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

TitleA recurrent COL6A1 pseudoexon insertion causes muscular dystrophy and is effectively targeted by splice-correction therapies.
Publication TypeJournal Article
Year of Publication2019
AuthorsBolduc, V, A Foley, R, Solomon-Degefa, H, Sarathy, A, Donkervoort, S, Hu, Y, Chen, GS, Sizov, K, Nalls, M, Zhou, H, Aguti, S, Cummings, BB, Lek, M, Tukiainen, T, Marshall, JL, Regev, O, Marek-Yagel, D, Sarkozy, A, Butterfield, RJ, Jou, C, Jimenez-Mallebrera, C, Li, Y, Gartioux, C, Mamchaoui, K, Allamand, V, Gualandi, F, Ferlini, A, Hanssen, E, Wilton, SD, Lamandé, SR, MacArthur, DG, Wagener, R, Muntoni, F, Bönnemann, CG
Corporate AuthorsCOL6A1 Intron 11 Study Group
JournalJCI Insight
Volume4
Issue6
Date Published2019 Mar 21
ISSN2379-3708
Abstract

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.

DOI10.1172/jci.insight.124403
Alternate JournalJCI Insight
PubMed ID30895940
PubMed Central IDPMC6483063
Grant ListK08 NS097631 / NS / NINDS NIH HHS / United States
UM1 HG008900 / HG / NHGRI NIH HHS / United States
ZIA NS003129 / NS / NINDS NIH HHS / United States