Personalized Development of Antisense Oligonucleotides for Exon Skipping Restores Type XVII Collagen Expression in Junctional Epidermolysis Bullosa

• Published in: International journal of molecular sciences Vol. 22; no. 7; p. 3326
• Main Authors: Ablinger, Michael, Lettner, Thomas, Friedl, Nicole, Potocki, Hannah, Palmetzhofer, Theresa, Koller, Ulrich, Illmer, Julia, Liemberger, Bernadette, Hainzl, Stefan, Klausegger, Alfred, Reisenberger, Manuela, Lambert, Jo, Van Gele, Mireille, Desmet, Eline, Van Maelsaeke, Els, Wimmer, Monika, Zauner, Roland, Bauer, Johann W., Wally, Verena
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 24.03.2021; MDPI
• Subjects: Alternative Splicing; Autoantigens - metabolism; Binding sites; Biopsy; Cell Line; Cell Survival; Collagen; Collagen Type XVII; Epidermolysis Bullosa, Junctional - genetics; Epidermolysis Bullosa, Junctional - metabolism; Epidermolysis Bullosa, Junctional - therapy; Exons; Genes; Genotype; Homozygote; Humans; Keratinocytes - cytology; Liposomes - chemistry; Microscopy; Mutation; Non-Fibrillar Collagens - metabolism; Oligonucleotides, Antisense - genetics; Organ Culture Techniques; Patients; Proteins; RNA Splicing; RNA, Messenger - metabolism; Skin; junctional epidermolysis bullosa; splice mutation; molecular therapy; exon skipping; liposomes; antisense oligonucleotides; type XVII collagen; topical therapy
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms22073326

Intermediate junctional epidermolysis bullosa caused by mutations in the COL17A1 gene is characterized by the frequent development of blisters and erosions on the skin and mucous membranes. The rarity of the disease and the heterogeneity of the underlying mutations renders therapy developments challenging. However, the high number of short in-frame exons facilitates the use of antisense oligonucleotides (AON) to restore collagen 17 (C17) expression by inducing exon skipping. In a personalized approach, we designed and tested three AONs in combination with a cationic liposomal carrier for their ability to induce skipping of COL17A1 exon 7 in 2D culture and in 3D skin equivalents. We show that AON-induced exon skipping excludes the targeted exon from pre-mRNA processing, which restores the reading frame, leading to the expression of a slightly truncated protein. Furthermore, the expression and correct deposition of C17 at the dermal–epidermal junction indicates its functionality. Thus, we assume AON-mediated exon skipping to be a promising tool for the treatment of junctional epidermolysis bullosa, particularly applicable in a personalized manner for rare genotypes.