Spinal subpial delivery of AAV9 enables widespread gene silencing and blocks motoneuron degeneration in ALS

• Published in: Nature medicine Vol. 26; no. 1; pp. 118 - 130
• Main Authors: Bravo-Hernandez, Mariana, Tadokoro, Takahiro, Navarro, Michael R., Platoshyn, Oleksandr, Kobayashi, Yoshiomi, Marsala, Silvia, Miyanohara, Atsushi, Juhas, Stefan, Juhasova, Jana, Skalnikova, Helena, Tomori, Zoltan, Vanicky, Ivo, Studenovska, Hana, Proks, Vladimir, Chen, PeiXi, Govea-Perez, Noe, Ditsworth, Dara, Ciacci, Joseph D., Gao, Shang, Zhu, Wenlian, Ahrens, Eric T., Driscoll, Shawn P., Glenn, Thomas D., McAlonis-Downes, Melissa, Da Cruz, Sandrine, Pfaff, Samuel L., Kaspar, Brian K., Cleveland, Don W., Marsala, Martin
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.01.2020; Nature Publishing Group
• Subjects: 692/617/375/1917/1285; 692/617/375/364; Amyotrophic lateral sclerosis; Amyotrophic Lateral Sclerosis - genetics; Amyotrophic Lateral Sclerosis - physiopathology; Amyotrophic Lateral Sclerosis - therapy; Animals; Atrophy; Biomedical and Life Sciences; Biomedicine; Brain; Cancer Research; Care and treatment; Degeneration; Dependovirus - metabolism; Disease Progression; Evoked Potentials, Motor; Female; Gene Expression Regulation; Gene Silencing; Gene therapy; Gene transfer; Gene Transfer Techniques; Genetic aspects; Genetic vectors; Health aspects; Humans; Infectious Diseases; Inflammation - pathology; Injection; Innervation; Interneurons - pathology; Male; Metabolic Diseases; Methods; Mice, Inbred C57BL; Mice, Transgenic; Molecular Medicine; Motor neurons; Motor Neurons - pathology; Muscle Development; Muscles; Mutants; Nerve Degeneration - genetics; Nerve Degeneration - physiopathology; Nerve Degeneration - therapy; Neurodegeneration; Neurodegenerative diseases; Neurosciences; Pia Mater - pathology; Pia Mater - physiopathology; Preservation; Primates; Protein Folding; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA, Small Interfering - administration & dosage; Spinal cord; Spinal Cord - diagnostic imaging; Spinal Cord - pathology; Spinal Cord - physiopathology; Substantia grisea; Superoxide dismutase; Superoxide Dismutase-1 - genetics; Superoxide Dismutase-1 - metabolism; Swine; Thorax; Viruses; United States
• ISSN: 1078-8956; 1546-170X; 1546-170X
• DOI: 10.1038/s41591-019-0674-1

Gene silencing with virally delivered shRNA represents a promising approach for treatment of inherited neurodegenerative disorders. In the present study we develop a subpial technique, which we show in adult animals successfully delivers adeno-associated virus (AAV) throughout the cervical, thoracic and lumbar spinal cord, as well as brain motor centers. One-time injection at cervical and lumbar levels just before disease onset in mice expressing a familial amyotrophic lateral sclerosis (ALS)-causing mutant SOD1 produces long-term suppression of motoneuron disease, including near-complete preservation of spinal α-motoneurons and muscle innervation. Treatment after disease onset potently blocks progression of disease and further α-motoneuron degeneration. A single subpial AAV9 injection in adult pigs or non-human primates using a newly designed device produces homogeneous delivery throughout the cervical spinal cord white and gray matter and brain motor centers. Thus, spinal subpial delivery in adult animals is highly effective for AAV-mediated gene delivery throughout the spinal cord and supraspinal motor centers. Injection of AAV–shRNA below the pial surface of the spinal cord prevents onset or ameliorates progression in a mouse model of ALS, and achieves widespread delivery to the spinal cord and brain motor centers in adult pigs and non-human primates.