Augmentation de l'expression de la chaine α1 de la laminine 111, un potentiel traitement pour la Dystrophie musculaire de Duchenne
|Abstract:||Laminin-111 protein complex links the extra-cellular matrix to integrin α7β1 in sarcolemma, thus replacing in dystrophic muscles links normally insured by dystrophin complex. Laminin-111 injection in mdx mouse increased expression of integrin α7β1, stabilized sarcolemma, restored serum creatine kinase to wild-type levels, and protected muscles from exercised-induced damages. These results suggested that increased Laminin-111 is a potential therapy for DMD. Laminin β1 and γ1 chains are expressed in adult human muscle but laminin α1 (LAMA1) gene is expressed only during embryogenesis. We thus developed an alternative method to Laminin-111 protein repeated administration by inducing expression of the endogenous LAMA1 gene. This was done with the CRSPR/Cas9 system, i.e., by targeting the LAMA1 promoter with one or several gRNAs and a dCas9 coupled with the VP160 transcription activation domain. LAMA1 mRNA (qRT-PCR) and proteins (immunohistochemistry and western blot) were not detected in the control C2C12 myoblasts. However, significant expression was observed in cells transfected and in mouse muscles electroporated with plasmids coding for dCas9-VP160 a gRNA. Larger synergic increases were observed by using 2 or 3 gRNAs. Increased expression of LAMA1 by the CRISPR/Cas9 system will have to be further investigated to verify whether this could be a treatment for several myopathies.|
|Document Type:||Mémoire de maîtrise|
|Open Access Date:||24 April 2018|
|Collection:||Thèses et mémoires|
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