Systemic delivery of AAV5, AAV8, and AAV9 packaging a C5-12-microdystrophin-FLAG expression cassette in non-human primates
Safely achieving therapeutic expression levels with adeno-associated virus (AAV) gene therapy is a significant challenge for treating the large muscle mass in humans. Non-human primates (NHPs) provide a more accurate assessment of the feasibility of achieving an effective and safe dose than rodents....
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| Language: | English |
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Elsevier
2025-03-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050125000063 |
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| author | Mengping Liu Erica Cook Yanshan Dai Erich Ehlert Francois du Plessis Jacek Lubelski Bogdan G. Sleczka Petia Shipkova Zhuyin Li Joshua Gamse David Gordon Leonard P. Adam Paul C. Levesque Glen B. Banks |
| author_facet | Mengping Liu Erica Cook Yanshan Dai Erich Ehlert Francois du Plessis Jacek Lubelski Bogdan G. Sleczka Petia Shipkova Zhuyin Li Joshua Gamse David Gordon Leonard P. Adam Paul C. Levesque Glen B. Banks |
| author_sort | Mengping Liu |
| collection | DOAJ |
| description | Safely achieving therapeutic expression levels with adeno-associated virus (AAV) gene therapy is a significant challenge for treating the large muscle mass in humans. Non-human primates (NHPs) provide a more accurate assessment of the feasibility of achieving an effective and safe dose than rodents. Here, we compared a single systemic administration of AAV5, AAV8, or AAV9 in NHPs, each packaging the C5-12-microdystrophin-FLAG expression cassette. At 1 month post-dose, we compared tissue vector genomes, mRNA, and microdystrophin-FLAG protein levels by meso-scale discovery-enzyme-linked immunosorbent assay, liquid chromatography-mass spectrometry, and immunofluorescence. The C5-12 promoter was highly selective for heart and skeletal muscles, when compared to off-target tissues such as peripheral blood mononuclear cells, lung, liver, and kidney. AAV8 led to higher levels of microdystrophin-FLAG mRNA and protein in the cardiac ventricles and skeletal muscles when compared to AAV5 or AAV9. The AAV8-microdystrophin-FLAG led to ∼20% of wild-type NHP dystrophin protein expression levels and was located on the sarcolemma of ∼40% of skeletal muscles fibers and ∼15% of left ventricular cardiomyocytes. Hematology, serum chemistry, and pathology were unremarkable. Thus, a systemic dose of ∼1.18 × 1014 vector genomes/kg AAV8 is predicted to be safe and efficacious for treating Duchenne muscular dystrophy (DMD) but has significant room for improvement. |
| format | Article |
| id | doaj-art-d663094d0bda41b49e3aaa82e7a23c9e |
| institution | Kabale University |
| issn | 2329-0501 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj-art-d663094d0bda41b49e3aaa82e7a23c9e2025-02-12T05:31:17ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012025-03-01331101411Systemic delivery of AAV5, AAV8, and AAV9 packaging a C5-12-microdystrophin-FLAG expression cassette in non-human primatesMengping Liu0Erica Cook1Yanshan Dai2Erich Ehlert3Francois du Plessis4Jacek Lubelski5Bogdan G. Sleczka6Petia Shipkova7Zhuyin Li8Joshua Gamse9David Gordon10Leonard P. Adam11Paul C. Levesque12Glen B. Banks13Bristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USABristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USABristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USAuniQure, Paasheuvelweg 25a, 1105 BP Amsterdam, the NetherlandsuniQure, Paasheuvelweg 25a, 1105 BP Amsterdam, the NetherlandsuniQure, Paasheuvelweg 25a, 1105 BP Amsterdam, the NetherlandsBristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USABristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USABristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USABristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USABristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USABristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USABristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USABristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USA; Corresponding author: Glen B. Banks, Bristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USA.Safely achieving therapeutic expression levels with adeno-associated virus (AAV) gene therapy is a significant challenge for treating the large muscle mass in humans. Non-human primates (NHPs) provide a more accurate assessment of the feasibility of achieving an effective and safe dose than rodents. Here, we compared a single systemic administration of AAV5, AAV8, or AAV9 in NHPs, each packaging the C5-12-microdystrophin-FLAG expression cassette. At 1 month post-dose, we compared tissue vector genomes, mRNA, and microdystrophin-FLAG protein levels by meso-scale discovery-enzyme-linked immunosorbent assay, liquid chromatography-mass spectrometry, and immunofluorescence. The C5-12 promoter was highly selective for heart and skeletal muscles, when compared to off-target tissues such as peripheral blood mononuclear cells, lung, liver, and kidney. AAV8 led to higher levels of microdystrophin-FLAG mRNA and protein in the cardiac ventricles and skeletal muscles when compared to AAV5 or AAV9. The AAV8-microdystrophin-FLAG led to ∼20% of wild-type NHP dystrophin protein expression levels and was located on the sarcolemma of ∼40% of skeletal muscles fibers and ∼15% of left ventricular cardiomyocytes. Hematology, serum chemistry, and pathology were unremarkable. Thus, a systemic dose of ∼1.18 × 1014 vector genomes/kg AAV8 is predicted to be safe and efficacious for treating Duchenne muscular dystrophy (DMD) but has significant room for improvement.http://www.sciencedirect.com/science/article/pii/S2329050125000063adeno-associated viruspromotersystemic deliverynon-human primatedystrophinAAV |
| spellingShingle | Mengping Liu Erica Cook Yanshan Dai Erich Ehlert Francois du Plessis Jacek Lubelski Bogdan G. Sleczka Petia Shipkova Zhuyin Li Joshua Gamse David Gordon Leonard P. Adam Paul C. Levesque Glen B. Banks Systemic delivery of AAV5, AAV8, and AAV9 packaging a C5-12-microdystrophin-FLAG expression cassette in non-human primates Molecular Therapy: Methods & Clinical Development adeno-associated virus promoter systemic delivery non-human primate dystrophin AAV |
| title | Systemic delivery of AAV5, AAV8, and AAV9 packaging a C5-12-microdystrophin-FLAG expression cassette in non-human primates |
| title_full | Systemic delivery of AAV5, AAV8, and AAV9 packaging a C5-12-microdystrophin-FLAG expression cassette in non-human primates |
| title_fullStr | Systemic delivery of AAV5, AAV8, and AAV9 packaging a C5-12-microdystrophin-FLAG expression cassette in non-human primates |
| title_full_unstemmed | Systemic delivery of AAV5, AAV8, and AAV9 packaging a C5-12-microdystrophin-FLAG expression cassette in non-human primates |
| title_short | Systemic delivery of AAV5, AAV8, and AAV9 packaging a C5-12-microdystrophin-FLAG expression cassette in non-human primates |
| title_sort | systemic delivery of aav5 aav8 and aav9 packaging a c5 12 microdystrophin flag expression cassette in non human primates |
| topic | adeno-associated virus promoter systemic delivery non-human primate dystrophin AAV |
| url | http://www.sciencedirect.com/science/article/pii/S2329050125000063 |
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