Objectives Primary objectives of our project are i) to characterise myogenic cells from 14 DMD boys with dystrophin deletions capable of being modulated by means of exon 46 and 51 skipping, in order to make them eligible for the approaching European multicentric trial; ii) approval from the Istituto Superiore di Sanità of the GMP molecules which will be used in the multicentric trials. Secondary objective is the evaluation of dystrophin transcriptome changes in patients’ cells. Background/Rationale Antisense oligoribonucleotides are able to induce exon skipping in the dystrophin hnRNA therefore by−passing out−of−frame deletions. The rationale of this approach is based on the fact that the majority of DMDs is due to large deletions causing the disruption of the frame. The AONs exon skipping generates internally deleted, but functional dystrophin proteins and would convert a severe DMD into a milder phenotype. A successful first−in−man trial has recently been completed on DMD and a multicentric trial is approaching. Description of the project We aim at performing on patient’s cells transfected with AONs the following studies: a) Real−Time RT−PCR in order to finely detect and quantify non−skipped as well as skipped transcripts; b) functional effect of the AONs treatment by evaluating dystrophin protein rescue; c) effect of AONs modulation on the cell transcriptome by using an array−based strategy able to both define the dystrophin transcription atlas and verify the correct dystrophin full mRNA composition after the treatment. Genomic studies will be carried for mutation confirmation and SNPs occurrence. Anticipated output These studies will fully characterise AONs−treated cells of the selected Italian DMD patients making them eligible and facilitating their inclusion in the first multicentric AONs trial for dystrophinopathies. The ISS audit will allow us to obtain permission and ethical approval for the trial.

Antisense modulation and characterisation of myogenic cells from 14 boys with Duchenne muscular dystrophy as pre-trial study aimed at patients’ recruitment for an European multicentric clinical trial (Telethon UILDM-GGPO7011)

FERLINI, Alessandra
2008

Abstract

Objectives Primary objectives of our project are i) to characterise myogenic cells from 14 DMD boys with dystrophin deletions capable of being modulated by means of exon 46 and 51 skipping, in order to make them eligible for the approaching European multicentric trial; ii) approval from the Istituto Superiore di Sanità of the GMP molecules which will be used in the multicentric trials. Secondary objective is the evaluation of dystrophin transcriptome changes in patients’ cells. Background/Rationale Antisense oligoribonucleotides are able to induce exon skipping in the dystrophin hnRNA therefore by−passing out−of−frame deletions. The rationale of this approach is based on the fact that the majority of DMDs is due to large deletions causing the disruption of the frame. The AONs exon skipping generates internally deleted, but functional dystrophin proteins and would convert a severe DMD into a milder phenotype. A successful first−in−man trial has recently been completed on DMD and a multicentric trial is approaching. Description of the project We aim at performing on patient’s cells transfected with AONs the following studies: a) Real−Time RT−PCR in order to finely detect and quantify non−skipped as well as skipped transcripts; b) functional effect of the AONs treatment by evaluating dystrophin protein rescue; c) effect of AONs modulation on the cell transcriptome by using an array−based strategy able to both define the dystrophin transcription atlas and verify the correct dystrophin full mRNA composition after the treatment. Genomic studies will be carried for mutation confirmation and SNPs occurrence. Anticipated output These studies will fully characterise AONs−treated cells of the selected Italian DMD patients making them eligible and facilitating their inclusion in the first multicentric AONs trial for dystrophinopathies. The ISS audit will allow us to obtain permission and ethical approval for the trial.
Ferlini, Alessandra
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11392/1679300
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