A5048165852- Muscle Physiology and Disorders
- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- Viral Infectious Diseases and Gene Expression in Insects
- RNA Interference and Gene Delivery
- Mitochondrial Function and Pathology
- Genetic Neurodegenerative Diseases
- RNA Research and Splicing
- Genetics and Neurodevelopmental Disorders
- Neuroscience and Neuropharmacology Research
- Silk-based biomaterials and applications
- Nerve injury and regeneration
- Animal Genetics and Reproduction
- Parkinson's Disease Mechanisms and Treatments
- Collagen: Extraction and Characterization
- Bone and Dental Protein Studies
- Biosimilars and Bioanalytical Methods
BioCruces Health research Institute
2019-2025
Utrophin is an autosomal paralogue of dystrophin, a protein whose deficit causes Duchenne and Becker muscular dystrophies (DMD/BMD). naturally overexpressed at the sarcolemma mature dystrophin-deficient fibres in DMD BMD patients as well mdx mouse model. Dystrophin utrophin can co-localise human foetal muscle, dystrophin-competent from DMD/BMD carriers, revertant fibre clusters biopsies patients. These findings suggest that overexpression could act surrogate, compensating for lack and, such,...
Myotonic dystrophy type I (DM1) is caused by CTG repeat expansions in the DMPK gene leading to mRNA toxicity and sequestration of splicing regulator MBNL1, affecting many tissues. We have developed an vitro screening platform based on ddPCR in-cell western quantify these mRNAs proteins characterised more than 20 cell models define DM1 biomarkers that could be useful for drug screening. protein levels were reduced DM1-immortalised myoblasts myotubes, but not fibroblasts, while MBNL1 was...
Gene editing methods are an attractive therapeutic option for Duchenne muscular dystrophy, and they have immediate application in the generation of research models. To generate myoblast cultures that could be useful vitro drug screening, we optimised a CRISPR/Cas9 gene edition protocol. We successfully used it wild type immortalised myoblasts to delete exon 52 dystrophin gene, modelling common dystrophy mutation; patient's deleted inhibitory microRNA target region utrophin UTR, leading...
Abstract Several exon skipping antisense oligonucleotides (eteplirsen, golodirsen, viltolarsen, and casimersen) have been approved for the treatment of Duchenne muscular dystrophy, but many more are in development targeting an array different DMD exons. Preclinical screening new oligonucleotide sequences is routinely performed using patient-derived cell cultures, evaluation their efficacy may be at RNA and/or protein level. While several methods to assess dystrophin expression culture...
Abstract Aims This study aims to develop a quantitative method for assessing collagen VI expression in cell cultures, which is crucial the diagnosis and treatment of VI-related dystrophies. Methods We developed combined in-cell western (ICW) on-cell (OCW) assay, that we have called ‘collablot’ quantify its organisation extracellular matrix cultures from patients healthy controls. To optimise it, optimised density protocols induce as well fixation permeabilisation methods. was completed with...
ABSTRACT Gene editing therapies in development for correcting out-of-frame DMD mutations Duchenne muscular dystrophy aim to replicate benign spontaneous deletions. Deletion of 45 – 55 exons (del45 55) was described asymptomatic subjects, but recently serious skeletal and cardiac complications have been reported. Uncovering why a single mutation like del45–55 is able induce diverse phenotypes grades severity may impact the strategies emerging therapies. Cellular models are essential this...
Abstract Gene edition methods are an attractive putative therapeutic option for Duchenne muscular dystrophy and they have immediate application in the generation of research models. To generate two new edited myoblast cultures that could be useful vitro drug screening, we optimised a CRISPR/Cas9 gene protocol. We successfully used it wild type immortalised myoblasts to delete exon 52 dystrophin gene: DMDΔ52-Model, modelling common mutation; patient’s deleted inhibitory microRNA target region...