Antisense-oligonucleotide mediated exon skipping in activin-receptor-like kinase 2: inhibiting the receptor that is overactive in fibrodysplasia ossificans progressiva

Songting Shi; Jie Cai; David J J de Gorter; Gonzalo Sanchez-Duffhues; Dwi U Kemaladewi; Willem M H Hoogaars; Annemieke Aartsma-Rus; Peter A C 't Hoen; Peter ten Dijke

doi:10.1371/journal.pone.0069096

Antisense-oligonucleotide mediated exon skipping in activin-receptor-like kinase 2: inhibiting the receptor that is overactive in fibrodysplasia ossificans progressiva

PLoS One. 2013 Jul 4;8(7):e69096. doi: 10.1371/journal.pone.0069096. Print 2013.

Authors

Songting Shi¹, Jie Cai, David J J de Gorter, Gonzalo Sanchez-Duffhues, Dwi U Kemaladewi, Willem M H Hoogaars, Annemieke Aartsma-Rus, Peter A C 't Hoen, Peter ten Dijke

Affiliation

¹ Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands.

Abstract

Fibrodysplasia ossificans progressiva (FOP) is a rare heritable disease characterized by progressive heterotopic ossification of connective tissues, for which there is presently no definite treatment. A recurrent activating mutation (c.617G→A; R206H) of activin receptor-like kinase 2 (ACVR1/ALK2), a BMP type I receptor, has been shown as the main cause of FOP. This mutation constitutively activates the BMP signaling pathway and initiates the formation of heterotopic bone. In this study, we have designed antisense oligonucleotides (AONs) to knockdown mouse ALK2 expression by means of exon skipping. The ALK2 AON could induce exon skipping in cells, which was accompanied by decreased ALK2 mRNA levels and impaired BMP signaling. In addition, the ALK2 AON potentiated muscle differentiation and repressed BMP6-induced osteoblast differentiation. Our results therefore provide a potential therapeutic approach for the treatment of FOP disease by reducing the excessive ALK2 activity in FOP patients.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Activin Receptors, Type I / antagonists & inhibitors*
Activin Receptors, Type I / genetics
Activin Receptors, Type I / metabolism
Animals
Bone Morphogenetic Protein 6 / genetics
Bone Morphogenetic Protein 6 / metabolism
Cell Differentiation
Exons*
Gene Expression Regulation
Gene Knockdown Techniques
Genetic Therapy / methods
Humans
Mice
Muscle Cells / cytology*
Muscle Cells / metabolism
Mutation
Myoblasts / cytology*
Myoblasts / metabolism
Myositis Ossificans / genetics
Myositis Ossificans / metabolism
Myositis Ossificans / pathology
Myositis Ossificans / therapy*
Oligonucleotides, Antisense / chemical synthesis
Oligonucleotides, Antisense / genetics*
Osteoblasts / metabolism
Osteoblasts / pathology
Osteogenesis / genetics
Signal Transduction

Substances

Bone Morphogenetic Protein 6
Oligonucleotides, Antisense
Activin Receptors, Type I
Acvr1 protein, mouse

Grants and funding

This study was supported by Dutch Ministry for Economic Affairs (IOP Genomics grant IGE7001) “Towards broad clinical and technological application of gene expression engineering by exon skipping”, Netherlands Research Council (NWO-MW), Cancer Genomics Centre Netherlands and Centre for Biomedical Genetics; and by funds from LeDucq foundation and AO Start-up Grant S-12-27S: Targeting Endothelial-to-Mesenchymal transition in Fibrodysplasia Ossificans Progressiva. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.