C9orf72 repeat expansion-tuned allelic suppression by CRISPRi as an ALS therapy
Grant Award Details
Grant Type:
Grant Number:
DISC2-15010
Investigator(s):
Disease Focus:
Human Stem Cell Use:
Award Value:
$2,274,768
Status:
Active
Grant Application Details
Application Title:
C9orf72 repeat expansion-tuned allelic suppression by CRISPRi as an ALS therapy
Public Abstract:
Research Objective
We aim to discover an adeno-associated viral (AAV) CRISPRi gene therapy for amyotrophic lateral sclerosis (ALS) patients with hexanucleotide repeat expansions in the C9orf72 gene.
Impact
This therapy can be administered intravenously once, have long lasting effects, and is indicated for all ALS patients who carry repeat expansions of varying lengths and toxicity in the C9orf72 gene.
Major Proposed Activities
We aim to discover an adeno-associated viral (AAV) CRISPRi gene therapy for amyotrophic lateral sclerosis (ALS) patients with hexanucleotide repeat expansions in the C9orf72 gene.
Impact
This therapy can be administered intravenously once, have long lasting effects, and is indicated for all ALS patients who carry repeat expansions of varying lengths and toxicity in the C9orf72 gene.
Major Proposed Activities
- Establish a high-throughput cellular platform to screen for an optimized small guide RNA (sgRNA) and CRISPRi system that selectively silences long, pathogenic C9orf72 repeat expansions.
- Generate neural cells from C9orf72 ALS patient induced pluripotent stem cells (iPSCs) and optimize the AAV CRISPRi system to reduce toxic RNA and protein products caused by C9orf72 repeat expansions.
- Test the reproducibility of the AAV CRISPRi system to reduce toxic RNA and protein products in iPSC models of 15 female and 15 male C9orf72 ALS patients while assessing any off-target genomic effects.
- Optimize the AAV CRISPRi therapy in mice genetically modified with pathogenic C9orf72 repeat expansions and test its ability to reduce toxic RNA and protein products while preserving motor function.
Statement of Benefit to California:
ALS currently affects thousands of Californians with no cure. Patients rely on full-time caregivers, and underserved communities have a disproportionate burden. We use a rational combination of technologies to treat this genetic form of ALS caused by DNA repeat expansions. If we are successful, our approach can readily transfer to other repeat expansion diseases in neurodegeneration. This would markedly alleviate the neurodegenerative disease risks facing a rapidly aging California population.