Abstract
Fibrodysplasia ossificans progressiva (FOP) is a rare autosomal-dominant disorder characterized by progressive heterotopic ossification. More than 95% of cases are caused by a recurrent mutation (617G>A; R206H) of ACVR1/ALK2, a bone morphogenetic protein (BMP) type I receptor. Recent studies revealed that ACVR1R206H induces heterotopic ossification by aberrant activation in response to activin A. Because ACVR1R206H is a hyperactive receptor, a promising therapeutic strategy is to decrease the activity of ACVR1 in patients. Here, we describe a method to reduce ACVR1 expression in FOP patient cells by exon skipping in ACVR1 mRNAs using phosphorodiamidate morpholino oligomers (PMOs). This strategy can be applied to the screen to select antisense oligomers to knockdown not only ACVR1 but also genes which cause other autosomal-dominant genetic diseases.
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References
Pignolo RJ, Shore EM, Kaplan FS (2011) Fibrodysplasia ossificans progressiva: clinical and genetic aspects. Orphanet J Rare Dis 6:80. https://doi.org/10.1186/1750-1172-6-80
Chakkalakal SA, Uchibe K, Convente MR et al (2016) Palovarotene inhibits heterotopic ossification and maintains limb mobility and growth in mice with the human ACVR1(R206H) fibrodysplasia ossificans progressiva (FOP) mutation. J Bone Miner Res 31(9):1666–1675. https://doi.org/10.1002/jbmr.2820
Hino K, Horigome K, Nishio M et al (2017) Activin-a enhances mTOR signaling to promote aberrant chondrogenesis in fibrodysplasia ossificans progressiva. J Clin Invest 127(9):3339–3352. https://doi.org/10.1172/JCI93521
Shore EM, Xu M, Feldman GJ et al (2006) A recurrent mutation in the BMP type I receptor ACVR1 causes inherited and sporadic fibrodysplasia ossificans progressiva. Nat Genet 38(5):525–527. https://doi.org/10.1038/ng1783
Kaplan FS, Xu M, Seemann P et al (2009) Classic and atypical fibrodysplasia ossificans progressiva (FOP) phenotypes are caused by mutations in the bone morphogenetic protein (BMP) type I receptor ACVR1. Hum Mutat 30(3):379–390. https://doi.org/10.1002/humu.20868
Hatsell SJ, Idone V, Wolken DM et al (2015) ACVR1R206H receptor mutation causes fibrodysplasia ossificans progressiva by imparting responsiveness to activin a. Sci Transl Med 7(303):303ra137. https://doi.org/10.1126/scitranslmed.aac4358
Hino K, Ikeya M, Horigome K et al (2015) Neofunction of ACVR1 in fibrodysplasia ossificans progressiva. Proc Natl Acad Sci U S A 112(50):15438–15443. https://doi.org/10.1073/pnas.1510540112
Dey D, Bagarova J, Hatsell SJ et al (2016) Two tissue-resident progenitor lineages drive distinct phenotypes of heterotopic ossification. Sci Transl Med 8(366):366ra163. https://doi.org/10.1126/scitranslmed.aaf1090
Buczkowicz P, Hoeman C, Rakopoulos P et al (2014) Genomic analysis of diffuse intrinsic pontine gliomas identifies three molecular subgroups and recurrent activating ACVR1 mutations. Nat Genet 46(5):451–456. https://doi.org/10.1038/ng.2936
Taylor KR, Mackay A, Truffaux N et al (2014) Recurrent activating ACVR1 mutations in diffuse intrinsic pontine glioma. Nat Genet 46(5):457–461. https://doi.org/10.1038/ng.2925
Wu G, Diaz AK, Paugh BS et al (2014) The genomic landscape of diffuse intrinsic pontine glioma and pediatric non-brainstem high-grade glioma. Nat Genet 46(5):444–450. https://doi.org/10.1038/ng.2938
Lee J, Yokota T (2016) Translational research in nucleic acid therapies for muscular dystrophies. In: Takeda S, Miyagoe-Suzuki Y, Mori-Yoshimura M (eds) Translational research in muscular dystrophy. Springer Japan, Tokyo, Japan, pp 87–102. https://doi.org/10.1007/978-4-431-55678-7_6
Lim KR, Maruyama R, Yokota T (2017) Eteplirsen in the treatment of Duchenne muscular dystrophy. Drug Des Devel Ther 11:533–545. https://doi.org/10.2147/DDDT.S97635
Shi S, Cai J, de Gorter DJ et al (2013) Antisense-oligonucleotide mediated exon skipping in activin-receptor-like kinase 2: inhibiting the receptor that is overactive in fibrodysplasia ossificans progressiva. PLoS One 8(7):e69096. https://doi.org/10.1371/journal.pone.0069096
Acknowledgments
This work was supported by the University of Alberta Faculty of Medicine and Dentistry, International FOP Association/The Canadian FOP Network Research Grant, Gilbert K. Winter Funds, the Canadian Institutes of Health Research (CIHR), the Friends of Garrett Cumming Research Funds, HM Toupin Neurological Science Research Funds, the Muscular Dystrophy Canada, the Canada Foundation for Innovation, Alberta Enterprise and Advanced Education, Rare Disease Foundation/BC Children’s Hospital Foundation Microgrant, and the Women and Children’s Health Research Institute (WCHRI).
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Maruyama, R., Yokota, T. (2018). Morpholino-Mediated Exon Skipping Targeting Human ACVR1/ALK2 for Fibrodysplasia Ossificans Progressiva. In: Yokota, T., Maruyama, R. (eds) Exon Skipping and Inclusion Therapies. Methods in Molecular Biology, vol 1828. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8651-4_32
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DOI: https://doi.org/10.1007/978-1-4939-8651-4_32
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