Development of Allele-Specific Therapeutic siRNA for Keratin 5 Mutations in Epidermolysis Bullosa Simplex.

Sarah Atkinson, Victoria McGilligan, H Liao, I Szeverenyi, FJ Smith, Tara Moore, WH McLean

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Epidermolysis bullosa simplex (EBS) is an incurable, inherited skin-blistering disorder predominantly caused by dominant-negative mutations in the genes encoding keratins K5 or K14. RNA interference, particularly in the form of small interfering RNA (siRNA), offers a potential therapy route for EBS and related keratin disorders by selectively silencing the mutant allele. Here, using a systemic screening system based on a luciferase reporter gene assay, we have developed mutant-specific siRNAs for two independent EBS-causing missense mutations in the K5 gene (p.Ser181Pro and p.Asn193Lys). The specificity of the allele-specific inhibitors identified in the screen was subsequently confirmed at the protein level, where the lead inhibitors were shown to strongly knock down the expression of mutant proteins with negligible effect on wild-type K5 expression. In a cell-based model system, the lead inhibitors were able to significantly reverse the cytoskeletal aggregation phenotype. Overall, this approach shows promise for the treatment of EBS and paves the way for future clinical trials.
LanguageEnglish
Pages2079-2086
JournalJournal of Investigative Dermatology
Volume131
Issue number10
Early online date30 Jun 2011
DOIs
Publication statusPublished - Oct 2011

Fingerprint

Epidermolysis Bullosa Simplex
Keratin-5
Keratins
Small Interfering RNA
Genes
Alleles
Mutation
Gene encoding
Mutant Proteins
Luciferases
Assays
Skin
Screening
Agglomeration
RNA
Missense Mutation
Therapeutics
RNA Interference
Reporter Genes
Proteins

Cite this

@article{fd4cdcbf2fd840c38f665094dd57dcf2,
title = "Development of Allele-Specific Therapeutic siRNA for Keratin 5 Mutations in Epidermolysis Bullosa Simplex.",
abstract = "Epidermolysis bullosa simplex (EBS) is an incurable, inherited skin-blistering disorder predominantly caused by dominant-negative mutations in the genes encoding keratins K5 or K14. RNA interference, particularly in the form of small interfering RNA (siRNA), offers a potential therapy route for EBS and related keratin disorders by selectively silencing the mutant allele. Here, using a systemic screening system based on a luciferase reporter gene assay, we have developed mutant-specific siRNAs for two independent EBS-causing missense mutations in the K5 gene (p.Ser181Pro and p.Asn193Lys). The specificity of the allele-specific inhibitors identified in the screen was subsequently confirmed at the protein level, where the lead inhibitors were shown to strongly knock down the expression of mutant proteins with negligible effect on wild-type K5 expression. In a cell-based model system, the lead inhibitors were able to significantly reverse the cytoskeletal aggregation phenotype. Overall, this approach shows promise for the treatment of EBS and paves the way for future clinical trials.",
author = "Sarah Atkinson and Victoria McGilligan and H Liao and I Szeverenyi and FJ Smith and Tara Moore and WH McLean",
year = "2011",
month = "10",
doi = "10.1038/jid.2011.169",
language = "English",
volume = "131",
pages = "2079--2086",
journal = "Journal of Investigative Dermatology",
issn = "0022-202X",
publisher = "Elsevier",
number = "10",

}

Development of Allele-Specific Therapeutic siRNA for Keratin 5 Mutations in Epidermolysis Bullosa Simplex. / Atkinson, Sarah; McGilligan, Victoria; Liao, H; Szeverenyi, I; Smith, FJ; Moore, Tara; McLean, WH.

In: Journal of Investigative Dermatology, Vol. 131, No. 10, 10.2011, p. 2079-2086.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Development of Allele-Specific Therapeutic siRNA for Keratin 5 Mutations in Epidermolysis Bullosa Simplex.

AU - Atkinson, Sarah

AU - McGilligan, Victoria

AU - Liao, H

AU - Szeverenyi, I

AU - Smith, FJ

AU - Moore, Tara

AU - McLean, WH

PY - 2011/10

Y1 - 2011/10

N2 - Epidermolysis bullosa simplex (EBS) is an incurable, inherited skin-blistering disorder predominantly caused by dominant-negative mutations in the genes encoding keratins K5 or K14. RNA interference, particularly in the form of small interfering RNA (siRNA), offers a potential therapy route for EBS and related keratin disorders by selectively silencing the mutant allele. Here, using a systemic screening system based on a luciferase reporter gene assay, we have developed mutant-specific siRNAs for two independent EBS-causing missense mutations in the K5 gene (p.Ser181Pro and p.Asn193Lys). The specificity of the allele-specific inhibitors identified in the screen was subsequently confirmed at the protein level, where the lead inhibitors were shown to strongly knock down the expression of mutant proteins with negligible effect on wild-type K5 expression. In a cell-based model system, the lead inhibitors were able to significantly reverse the cytoskeletal aggregation phenotype. Overall, this approach shows promise for the treatment of EBS and paves the way for future clinical trials.

AB - Epidermolysis bullosa simplex (EBS) is an incurable, inherited skin-blistering disorder predominantly caused by dominant-negative mutations in the genes encoding keratins K5 or K14. RNA interference, particularly in the form of small interfering RNA (siRNA), offers a potential therapy route for EBS and related keratin disorders by selectively silencing the mutant allele. Here, using a systemic screening system based on a luciferase reporter gene assay, we have developed mutant-specific siRNAs for two independent EBS-causing missense mutations in the K5 gene (p.Ser181Pro and p.Asn193Lys). The specificity of the allele-specific inhibitors identified in the screen was subsequently confirmed at the protein level, where the lead inhibitors were shown to strongly knock down the expression of mutant proteins with negligible effect on wild-type K5 expression. In a cell-based model system, the lead inhibitors were able to significantly reverse the cytoskeletal aggregation phenotype. Overall, this approach shows promise for the treatment of EBS and paves the way for future clinical trials.

U2 - 10.1038/jid.2011.169

DO - 10.1038/jid.2011.169

M3 - Article

VL - 131

SP - 2079

EP - 2086

JO - Journal of Investigative Dermatology

T2 - Journal of Investigative Dermatology

JF - Journal of Investigative Dermatology

SN - 0022-202X

IS - 10

ER -