Research Program for Molecular Therapy of Genodermatoses, EB House Austria, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria.
Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria.
Int J Mol Sci. 2021 Mar 24;22(7):3326. doi: 10.3390/ijms22073326.
Intermediate junctional epidermolysis bullosa caused by mutations in the 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 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.
中间交界型大疱性表皮松解症是由 基因突变引起的，其特征是皮肤和黏膜经常出现水疱和糜烂。这种疾病罕见，潜在突变的异质性使得治疗开发具有挑战性。然而，大量短的内含子外显子使得使用反义寡核苷酸（AON）通过诱导外显子跳跃来恢复胶原蛋白 17（C17）的表达变得可行。在个性化方法中，我们设计并测试了三种 AON 与阳离子脂质体载体结合的能力，以评估它们在 2D 培养和 3D 皮肤等效物中诱导 7 号外显子跳跃的能力。我们表明，AON 诱导的外显子跳跃将靶向外显子排除在 pre-mRNA 加工之外，从而恢复阅读框，导致表达略有截断的蛋白质。此外，C17 在真皮-表皮交界处的表达和正确沉积表明其具有功能性。因此，我们假设 AON 介导的外显子跳跃是治疗交界型大疱性表皮松解症的一种有前途的工具，特别是对于罕见基因型，可以采用个性化的方式应用。
