Somatic Therapy of a Mouse SMA Model with a U7 SnRNA Gene Correcting SMN2 Splicing

Overview

Journal Mol Ther

Publisher Cell Press

Specialties Molecular Biology
Pharmacology

Date 2016 Jul 27

PMID 27456062

Citations 12

Authors

Philipp Odermatt

Judith Trub

Lavinia Furrer

Roger Fricker

Andreas Marti

Daniel Schumperli

Affiliations

Soon will be listed here.

Abstract

Spinal Muscular Atrophy is due to the loss of SMN1 gene function. The duplicate gene SMN2 produces some, but not enough, SMN protein because most transcripts lack exon 7. Thus, promoting the inclusion of this exon is a therapeutic option. We show that a somatic gene therapy using the gene for a modified U7 RNA which stimulates this splicing has a profound and persistent therapeutic effect on the phenotype of a severe Spinal Muscular Atrophy mouse model. To this end, the U7 gene and vector and the production of pure, highly concentrated self-complementary (sc) adenovirus-associated virus 9 vector particles were optimized. Introduction of the functional vector into motoneurons of newborn Spinal Muscular Atrophy mice by intracerebroventricular injection led to a highly significant, dose-dependent increase in life span and improvement of muscle functions. Besides the central nervous system, the therapeutic U7 RNA was expressed in the heart and liver which may additionally have contributed to the observed therapeutic efficacy. This approach provides an additional therapeutic option for Spinal Muscular Atrophy and could also be adapted to treat other diseases of the central nervous system with regulatory small RNA genes.

Citing Articles

Design and evaluation of antisense sequence length for modified mouse U7 small nuclear RNA to induce efficient pre-messenger RNA splicing modulation in vitro.

Shimo T, Ueda O, Yamamoto S PLoS One. 2024; 19(7):e0305012.

PMID: 38980892 PMC: 11232981. DOI: 10.1371/journal.pone.0305012.

RNA-Based Therapeutic Technology.

Mashima R, Takada S, Miyamoto Y Int J Mol Sci. 2023; 24(20).

PMID: 37894911 PMC: 10607345. DOI: 10.3390/ijms242015230.

Systematic review and meta-analysis determining the benefits of in vivo genetic therapy in spinal muscular atrophy rodent models.

Chilcott E, Muiruri E, Hirst T, Yanez-Munoz R Gene Ther. 2021; 29(9):498-512.

PMID: 34611322 PMC: 9482879. DOI: 10.1038/s41434-021-00292-4.

U7 snRNA: A tool for gene therapy.

Gadgil A, Raczynska K J Gene Med. 2021; 23(4):e3321.

PMID: 33590603 PMC: 8243935. DOI: 10.1002/jgm.3321.

Lack of Toxicity in Nonhuman Primates Receiving Clinically Relevant Doses of an AAV9.U7snRNA Vector Designed to Induce Exon 2 Skipping.

Gushchina L, Frair E, Rohan N, J Bradley A, Simmons T, Chavan H Hum Gene Ther. 2021; 32(17-18):882-894.

PMID: 33406986 PMC: 10112461. DOI: 10.1089/hum.2020.286.

References

Marquis J, Kampfer S, Angehrn L, Schumperli D . Doxycycline-controlled splicing modulation by regulated antisense U7 snRNA expression cassettes. Gene Ther. 2008; 16(1):70-7. DOI: 10.1038/gt.2008.138. View

Hua Y, Vickers T, Okunola H, Bennett C, Krainer A . Antisense masking of an hnRNP A1/A2 intronic splicing silencer corrects SMN2 splicing in transgenic mice. Am J Hum Genet. 2008; 82(4):834-48. PMC: 2427210. DOI: 10.1016/j.ajhg.2008.01.014. View

Gruber A, Soldati D, Burri M, Schumperli D . Isolation of an active gene and of two pseudogenes for mouse U7 small nuclear RNA. Biochim Biophys Acta. 1991; 1088(1):151-4. DOI: 10.1016/0167-4781(91)90167-k. View

Goyenvalle A, Babbs A, Wright J, Wilkins V, Powell D, Garcia L . Rescue of severely affected dystrophin/utrophin-deficient mice through scAAV-U7snRNA-mediated exon skipping. Hum Mol Genet. 2012; 21(11):2559-71. PMC: 3349427. DOI: 10.1093/hmg/dds082. View

Haurigot V, Marco S, Ribera A, Garcia M, Ruzo A, Villacampa P . Whole body correction of mucopolysaccharidosis IIIA by intracerebrospinal fluid gene therapy. J Clin Invest. 2013; 123(8):3254-3271. PMC: 3726158. DOI: 10.1172/JCI66778. View

Kashima T, Rao N, David C, Manley J . hnRNP A1 functions with specificity in repression of SMN2 exon 7 splicing. Hum Mol Genet. 2007; 16(24):3149-59. DOI: 10.1093/hmg/ddm276. View

Hua Y, Liu Y, Sahashi K, Rigo F, Bennett C, Krainer A . Motor neuron cell-nonautonomous rescue of spinal muscular atrophy phenotypes in mild and severe transgenic mouse models. Genes Dev. 2015; 29(3):288-97. PMC: 4318145. DOI: 10.1101/gad.256644.114. View

Raczynska K, Ruepp M, Brzek A, Reber S, Romeo V, Rindlisbacher B . FUS/TLS contributes to replication-dependent histone gene expression by interaction with U7 snRNPs and histone-specific transcription factors. Nucleic Acids Res. 2015; 43(20):9711-28. PMC: 4787759. DOI: 10.1093/nar/gkv794. View

Meyer K, Ferraiuolo L, Schmelzer L, Braun L, McGovern V, Likhite S . Improving single injection CSF delivery of AAV9-mediated gene therapy for SMA: a dose-response study in mice and nonhuman primates. Mol Ther. 2014; 23(3):477-87. PMC: 4351452. DOI: 10.1038/mt.2014.210. View

10.

Crawford T, Pardo C . The neurobiology of childhood spinal muscular atrophy. Neurobiol Dis. 1996; 3(2):97-110. DOI: 10.1006/nbdi.1996.0010. View

11.

Rudnik-Schoneborn S, Heller R, Berg C, Betzler C, Grimm T, Eggermann T . Congenital heart disease is a feature of severe infantile spinal muscular atrophy. J Med Genet. 2008; 45(10):635-8. DOI: 10.1136/jmg.2008.057950. View

12.

Butchbach M, Edwards J, Burghes A . Abnormal motor phenotype in the SMNDelta7 mouse model of spinal muscular atrophy. Neurobiol Dis. 2007; 27(2):207-19. PMC: 2700002. DOI: 10.1016/j.nbd.2007.04.009. View

13.

Wu Z, Yang H, Colosi P . Effect of genome size on AAV vector packaging. Mol Ther. 2009; 18(1):80-6. PMC: 2839202. DOI: 10.1038/mt.2009.255. View

14.

Gil-Farina I, Fronza R, Kaeppel C, Lopez-Franco E, Ferreira V, DAvola D . Recombinant AAV Integration Is Not Associated With Hepatic Genotoxicity in Nonhuman Primates and Patients. Mol Ther. 2016; 24(6):1100-1105. PMC: 4923321. DOI: 10.1038/mt.2016.52. View

15.

Le T, Pham L, Butchbach M, Zhang H, Monani U, Coovert D . SMNDelta7, the major product of the centromeric survival motor neuron (SMN2) gene, extends survival in mice with spinal muscular atrophy and associates with full-length SMN. Hum Mol Genet. 2005; 14(6):845-57. DOI: 10.1093/hmg/ddi078. View

16.

Liu H, Shafey D, Moores J, Kothary R . Neurodevelopmental consequences of Smn depletion in a mouse model of spinal muscular atrophy. J Neurosci Res. 2009; 88(1):111-22. DOI: 10.1002/jnr.22189. View

17.

Duque S, Arnold W, Odermatt P, Li X, Porensky P, Schmelzer L . A large animal model of spinal muscular atrophy and correction of phenotype. Ann Neurol. 2014; 77(3):399-414. PMC: 4453930. DOI: 10.1002/ana.24332. View

18.

McGovern V, Iyer C, Arnold W, Gombash S, Zaworski P, Blatnik 3rd A . SMN expression is required in motor neurons to rescue electrophysiological deficits in the SMNΔ7 mouse model of SMA. Hum Mol Genet. 2015; 24(19):5524-41. PMC: 4572068. DOI: 10.1093/hmg/ddv283. View

19.

Bell P, Wang L, Lebherz C, Flieder D, Bove M, Wu D . No evidence for tumorigenesis of AAV vectors in a large-scale study in mice. Mol Ther. 2005; 12(2):299-306. DOI: 10.1016/j.ymthe.2005.03.020. View

20.

Zincarelli C, Soltys S, Rengo G, Rabinowitz J . Analysis of AAV serotypes 1-9 mediated gene expression and tropism in mice after systemic injection. Mol Ther. 2008; 16(6):1073-80. DOI: 10.1038/mt.2008.76. View