In vivo genome editing using novel AAV-PHP variants rescues motor function deficits and extends survival in a SOD1-ALS mouse model

• Published in: Gene therapy Vol. 30; no. 5; pp. 443 - 454
• Main Authors: Chen, Yi A., Kankel, Mark W., Hana, Sam, Lau, Shukkwan Kelly, Zavodszky, Maria I., McKissick, Olivia, Mastrangelo, Nicole, Dion, Jessica, Wang, Bin, Ferretti, Daniel, Koske, David, Lehman, Sydney, Koszka, Kathryn, McLaughlin, Helen, Liu, Mei, Marshall, Eric, Fabian, Attila J., Cullen, Patrick, Marsh, Galina, Hamann, Stefan, Craft, Michael, Sebalusky, Jennifer, Arnold, H. Moore, Driscoll, Rachelle, Sheehy, Adam, Luo, Yi, Manca, Sonia, Carlile, Thomas, Sun, Chao, Sigrist, Kirsten, McCampbell, Alexander, Henderson, Christopher E., Lo, Shih-Ching
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-05-2023; Nature Publishing Group
• Subjects: 42/41; 42/44; 631/61/201; 631/61/2300; 64/110; 64/60; 82/1; Action potential; Amyotrophic lateral sclerosis; Amyotrophic Lateral Sclerosis - genetics; Amyotrophic Lateral Sclerosis - therapy; Animals; Atrophy; Biomedical and Life Sciences; Biomedicine; Cell Biology; CRISPR; Denervation; Disease Models, Animal; Gene Editing; Gene Expression; Gene Therapy; Genome editing; Genomes; Hereditary diseases; Human Genetics; Humans; Life span; Mice; Mice, Transgenic; Nanotechnology; Neonates; Neuromuscular junctions; Spinal cord; Superoxide dismutase; Superoxide Dismutase - genetics; Superoxide Dismutase - metabolism; Superoxide Dismutase-1 - genetics; Therapeutic targets
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/s41434-022-00375-w

CRISPR-based gene editing technology represents a promising approach to deliver therapies for inherited disorders, including amyotrophic lateral sclerosis (ALS). Toxic gain-of-function superoxide dismutase 1 (SOD1) mutations are responsible for ~20% of familial ALS cases. Thus, current clinical strategies to treat SOD1-ALS are designed to lower SOD1 levels. Here, we utilized AAV-PHP.B variants to deliver CRISPR-Cas9 guide RNAs designed to disrupt the human SOD1 (huSOD1) transgene in SOD1
G93A
mice. A one-time intracerebroventricular injection of AAV.PHP.B-huSOD1-sgRNA into neonatal H11
Cas9
SOD1
G93A
mice caused robust and sustained mutant huSOD1 protein reduction in the cortex and spinal cord, and restored motor function. Neonatal treatment also reduced spinal motor neuron loss, denervation at neuromuscular junction (NMJ) and muscle atrophy, diminished axonal damage and preserved compound muscle action potential throughout the lifespan of treated mice. SOD1
G93A
treated mice achieved significant disease-free survival, extending lifespan by more than 110 days. Importantly, a one-time intrathecal or intravenous injection of AAV.PHP.eB-huSOD1-sgRNA in adult H11
Cas9
SOD1
G93A
mice, immediately before symptom onset, also extended lifespan by at least 170 days. We observed substantial protection against disease progression, demonstrating the utility of our CRISPR editing preclinical approach for target evaluation. Our approach uncovered key parameters (e.g., AAV capsid, Cas9 expression) that resulted in improved efficacy compared to similar approaches and can also serve to accelerate drug target validation.