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In Vitro Models for the Evaluation of Antisense Oligonucleotides in Skin
Jeroen Bremer1,2, Peter C van den Akker3,4,5
1University of Groningen, University Medical Center Groningen, Department of Genetics, Groningen, The Netherlands. j.bremer@umcg.nl.
Abstract:
The genodermatosis dystrophic epidermolysis bullosa (DEB) is caused by mutations in the COL7A1 gene which encodes type VII collagen (C7). In the cutaneous basement membrane zone, C7 secures attachment of the epidermal basal keratinocyte to the papillary dermis by means of anchoring fibril formation. The complete absence of these anchoring fibrils leads to severe blistering of skin and mucosa upon the slightest friction and early mortality. To date, although preclinical advances toward therapy are promising, treatment for the disease is merely symptomatic. Therefore, research into novel therapeutics is warranted.Antisense oligonucleotide (ASO)-mediated exon skipping is such a therapy . Clinical examination of naturally occurring exon skipping suggested that this mechanism could most likely benefit the most severely affected patients. The severe form of DEB is caused by biallelic null mutations. Exon skipping aims to bind an ASO to the mutated exon of the pre-mRNA in the cell nucleus. Thereby, the ASO inhibits the recognition of the mutated exon by the splicing machinery, and as a result, the mutated exon is spliced out from the mRNA with its surrounding introns, i.e., it is skipped. Here, we describe in vitro methods to evaluate ASO-mediated exon skipping in a preclinical setting.
Insights
Dystrophic epidermolysis bullosa (DEB) is a severe genetic skin disorder. This study explores antisense oligonucleotide-mediated exon skipping as a potential therapeutic strategy for DEB by evaluating in vitro methods.
Area of Science:
- Genetics
- Molecular Biology
- Dermatology
Background:
- Dystrophic epidermolysis bullosa (DEB) is a severe genodermatosis caused by COL7A1 gene mutations.
- Type VII collagen (C7) deficiency leads to defective anchoring fibrils, causing extreme skin fragility and early mortality.
- Current DEB treatments are symptomatic, necessitating novel therapeutic approaches.
Purpose of the Study:
- To establish in vitro methods for evaluating antisense oligonucleotide (ASO)-mediated exon skipping in a preclinical setting for DEB.
- To assess the potential of exon skipping as a therapeutic strategy for severe DEB cases.
Main Methods:
- Utilizing antisense oligonucleotides (ASOs) to target and skip mutated exons in COL7A1 pre-mRNA.
- Inhibiting the splicing machinery's recognition of mutated exons to promote their removal from mRNA.
- Developing in vitro assays to validate ASO efficacy in a preclinical context.
Main Results:
- The study focuses on describing the methodology for evaluating ASO-mediated exon skipping.
- The described methods are designed for preclinical assessment of this therapeutic approach.
Conclusions:
- Antisense oligonucleotide-mediated exon skipping shows promise as a therapeutic strategy for severe DEB.
- The developed in vitro methods are crucial for preclinical evaluation of this novel treatment approach.
