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Genome and transcriptome engineering with CRISPR/Cas as a precision medicine for Charcot-Marie-Tooth type 2L.

Patients with Charcot-Marie-Tooth disease (CMT) have a hereditary motor and sensory neuropathy causing a length-dependent degeneration of their peripheral nerves. Most patients have a duplication on chromosome 17 resulting in a higher expression of the peripheral myelin protein 22 (PMP22). Finding ways to reduce PMP22 expression has led to the development of a treatment for this group of patients. However, a significant number of CMT patients are incurable due to rare mutations in more than 90 different genes, complicating the development of effective treatment strategies. In this TOP project we aim to make use of the cutting-edge CRISPR/Cas technology to selectively eliminate or even correct a mutant transcript. As proof-of-concept we will focus on a dominant missense mutation occurring in the small heat shock protein HSPB8 causing an axonal subtype of CMT neuropathy. Complete deletion of this gene is only associated with mild (subclinical) symptoms. This therefore provides a therapeutic window where specific elimination of the mutant allele may lead to amelioration of the phenotype. We will thus first establish a proof-of-principle by introducing CRISPR/Cas9 with adeno-associated virus (AAV) to selectively inactivate the mutant allele in vivo. Then, we will assess if we can obtain the same specificity with CRISPR/Cas13b which selectively degrades RNA molecules and may thus reduce the risk of introducing permanent off-target effects. Finally, we will assess the possibility to correct mutant transcripts back to wild type with disabled Cas13b coupled base editors. These in vivo studies in the Hspb8 knock-in mouse model will be complemented by motor neurons differentiated from HSPB8 patient derived induced pluripotent stem cells (iPSCs). With these cutting-edge genome and transcriptome engineering approaches we aim to explore the potential of this approach for patients with peripheral neuropathies who cannot be treated with current medicines.
Date:1 Jan 2020  →  Today
Disciplines:Nucleic acids, Genome structure and regulation, Neurological and neuromuscular diseases