A5088050328- Skin and Cellular Biology Research
- Silk-based biomaterials and applications
- Hearing, Cochlea, Tinnitus, Genetics
- RNA Interference and Gene Delivery
- Immune Cell Function and Interaction
- Cell Adhesion Molecules Research
- CRISPR and Genetic Engineering
- RNA regulation and disease
- Autoimmune Bullous Skin Diseases
- T-cell and B-cell Immunology
- melanin and skin pigmentation
- Cellular Mechanics and Interactions
- Vestibular and auditory disorders
- Wnt/β-catenin signaling in development and cancer
- Immunodeficiency and Autoimmune Disorders
- Animal Genetics and Reproduction
- Transgenic Plants and Applications
- Skin Protection and Aging
- Viral-associated cancers and disorders
- 3D Printing in Biomedical Research
- Tissue Engineering and Regenerative Medicine
- Cancer-related molecular mechanisms research
- Systemic Lupus Erythematosus Research
- Antimicrobial Peptides and Activities
- Congenital heart defects research
Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas
2016-2024
Hospital Universitario Fundación Jiménez Díaz
2016-2024
Centre for Biomedical Network Research on Rare Diseases
2009-2024
Centro de Investigación Biomédica en Red
2017-2024
Universidad Carlos III de Madrid
2013-2024
Unidades Centrales Científico-Técnicas
2019
Instituto de Investigación de Enfermedades Raras
2017
Instituto Cajal
2011
Instituto Ramón y Cajal de Investigación Sanitaria
2011
Hospital Universitario Ramón y Cajal
2003-2009
Gene editing constitutes a novel approach for precisely correcting disease-causing gene mutations. Frameshift mutations in COL7A1 causing recessive dystrophic epidermolysis bullosa are amenable to open reading frame restoration by non-homologous end joining repair-based approaches. Efficient targeted deletion of faulty exons polyclonal patient keratinocytes would enable the translation this therapeutic strategy clinic. In study, using dual single-guide RNA (sgRNA)-guided Cas9 nuclease...
Abstract Recent advances in molecular biology have led to the CRISPR revolution, but lack of an efficient and safe delivery system into cells tissues continues hinder clinical translation approaches. Polymeric vectors offer attractive alternative viruses as due their large packaging capacity safety profile. In this paper, we demonstrated potential use a highly branched poly( β -amino ester) polymer, HPAE-EB, enable genomic editing via CRISPRCas9-targeted excision exon 80 COL7A1 gene, through...
The prevalence of DFNA8/DFNA12 (DFNA8/12), a type autosomal dominant nonsyndromic hearing loss (ADNSHL), is unknown as comprehensive population-based genetic screening has not been conducted. We therefore completed unbiased for TECTA mutations in Spanish cohort 372 probands from ADNSHL families. Three additional families (Spanish, Belgian, and English) known to be linked DFNA8/12 were also included the screening. In an 835 American families, we preselected 73 based on audiometric data....
Clonal gene therapy protocols based on the precise manipulation of epidermal stem cells require highly efficient gene-editing molecular tools. We have combined adeno-associated virus (AAV)-mediated delivery donor template DNA with transcription activator-like nucleases (TALE) expressed by adenoviral vectors to address correction c.6527insC mutation in COL7A1 gene, causing recessive dystrophic epidermolysis bullosa a high percentage Spanish patients. After transduction these viral vectors,...
Recessive dystrophic epidermolysis bullosa (RDEB), Kindler syndrome (KS) and xeroderma pigmentosum complementation group C (XPC) are three cancer‐prone genodermatoses whose causal genetic mutations cannot fully explain, on their own, the array of associated phenotypic manifestations. Recent evidence highlights role stromal microenvironment in pathology these disorders. To investigate, by means comparative gene expression analysis, played dermal fibroblasts pathogenesis RDEB, KS XPC. We...
Abstract The biological role in vivo of the homologous CD3γ and δ invariant chains within human TCR/CD3 complex is a matter debate, as murine models do not recapitulate immunodeficiencies. We have characterized, Turkish family, two new patients with complete deficiency SCID symptoms compared them three CD3γ-deficient individuals belonging to families from Turkey Spain. All tested shared similar immunological features such partial expression defect, mild αβ γδ T lymphocytopenia, poor vitro...
Here we report the functional assessment of two novel deafness-associated γ-actin mutants, K118N and E241K, in a spectrum different situations with increasing biological complexity by combining biochemical cell analysis yeast mammalian cells. Our vivo experiments showed that while had very mild effect on behaviour, phenotype caused E241K mutation was severe characterized highly compromised ability to grow glycerol as carbon source, an aberrant multi-vacuolar pattern deposition thick F-actin...
Recessive dystrophic epidermolysis bullosa is a severe skin fragility disease caused by loss of functional type VII collagen at the dermal-epidermal junction. A frameshift mutation in exon 80 COL7A1 gene, c.6527insC, highly prevalent Spanish patient population. We have implemented gene-editing strategies for frame restoration NHEJ-induced indels epidermal stem cells from patients carrying this mutation. TALEN nucleases designed to cut within sequence were delivered primary keratinocyte...
Genome-editing technologies that enable the introduction of precise changes in DNA sequences have potential to lead a new class treatments for genetic diseases. Epidermolysis bullosa (EB) is group rare disorders characterized by extreme skin fragility. The recessive dystrophic subtype EB (RDEB), which has one most severe phenotypes, caused mutations COL7A1. In this study, we report gene-editing approach ex vivo homology-directed repair (HDR)-based gene correction uses CRISPR-Cas9 system...
T cells recognize antigens via their cell surface TCR and are classified as either αβ or γδ depending on the variable chains in TCR, α β γ δ, respectively. Both TCRs also contain several invariant chains, including CD3δ, which support expression transduce signal. Mutations would be expected to affect a single lineage, while mutations all cells. Consistent with this, CD3δ-deficient patients described date showed complete block development. However, CD3δ-KO mice have an cell-specific defect....
Recessive dystrophic epidermolysis bullosa, a devastating skin fragility disease characterized by recurrent blistering, scarring, and high risk of developing squamous cell carcinoma is caused mutations in COL7A1, the gene encoding type VII collagen, which major component anchoring fibrils that bind dermis epidermis. Ex vivo correction COL7A1 editing patients' cells has been achieved before. However, approaches are necessary to address direct treatment blistering lesions characteristic this...
Current efforts to find specific genodermatoses treatments and define precise pathogenesis mechanisms require appropriate surrogate models with human cells. Although transgenic gene knockout mouse for several of these disorders exist, they often fail faithfully replicate the clinical histopathological features skin condition. We have established a highly efficient method deletion critical sequences in primary keratinocytes, based on CRISPR-Cas9-mediated editing. Using this methodology,...
Abstract Epidermolysis bullosa with pyloric atresia ( EB ‐ PA ) is a rare autosomal recessive hereditary disease variable prognosis from lethal to very mild. classified into Simplex form EBS : OMIM #612138) and Junctional JEB #226730), it caused by mutations in ITGA 6 , ITGB 4 PLEC genes. We report the analysis of six patients including two dizygotic twins. Skin immunofluorescence epitope mapping was performed followed PCR direct sequencing gene. Two presented non‐lethal associated missense...
Functional analysis in mouse models is necessary to establish the involvement of a set genetic variations tumor development. A modeling platform facilitate and cost-effectively analyze role multiple genes carcinogenesis would be valuable. Here, we present an innovative strategy for lung mutagenesis using CRISPR/Cas9 ribonucleoproteins delivered via cationic polymers. This approach allows simultaneous inactivation genes. We validate effectiveness this system by targeting group suppressor...
Abstract Recessive dystrophic epidermolysis bullosa (RDEB) is a rare genetic disease caused by loss of function mutations in the gene coding for collagen VII (C7) due to deficient or absent C7 expression. This disrupts structural and functional skin architecture, leading blistering, chronic wounds, inflammation, important systemic symptoms affecting mouth, gastrointestinal tract, cornea, kidney function, an increased cancer risk. RDEB patients have extremely poor quality life often die at...
Abstract Dystrophic epidermolysis bullosa is a rare blistering condition caused by mutations in the COL 7A1 gene. Different clinical variants have been described, with dominant and recessive inheritance, but no consistent findings elucidated to establish genotype–phenotype correlation. We present three unrelated patients two identical pathogenic compound heterozygous gene that developed different forms of dystrophic bullosa—epidermolysis pruriginosa mild non‐Hallopeau–Siemens—raising...
AbstractBackground: T lymphocytes play a crucial role in the pathogenesis of inflammatory bowel disease. Achieving stable T-cell lines, rather than continuous bleeding patients, is desirable order to dissect their implication Methods: Long-lasting lines from patients with Crohn disease and ulcerative colitis healthy volunteers have been obtained by transformation using lymphotropic Herpesvirus saimiri. Lines were subjected phenotypic functional analyses, results compared freshly isolated...
To dissect the phenotypic and functional features of mucosal T lymphocytes in patients with gastric adenocarcinoma, we have used Herpesvirus saimiri transformation procedure to achieve T‐cell lines from origin. Once achieved, cell function was assessed by vitro stimulation mitogens. CD2‐specific monoclonal antibodies (α‐CD2), alone or combination interleukin (IL)‐2, rendered fewer counts (34 408±3965 52 157±6473 c.p.m., respectively) than controls (67 471±11 755 P <0.01 77 864±12 545...