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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.

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Expression of junction proteins in EBS-DM cell lines.A. SQRT-PCR of desmocollin mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 4). DSC1 shows the highest increase in both EBS-DM cell lines. DSC2 and DSC3 are increased only in EBDM-1 but not in KEB-7. B. SQRT-PCR of desmoglein mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). DSG1 shows the highest increase in EBDM-1. Only a slight increase was observed for DSG3. DSG4 is only significantly increased in KEB-7. C. SQRT-PCR of gap junction protein mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). GJA1 and GJB6 expression is increased in both EBS-DM cell lines. GJB2 is only increased significantly in KEB-7. Student’s t-test was performed with p values: * ≤0.05, ** ≤0.01, *** ≤0.005, ‡ = no significant difference between investigated cell lines.
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pone-0070123-g005: Expression of junction proteins in EBS-DM cell lines.A. SQRT-PCR of desmocollin mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 4). DSC1 shows the highest increase in both EBS-DM cell lines. DSC2 and DSC3 are increased only in EBDM-1 but not in KEB-7. B. SQRT-PCR of desmoglein mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). DSG1 shows the highest increase in EBDM-1. Only a slight increase was observed for DSG3. DSG4 is only significantly increased in KEB-7. C. SQRT-PCR of gap junction protein mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). GJA1 and GJB6 expression is increased in both EBS-DM cell lines. GJB2 is only increased significantly in KEB-7. Student’s t-test was performed with p values: * ≤0.05, ** ≤0.01, *** ≤0.005, ‡ = no significant difference between investigated cell lines.

Mentions: Bioinformatic analysis of the microarray data revealed junction proteins as one major group of regulated genes (Table 6). We verified these data with SQRT-PCR for the desmocollins DSC1, DSC2 and DSC3 (Fig. 5A), the desmogleins DSG1, DSG3 and DSG4 (Fig. 5B), and the gap junction proteins GJA1, GJB2 and GJB6 (Fig. 5C) because they all showed significant upregulation in the array. The highest transcript levels were found for DSC1, DSG1 and GJB6 (connexin 30), with an increased expression of 15- to almost 30-fold compared to NEB-1 wild-type keratinocytes.


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)

Expression of junction proteins in EBS-DM cell lines.A. SQRT-PCR of desmocollin mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 4). DSC1 shows the highest increase in both EBS-DM cell lines. DSC2 and DSC3 are increased only in EBDM-1 but not in KEB-7. B. SQRT-PCR of desmoglein mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). DSG1 shows the highest increase in EBDM-1. Only a slight increase was observed for DSG3. DSG4 is only significantly increased in KEB-7. C. SQRT-PCR of gap junction protein mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). GJA1 and GJB6 expression is increased in both EBS-DM cell lines. GJB2 is only increased significantly in KEB-7. Student’s t-test was performed with p values: * ≤0.05, ** ≤0.01, *** ≤0.005, ‡ = no significant difference between investigated cell lines.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3716611&req=5

pone-0070123-g005: Expression of junction proteins in EBS-DM cell lines.A. SQRT-PCR of desmocollin mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 4). DSC1 shows the highest increase in both EBS-DM cell lines. DSC2 and DSC3 are increased only in EBDM-1 but not in KEB-7. B. SQRT-PCR of desmoglein mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). DSG1 shows the highest increase in EBDM-1. Only a slight increase was observed for DSG3. DSG4 is only significantly increased in KEB-7. C. SQRT-PCR of gap junction protein mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). GJA1 and GJB6 expression is increased in both EBS-DM cell lines. GJB2 is only increased significantly in KEB-7. Student’s t-test was performed with p values: * ≤0.05, ** ≤0.01, *** ≤0.005, ‡ = no significant difference between investigated cell lines.
Mentions: Bioinformatic analysis of the microarray data revealed junction proteins as one major group of regulated genes (Table 6). We verified these data with SQRT-PCR for the desmocollins DSC1, DSC2 and DSC3 (Fig. 5A), the desmogleins DSG1, DSG3 and DSG4 (Fig. 5B), and the gap junction proteins GJA1, GJB2 and GJB6 (Fig. 5C) because they all showed significant upregulation in the array. The highest transcript levels were found for DSC1, DSG1 and GJB6 (connexin 30), with an increased expression of 15- to almost 30-fold compared to NEB-1 wild-type keratinocytes.

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