MRI roadmap-guided transendocardial delivery of exon-skipping recombinant adeno-associated virus restores dystrophin expression in a canine model of Duchenne muscular dystrophy

I. M. Barbash; S. Cecchini; A. Z. Faranesh; T. Virag; L. Li; Y. Yang; R. F. Hoyt; J. N. Kornegay; J. R. Bogan; L. Garcia; R. J. Lederman; R. M. Kotin

doi:10.1038/gt.2012.38

MRI roadmap-guided transendocardial delivery of exon-skipping recombinant adeno-associated virus restores dystrophin expression in a canine model of Duchenne muscular dystrophy

I. M. Barbash, S. Cecchini, A. Z. Faranesh, T. Virag, L. Li, Y. Yang, R. F. Hoyt, J. N. Kornegay, J. R. Bogan, L. Garcia, R. J. Lederman, R. M. Kotin^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Duchenne muscular dystrophy (DMD) cardiomyopathy patients currently have no therapeutic options. We evaluated catheter-based transendocardial delivery of a recombinant adeno-associated virus (rAAV) expressing a small nuclear U7 RNA (U7smOPT) complementary to specific cis-acting splicing signals. Eliminating specific exons restores the open reading frame resulting in translation of truncated dystrophin protein. To test this approach in a clinically relevant DMD model, golden retriever muscular dystrophy (GRMD) dogs received serotype 6 rAAV-U7smOPT via the intracoronary or transendocardial route. Transendocardial injections were administered with an injection-tipped catheter and fluoroscopic guidance using X-ray fused with magnetic resonance imaging (XFM) roadmaps. Three months after treatment, tissues were analyzed for DNA, RNA, dystrophin protein, and histology. Whereas intracoronary delivery did not result in effective transduction, transendocardial injections, XFM guidance, enabled 30±10 non-overlapping injections per animal. Vector DNA was detectable in all samples tested and ranged from <1 to >3000 vector genome copies per cell. RNA analysis, western blot analysis, and immunohistology demonstrated extensive expression of skipped RNA and dystrophin protein in the treated myocardium. Left ventricular function remained unchanged over a 3-month follow-up. These results demonstrated that effective transendocardial delivery of rAAV-U7smOPT was achieved using XFM. This approach restores an open reading frame for dystrophin in affected dogs and has potential clinical utility.

Original language	English
Pages (from-to)	274-282
Number of pages	9
Journal	Gene Therapy
Volume	20
Issue number	3
DOIs	https://doi.org/10.1038/gt.2012.38
State	Published - Mar 2013
Externally published	Yes

Access to Document

10.1038/gt.2012.38

Cite this

Barbash, I. M., Cecchini, S., Faranesh, A. Z., Virag, T., Li, L., Yang, Y., Hoyt, R. F., Kornegay, J. N., Bogan, J. R., Garcia, L., Lederman, R. J., & Kotin, R. M. (2013). MRI roadmap-guided transendocardial delivery of exon-skipping recombinant adeno-associated virus restores dystrophin expression in a canine model of Duchenne muscular dystrophy. Gene Therapy, 20(3), 274-282. https://doi.org/10.1038/gt.2012.38

@article{22282af357144efabf00cf75afbae2b4,

title = "MRI roadmap-guided transendocardial delivery of exon-skipping recombinant adeno-associated virus restores dystrophin expression in a canine model of Duchenne muscular dystrophy",

abstract = "Duchenne muscular dystrophy (DMD) cardiomyopathy patients currently have no therapeutic options. We evaluated catheter-based transendocardial delivery of a recombinant adeno-associated virus (rAAV) expressing a small nuclear U7 RNA (U7smOPT) complementary to specific cis-acting splicing signals. Eliminating specific exons restores the open reading frame resulting in translation of truncated dystrophin protein. To test this approach in a clinically relevant DMD model, golden retriever muscular dystrophy (GRMD) dogs received serotype 6 rAAV-U7smOPT via the intracoronary or transendocardial route. Transendocardial injections were administered with an injection-tipped catheter and fluoroscopic guidance using X-ray fused with magnetic resonance imaging (XFM) roadmaps. Three months after treatment, tissues were analyzed for DNA, RNA, dystrophin protein, and histology. Whereas intracoronary delivery did not result in effective transduction, transendocardial injections, XFM guidance, enabled 30±10 non-overlapping injections per animal. Vector DNA was detectable in all samples tested and ranged from <1 to >3000 vector genome copies per cell. RNA analysis, western blot analysis, and immunohistology demonstrated extensive expression of skipped RNA and dystrophin protein in the treated myocardium. Left ventricular function remained unchanged over a 3-month follow-up. These results demonstrated that effective transendocardial delivery of rAAV-U7smOPT was achieved using XFM. This approach restores an open reading frame for dystrophin in affected dogs and has potential clinical utility.",

author = "Barbash, {I. M.} and S. Cecchini and Faranesh, {A. Z.} and T. Virag and L. Li and Y. Yang and Hoyt, {R. F.} and Kornegay, {J. N.} and Bogan, {J. R.} and L. Garcia and Lederman, {R. J.} and Kotin, {R. M.}",

note = "Funding Information: We are grateful to the NHLBI Laboratory of Animal Medicine and Surgery technologists for assistance with animal experiments, and the veterinarians and technicians of the Division of Veterinary Research and Victor Wright for assistance with MRI scans. We appreciate the professional skills and advice of the Dr Christian Combs, NHLBI Light Microscopy Core, and Dr Xu-Zi Yu, NHLBI Pathology Core. Dr Victoria Joan Hoffman of Diagnostic & Research Services Branch, Division of Veterinary Resources, NIH, provided essential support and advice. Dr H Lee Sweeney provided helpful comments and discussion. We thank Boston Scientific for providing the injection catheters. Funding was provided by the National Heart, Lung, and Blood Institute, Division of Intramural Research, and the International Collaborative Effort (ICE) for Duchenne Muscular Dystrophy. The ICE consists of the Duchenne Parent Project France and the Association Monagasque Contre les Myopathies.",

year = "2013",

month = mar,

doi = "10.1038/gt.2012.38",

language = "אנגלית",

volume = "20",

pages = "274--282",

journal = "Gene Therapy",

issn = "0969-7128",

publisher = "Nature Publishing Group",

number = "3",

}

Barbash, IM, Cecchini, S, Faranesh, AZ, Virag, T, Li, L, Yang, Y, Hoyt, RF, Kornegay, JN, Bogan, JR, Garcia, L, Lederman, RJ & Kotin, RM 2013, 'MRI roadmap-guided transendocardial delivery of exon-skipping recombinant adeno-associated virus restores dystrophin expression in a canine model of Duchenne muscular dystrophy', Gene Therapy, vol. 20, no. 3, pp. 274-282. https://doi.org/10.1038/gt.2012.38

TY - JOUR

T1 - MRI roadmap-guided transendocardial delivery of exon-skipping recombinant adeno-associated virus restores dystrophin expression in a canine model of Duchenne muscular dystrophy

AU - Barbash, I. M.

AU - Cecchini, S.

AU - Faranesh, A. Z.

AU - Virag, T.

AU - Li, L.

AU - Yang, Y.

AU - Hoyt, R. F.

AU - Kornegay, J. N.

AU - Bogan, J. R.

AU - Garcia, L.

AU - Lederman, R. J.

AU - Kotin, R. M.

N1 - Funding Information: We are grateful to the NHLBI Laboratory of Animal Medicine and Surgery technologists for assistance with animal experiments, and the veterinarians and technicians of the Division of Veterinary Research and Victor Wright for assistance with MRI scans. We appreciate the professional skills and advice of the Dr Christian Combs, NHLBI Light Microscopy Core, and Dr Xu-Zi Yu, NHLBI Pathology Core. Dr Victoria Joan Hoffman of Diagnostic & Research Services Branch, Division of Veterinary Resources, NIH, provided essential support and advice. Dr H Lee Sweeney provided helpful comments and discussion. We thank Boston Scientific for providing the injection catheters. Funding was provided by the National Heart, Lung, and Blood Institute, Division of Intramural Research, and the International Collaborative Effort (ICE) for Duchenne Muscular Dystrophy. The ICE consists of the Duchenne Parent Project France and the Association Monagasque Contre les Myopathies.

PY - 2013/3

Y1 - 2013/3

N2 - Duchenne muscular dystrophy (DMD) cardiomyopathy patients currently have no therapeutic options. We evaluated catheter-based transendocardial delivery of a recombinant adeno-associated virus (rAAV) expressing a small nuclear U7 RNA (U7smOPT) complementary to specific cis-acting splicing signals. Eliminating specific exons restores the open reading frame resulting in translation of truncated dystrophin protein. To test this approach in a clinically relevant DMD model, golden retriever muscular dystrophy (GRMD) dogs received serotype 6 rAAV-U7smOPT via the intracoronary or transendocardial route. Transendocardial injections were administered with an injection-tipped catheter and fluoroscopic guidance using X-ray fused with magnetic resonance imaging (XFM) roadmaps. Three months after treatment, tissues were analyzed for DNA, RNA, dystrophin protein, and histology. Whereas intracoronary delivery did not result in effective transduction, transendocardial injections, XFM guidance, enabled 30±10 non-overlapping injections per animal. Vector DNA was detectable in all samples tested and ranged from <1 to >3000 vector genome copies per cell. RNA analysis, western blot analysis, and immunohistology demonstrated extensive expression of skipped RNA and dystrophin protein in the treated myocardium. Left ventricular function remained unchanged over a 3-month follow-up. These results demonstrated that effective transendocardial delivery of rAAV-U7smOPT was achieved using XFM. This approach restores an open reading frame for dystrophin in affected dogs and has potential clinical utility.

AB - Duchenne muscular dystrophy (DMD) cardiomyopathy patients currently have no therapeutic options. We evaluated catheter-based transendocardial delivery of a recombinant adeno-associated virus (rAAV) expressing a small nuclear U7 RNA (U7smOPT) complementary to specific cis-acting splicing signals. Eliminating specific exons restores the open reading frame resulting in translation of truncated dystrophin protein. To test this approach in a clinically relevant DMD model, golden retriever muscular dystrophy (GRMD) dogs received serotype 6 rAAV-U7smOPT via the intracoronary or transendocardial route. Transendocardial injections were administered with an injection-tipped catheter and fluoroscopic guidance using X-ray fused with magnetic resonance imaging (XFM) roadmaps. Three months after treatment, tissues were analyzed for DNA, RNA, dystrophin protein, and histology. Whereas intracoronary delivery did not result in effective transduction, transendocardial injections, XFM guidance, enabled 30±10 non-overlapping injections per animal. Vector DNA was detectable in all samples tested and ranged from <1 to >3000 vector genome copies per cell. RNA analysis, western blot analysis, and immunohistology demonstrated extensive expression of skipped RNA and dystrophin protein in the treated myocardium. Left ventricular function remained unchanged over a 3-month follow-up. These results demonstrated that effective transendocardial delivery of rAAV-U7smOPT was achieved using XFM. This approach restores an open reading frame for dystrophin in affected dogs and has potential clinical utility.

UR - http://www.scopus.com/inward/record.url?scp=84874786215&partnerID=8YFLogxK

U2 - 10.1038/gt.2012.38

DO - 10.1038/gt.2012.38

M3 - מאמר

C2 - 22551778

AN - SCOPUS:84874786215

VL - 20

SP - 274

EP - 282

JO - Gene Therapy

JF - Gene Therapy

SN - 0969-7128

IS - 3

ER -

MRI roadmap-guided transendocardial delivery of exon-skipping recombinant adeno-associated virus restores dystrophin expression in a canine model of Duchenne muscular dystrophy

Abstract

Access to Document

Other files and links

Fingerprint

Cite this