Gene editing in Myotonic Dystrophy type 1: assessment of efficiency, safety and therapeutic effect of CTG-repeat deletion in a mouse model of disease (DSB.AD007.200)

Area tematica

Scienze biomediche

Area progettuale

Struttura responsabile del progetto di ricerca

Istituto di Biochimica e Biologia Cellulare (IBBC)

Responsabile di progetto

GERMANA FALCONE
Telefono: 06 90091323
E-mail: germana.falcone@cnr.it

Abstract

Myotonic dystrophy type 1 (DM1) is a dominantly inherited, multisystemic disorder caused by expanded CTG repeats in the 3' untranslated region (3'UTR) of the DMPK gene. DM1 is the most common adult-onset muscular dystrophy characterized by progressive skeletal muscle weakness, myotonia, cardiac arrhythmia, smooth muscle dysfunction and neurological abnormalities. DMPK mutated transcript accumulates into nuclear foci that affect the localization and activities of RNA-binding proteins involved in splicing regulation. To date no effective therapy is yet available for DM1.
A direct genomic approach for treating an autosomal dominant gain-of-function disease such as DM1 is the removal of the mutated gene region. The multiplex capability of CRISPR/Cas9 gene editing allows the deletion of unwanted genomic sequences by eliciting two simultaneous double-strand breaks. Having already generated and tested highly specific and inducible CRISPR/Cas9 components in DM1 patient-derived cells, we plan to apply this gene editing strategy in DMSXL mice in vivo with the ultimate goal to obtain the permanent elimination of the genetic defect and the reversal of the diseased phenotype.

Obiettivi

In this proposal we plan to apply this gene therapy approach in vivo in transgenic mice carrying a mutated human DMPKgene, that exhibit a pathologic neuromuscular phenotype similar to that observed in human DM1 disease.The project goal will be pursued through the following steps:
1. Assessment of optimal dosage and timing conditions for CRISPR/Cas9 expression and DMPK gene editing in diseased animals.
2. Determination of gene editing efficiency and specificityin CRISPR/Cas9 treated animals and human myogenic cells.
3. Analysis of disease recovery following systemic or muscle-restricted deletion of CTG repeatsin diseased animals.
4. Analysis of transcriptomic changes in skeletal muscles derived from CRISPR/Cas9 treated animals.
5. Characterization of satellite cells derived from CRISPR/Cas9 treated animals.
We believe that a detailed understanding of the CRISPR/Cas9 gene editing strategy in vivo is a pre-requisite for its efficient and safe perspective application in humans as a potential gene therapy for DM1.

Data inizio attività

01/01/2020

Parole chiave

myotonic dystrophy, gene therapy, mouse model

Ultimo aggiornamento: 07/06/2025