A recurrent COL6A1 pseudoexon insertion causes muscular dystrophy and is effectively targeted by splice-correction therapies

The clinical application of advanced next-generation sequencing technologies is increasingly uncovering novel classes of mutations that may serve as potential targets for precision medicine therapeutics. Here, we show that a deep intronic splice defect in the COL6A1 gene, originally discovered by ap...

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Bibliographic Details
Published in:	JCI insight 2019-03, Vol.4 (6)
Main Authors:	Bolduc, Véronique, Foley, A Reghan, Solomon-Degefa, Herimela, Sarathy, Apurva, Donkervoort, Sandra, Hu, Ying, Chen, Grace S, Sizov, Katherine, Nalls, Matthew, Zhou, Haiyan, Aguti, Sara, Cummings, Beryl B, Lek, Monkol, Tukiainen, Taru, Marshall, Jamie L, Regev, Oded, Marek-Yagel, Dina, Sarkozy, Anna, Butterfield, Russell J, Jou, Cristina, Jimenez-Mallebrera, Cecilia, Li, Yan, Gartioux, Corine, Mamchaoui, Kamel, Allamand, Valérie, Gualandi, Francesca, Ferlini, Alessandra, Hanssen, Eric, Wilton, Steve D, Lamandé, Shireen R, MacArthur, Daniel G, Wagener, Raimund, Muntoni, Francesco, Bönnemann, Carsten G
Format:	Article
Language:	eng
Subjects:	Base Sequence Collagen Type VI - genetics CRISPR-Cas Systems DNA Mutational Analysis Exons - genetics Fibroblasts - metabolism Fibroblasts - pathology Gene Expression Genetic Predisposition to Disease - genetics Genetic Therapy - methods High-Throughput Nucleotide Sequencing Human health and pathology Humans Introns - genetics Life Sciences Muscular Dystrophies - genetics Muscular Dystrophies - therapy Mutation RNA Splice Sites RNA Splicing RNA, Messenger - metabolism Skin - pathology Tissues and Organs
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Summary:	The clinical application of advanced next-generation sequencing technologies is increasingly uncovering novel classes of mutations that may serve as potential targets for precision medicine therapeutics. Here, we show that a deep intronic splice defect in the COL6A1 gene, originally discovered by applying muscle RNA sequencing in patients with clinical findings of collagen VI-related dystrophy (COL6-RD), inserts an in-frame pseudoexon into COL6A1 mRNA, encodes a mutant collagen α1(VI) protein that exerts a dominant-negative effect on collagen VI matrix assembly, and provides a unique opportunity for splice-correction approaches aimed at restoring normal gene expression. Using splice-modulating antisense oligomers, we efficiently skipped the pseudoexon in patient-derived fibroblast cultures and restored a wild-type matrix. Similarly, we used CRISPR/Cas9 to precisely delete an intronic sequence containing the pseudoexon and efficiently abolish its inclusion while preserving wild-type splicing. Considering that this splice defect is emerging as one of the single most frequent mutations in COL6-RD, the design of specific and effective splice-correction therapies offers a promising path for clinical translation.
ISSN:	2379-3708 2379-3708