B5 - Neurologic Diseases (excluding Ophthalmic and Auditory Diseases)
1590: AAV-Mediated Gene Augmentation Therapy for NEK1-ALS
Type: Poster Session
Poster Board Number: 1590
Presentation Details
Session Title: Friday Posters: Neurologic Diseases
Amyotrophic lateral sclerosis (ALS) is a terminal neurodegenerative disease that affects upper and lower motor neurons, leading to progressive muscle weakening and respiratory failure. Loss-of-function mutations in NIMA-related kinase 1 (NEK1) constitute a major genetic cause of ALS in both familial and sporadic ALS cases, accounting for 2 to 3% of all cases. To develop and test a NEK1 gene augmentation therapy, we created a mouse model with a nonsense mutation in the mouse NEK1 gene. This mutation resulted in a complete absence of NEK1 protein in homozygous mice. Preliminary data show that homozygous mice are noticeably smaller and have a significantly reduced bodyweight measured from three weeks onwards compared to heterozygous and WT littermates. Mice are currently 16 weeks old; any effect on lifespan has yet to be determined. We will further establish other NEK1 phenotypes, such as motor disturbances, DNA damage, cilia formation and nuclear envelope integrity. In NEK1 patient-derived fibroblasts and induced motor neurons, we observed DNA damage and diminished nuclear envelope integrity. To rescue any behavioral and molecular phenotypes in vitro and in vivo, we are designing AAV-mediated NEK1 augmentation strategies. Thus far, heterozygous neonatal NEK1 mice received intracerebroventricular injections of AAV-PHP.eb-hNEK1 driven by the MSCV promoter. This resulted in 11.6 and 1.8 vg/diploid genomes and significant expression of hNEK1 transcripts in the brain and spinal cord, respectively. We are exploring diverse NEK1 augmentation strategies, including different NEK1 transcript variants, AAV serotypes, promoters, and regulatory elements. Our objective is to establish effective therapeutic interventions for NEK1-related ALS, evaluating outcomes in both homozygous NEK1 mice and patient-derived cells.
Katharina E. Meijboom1, Desiree M. Baron1, Debra Cameron1, Erinn Ives1, Aparna Sreeram1, Ana Rita Batista2, Janice A. Dominov1, Cathleen Lutz3, Robert H. Brown1, John E. Landers1
1UMASS Chan Medical School, Worcester, MA,2UMass Chan Medical School, Worcester, MA,3The Jackson Laboratory, Bar Harbor, ME"
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