Mutations in CLN7 cause the lysosomal storage, dysfunction, and neurodegeneration in Batten’s disease, however the mechanisms that lead this dysfunction are not known. Our previous studies in patient fibroblasts indicated CLN7 mutations cause autophagic dysfunction and reduction of lysosomal hydrolase activity. Since CLN7 results in prominent neurodegeneration, we propose to develop novel neuronal models of Batten’s disease through reprogramming patient fibroblasts into induced pluripotent stem cells (iPSCs). We will employ robust and highly reproducible protocols to differentiate iPSC’s into cortical type neurons to provide a model that is highly relevant to Batten’s disease. Using CRISPR/Cas9 gene correction, we will generate isogenic controls to be used during our phenotype analysis. Induced cortical neurons will be subjected to our established biochemical and imaging assays to probe lysosomal function, autophagic flux, and storage material by ultrastructural analysis. Finally, we will determine the efficiency of investigational therapeutics aimed at restoring CLN7 expression, including oligonucleotides that improve the stability of CLN7 mutant transcripts, and adeno-associated viral expression of wild-type CLN7. At the end of the funding period, we hope to have provided a novel, accurate model of CLN7 Batten’s disease that recapitulates several features of the disease. This model can be employed to probe disease mechanism, test novel therapies, and be distributed to other researchers with expertise in other fields.
|Effective start/end date||6/1/19 → 5/31/21|
- Mila's Miracle Foundation, Inc. (AGMT 5/7/2019)