalloproteinases to be upregulated.We also found elevated expression of chemokines, and we observed deregulation of the Cdc42 pathway as well as aberrant expression of cytokeratins and junction proteins.We further demonstrated, that expression of these genes is dependent on interleukin-1 β signaling.

View Article: PubMed Central - PubMed

Affiliation: Division of Experimental Dermatology and EB House Austria, Salzburg, Austria. thomas.lettner@gmail.com

ABSTRACT
Epidermolysis bullosa refers to a group of genodermatoses that affects the integrity of epithelial layers, phenotypically resulting in severe skin blistering. Dowling-Meara, the major subtype of epidermolysis bullosa simplex, is inherited in an autosomal dominant manner and can be caused by mutations in either the keratin-5 (K5) or the keratin-14 (K14) gene. Currently, no therapeutic approach is known, and the main objective of this study was to identify novel therapeutic targets. We used microarray analysis, semi-quantitative real-time PCR, western blot and ELISA to identify differentially regulated genes in two K14 mutant cell lines carrying the mutations K14 R125P and K14 R125H, respectively. We found kallikrein-related peptidases and matrix metalloproteinases to be upregulated. We also found elevated expression of chemokines, and we observed deregulation of the Cdc42 pathway as well as aberrant expression of cytokeratins and junction proteins. We further demonstrated, that expression of these genes is dependent on interleukin-1 β signaling. To evaluate these data in vivo we analysed the blister fluids of epidermolysis bullosa simplex patients vs. healthy controls and identified matrix metalloproteinase-9 and the chemokine CXCL8/IL-8 as potential therapeutic targets.

Show MeSH

Related in: MedlinePlus

Reduced expression of target genes upon IL-1β depletion.NEB-1 cells and EBDM-1 cells were incubated with 2 µg/ml IL-1β neutralizing antibody for 24 h to deplete IL-1β in the culture medium. Expression of untreated EBDM-1 cells (−) was compared to treated EBDM-1 cells (+) by SQRT-PCR. We picked at least one representative target gene of each of the six functional groups identified in the microarray. All investigated targets showed significant downregulation after IL-1β depletion. No differences in geneexpression were observed in NEB-1 cells −/+ antibody incubation (n = 3). Student’s t-test was performed with p values: * ≤0.05, *** ≤0.005, ΔΔ ≤0.0005.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3716611&req=5

pone-0070123-g007: Reduced expression of target genes upon IL-1β depletion.NEB-1 cells and EBDM-1 cells were incubated with 2 µg/ml IL-1β neutralizing antibody for 24 h to deplete IL-1β in the culture medium. Expression of untreated EBDM-1 cells (−) was compared to treated EBDM-1 cells (+) by SQRT-PCR. We picked at least one representative target gene of each of the six functional groups identified in the microarray. All investigated targets showed significant downregulation after IL-1β depletion. No differences in geneexpression were observed in NEB-1 cells −/+ antibody incubation (n = 3). Student’s t-test was performed with p values: * ≤0.05, *** ≤0.005, ΔΔ ≤0.0005.

Mentions: To study the role of IL-1β and its ability to alter the gene expression profile, we incubated EBDM-1 cells with IL-1β neutralizing antibody (2 µg/ml, R&D Systems) for 24 h and analyzed mRNA expression of distinct target genes by using SQRT-PCR. We chose EBDM-1 because it showed the most severe phenotype in most of our experiments. As targets we chose at least one representative of each of the six groups of regulated genes identified in our microarray analysis. In three independent experiments and with at least four SQRT-PCR runs per experiment, we observed a significant reduction of gene expression after IL-1β antibody incubation for all of the investigated target genes (Fig. 7).


MMP-9 and CXCL8/IL-8 are potential therapeutic targets in epidermolysis bullosa simplex.

Lettner T, Lang R, Klausegger A, Hainzl S, Bauer JW, Wally V - PLoS ONE (2013)

Reduced expression of target genes upon IL-1β depletion.NEB-1 cells and EBDM-1 cells were incubated with 2 µg/ml IL-1β neutralizing antibody for 24 h to deplete IL-1β in the culture medium. Expression of untreated EBDM-1 cells (−) was compared to treated EBDM-1 cells (+) by SQRT-PCR. We picked at least one representative target gene of each of the six functional groups identified in the microarray. All investigated targets showed significant downregulation after IL-1β depletion. No differences in geneexpression were observed in NEB-1 cells −/+ antibody incubation (n = 3). Student’s t-test was performed with p values: * ≤0.05, *** ≤0.005, ΔΔ ≤0.0005.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3716611&req=5

pone-0070123-g007: Reduced expression of target genes upon IL-1β depletion.NEB-1 cells and EBDM-1 cells were incubated with 2 µg/ml IL-1β neutralizing antibody for 24 h to deplete IL-1β in the culture medium. Expression of untreated EBDM-1 cells (−) was compared to treated EBDM-1 cells (+) by SQRT-PCR. We picked at least one representative target gene of each of the six functional groups identified in the microarray. All investigated targets showed significant downregulation after IL-1β depletion. No differences in geneexpression were observed in NEB-1 cells −/+ antibody incubation (n = 3). Student’s t-test was performed with p values: * ≤0.05, *** ≤0.005, ΔΔ ≤0.0005.
Mentions: To study the role of IL-1β and its ability to alter the gene expression profile, we incubated EBDM-1 cells with IL-1β neutralizing antibody (2 µg/ml, R&D Systems) for 24 h and analyzed mRNA expression of distinct target genes by using SQRT-PCR. We chose EBDM-1 because it showed the most severe phenotype in most of our experiments. As targets we chose at least one representative of each of the six groups of regulated genes identified in our microarray analysis. In three independent experiments and with at least four SQRT-PCR runs per experiment, we observed a significant reduction of gene expression after IL-1β antibody incubation for all of the investigated target genes (Fig. 7).

Bottom Line: We found kallikrein-related peptidases and matrix metalloproteinases to be upregulated.We also found elevated expression of chemokines, and we observed deregulation of the Cdc42 pathway as well as aberrant expression of cytokeratins and junction proteins.We further demonstrated, that expression of these genes is dependent on interleukin-1 β signaling.

View Article: PubMed Central - PubMed

Affiliation: Division of Experimental Dermatology and EB House Austria, Salzburg, Austria. thomas.lettner@gmail.com

ABSTRACT
Epidermolysis bullosa refers to a group of genodermatoses that affects the integrity of epithelial layers, phenotypically resulting in severe skin blistering. Dowling-Meara, the major subtype of epidermolysis bullosa simplex, is inherited in an autosomal dominant manner and can be caused by mutations in either the keratin-5 (K5) or the keratin-14 (K14) gene. Currently, no therapeutic approach is known, and the main objective of this study was to identify novel therapeutic targets. We used microarray analysis, semi-quantitative real-time PCR, western blot and ELISA to identify differentially regulated genes in two K14 mutant cell lines carrying the mutations K14 R125P and K14 R125H, respectively. We found kallikrein-related peptidases and matrix metalloproteinases to be upregulated. We also found elevated expression of chemokines, and we observed deregulation of the Cdc42 pathway as well as aberrant expression of cytokeratins and junction proteins. We further demonstrated, that expression of these genes is dependent on interleukin-1 β signaling. To evaluate these data in vivo we analysed the blister fluids of epidermolysis bullosa simplex patients vs. healthy controls and identified matrix metalloproteinase-9 and the chemokine CXCL8/IL-8 as potential therapeutic targets.

Show MeSH
Related in: MedlinePlus