LGMD2A

Development of a strategy to treat limb-girdle muscular dystrophy (LGMD2A) using combined cell and gene therapy strategies

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 Obiettivo del progetto (Objective)

'In Duchenne muscular dystrophy, several mutations can be corrected by the exon-skipping technique on cells. In limb-girdle muscular dystrophy type 2A (LGMD2A), however, all the mutations described to date do not seem to be good candidates for the RNA correction technology. The recently identified c.17821072 G>C mutation in LGMD2A patients could be potentially repaired at RNA level. The principal aim of this project is to mask the effect of this mutation in stem cells from patients that can be derived to muscle and to test its therapeutic potential in a mouse model. This is the first step to be considered before planning a clinical trial in patients with this mutation. Our long term goal is to generate new advanced therapy protocols (gene and cell therapies) against LGMD2A, a muscular dystrophy that is specially prevailing in the population of the Basque region. To this end, we will pursue the following aims: 1. To develop a therapeutic approach for LGMD2A patients who carry c.17821072G>C mutation. 2. Isolation, optimization of the in vitro culture and genetic correction of CD133 cells from patients with this mutation. 3. Analysis of the efficacy of this combined therapeutic approach in murine models in vivo. In order to accomplish aims 1 and 2, we will develop a descriptive study with fibroblasts (from skin) and CD133 cells (from blood and muscle) from donors, in which we will determine the efficiency, yield and myogenic potential for subsequent testing in mice. To accomplish aim 3 we will perform a double blind experimental approach in mice (wild type versus C3KO/scid and mdx/scid mice), using CD133 cells with the most myogenic potential (from aim 2) as an in vivo therapy to determine the correction effect in the human calpain 3. This work would contribute to the treatment of LGMD2A patients who carry c.17821072G>C mutation in calpain 3 and it could also be helpful for the treatment of other patients with similar type of mutations.'

Introduzione (Teaser)

EU research is tackling a type of muscular dystrophy (MD) common in the Basque region for which there is no cure so far.

Descrizione progetto (Article)

MD is characterised by the production of defective muscle resulting in progressive skeletal muscle weakness and death of muscle cells and tissue. There are several forms of MD. Several mutations in the Calpain 3 gene cause limb-girdle muscular dystrophy type 2A (LGMD2A), for example, and unfortunately, no treatment currently exists for this type of muscular dystrophy, which is common in the Basque region. Testing in a cellular model, the EU-funded LGMD2A project aimed to remedy this situation.

To test the therapeutic approaches ex-vivo, the researchers developed a novel functional human cellular model of LGMD2A that shows spontaneous contractile capacity. The result is human myotubes that show a higher maturation, more comparable to muscle fibres.

The human myotubes differentiated under extracellular matrix scaffolds are a very promising test model, since they exhibit spontaneous calcium waves preceding contractility and a well-developed sarcomeric pattern. Human myotubes with Calpain 3 deficiency show increased oxidative stress, increase resting intracellular calcium levels, and reduced expression of the main sarcoplasmic reticulum calcium pumps: SERCA1 and SERCA2. Addition of pharmacological compounds aiming to stabilise calcium levels and/or reduce oxidative stress is still being tested. Preliminary results look favourable and several compounds are able to decrease oxidative stress levels of dystrophic myotubes.

After completion of all tests, the researchers intend to patent the most promising drug with clinical potential. LGMD2A scientists expect that the model system will assist in the development of a combined therapy for the disease and will expand the knowledge base. Applications can also extend to treatments for other muscular and cardiac diseases.