A5084705827- Cystic Fibrosis Research Advances
- Neonatal Respiratory Health Research
- Tracheal and airway disorders
- Inhalation and Respiratory Drug Delivery
- Advanced biosensing and bioanalysis techniques
- CRISPR and Genetic Engineering
- Delphi Technique in Research
- Genomics and Rare Diseases
- Microfluidic and Bio-sensing Technologies
- Energy Harvesting in Wireless Networks
- Pharmaceutical studies and practices
- 3D Printing in Biomedical Research
- Asthma and respiratory diseases
- Neurogenetic and Muscular Disorders Research
- RNA and protein synthesis mechanisms
- Gut microbiota and health
- RNA Interference and Gene Delivery
- Child Nutrition and Feeding Issues
- Congenital Ear and Nasal Anomalies
- Inflammatory Bowel Disease
- Cancer Cells and Metastasis
- Animal Genetics and Reproduction
- Biomedical and Engineering Education
- Digital Holography and Microscopy
- Image Processing Techniques and Applications
KU Leuven
2016-2025
VIB-KU Leuven Center for Microbiology
2020-2021
Wilhelmina Children's Hospital
2020
Organ Recovery Systems (Belgium)
2019
UCLouvain
2017
Newcastle University
2017
Hadassah Medical Center
2017
University of Lisbon
2002-2015
National Institute of Health Dr. Ricardo Jorge
2003-2013
Inserm
2013
Prime editing is a recent, CRISPR-derived genome technology capable of introducing precise nucleotide substitutions, insertions, and deletions. Here, we present prime approaches to correct L227R- N1303K-CFTR, two mutations that cause cystic fibrosis are not eligible for current market-approved modulator therapies. We show that, upon DNA correction the CFTR gene, complex glycosylation, localization, and, most importantly, function protein restored in HEK293T 16HBE cell lines. These findings...
This protocol describes the isolation, handling, culture of, and experiments with human colon stem cell organoids in context of cystic fibrosis (CF). In organoids, function transmembrane conductance regulator (CFTR) protein its rescue by CFTR modulators can be quantified using forskolin-induced swelling assay. Implementation procedures validation are described for six CF organoid lines, representative genotypes tested basal response to CFTR-modulating drugs. For complete details on use...
Given the vast number of cystic fibrosis transmembrane conductance regulator (CFTR) mutations, biomarkers predicting benefit from CFTR modulator therapies are needed for subjects with (CF).To study function in organoids common and rare mutations evaluate correlations between clinical data.Intestinal were grown rectal biopsies a cohort 97 CF. Residual was measured by quantifying organoid swelling induced forskolin response to modulators correctors, potentiator their combination. Organoid data...
Estimates of the level transcripts from cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene required to develop a CF phenotype range 4-20% normal. Due importance obtaining reliable data on this issue for therapeutic strategies, we developed novel polymerase chain reaction-based method quantify CFTR and applied it analysis nasal epithelium RNA five patients with 3272-26A>G/F508del genotype. We calculated that 8.2 +/- 0.84% total present in these is normal full-length mRNA....
Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the CFTR gene. The 3272-26A>G and 3849+10kbC>T alter correct splicing of gene, generating new acceptor donor splice sites respectively. Here we develop a genome editing approach to permanently these genetic defects, using single crRNA Acidaminococcus sp. BV3L6, AsCas12a. This repair strategy highly precise, showing very strong discrimination between wild-type mutant sequence complete absence detectable off-targets....
Abstract Background and Aims In vitro studies using immortalised cancer cell lines showed that butyrate has an overall positive effect on epithelial barrier integrity, but the physiological relevance of is limited. We developed monolayers from human tissue samples patients with ulcerative colitis [UC] to assess function. Methods A protocol establish primary cells UC [n = 10] non-UC controls was optimised. The were treated 8 mM sodium ± tumour necrosis factor alpha [TNFα] type II interferon...
Cystic fibrosis (CF) is a genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. The 2789+5G>A CFTR mutation quite frequent defect causing an aberrant splicing and non-functional protein. Here we used CRISPR adenine base editing (ABE) approach to correct absence of DNA double-strand breaks (DSB). To select strategy, developed minigene cellular model reproducing defect. We obtained up 70% adapting ABE PAM sequence optimal for targeting with SpCas9-NG...
Cystic Fibrosis (CF) is caused by ∼1,900 mutations in the CF transmembrane conductance regulator (CFTR) gene encoding for a cAMP-regulated chloride (Cl(-)) channel expressed several epithelia. Clinical features are dominated respiratory symptoms, but there variable organ involvement thus causing diagnostic dilemmas, especially non-classic cases.To further establish measurement of CFTR function as sensitive and robust biomarker diagnosis prognosis CF, we herein assessed cholinergic...
Gene therapy holds promise for a curative mutation-independent treatment applicable to all patients with cystic fibrosis (CF). The various viral vector-based clinical trials conducted in the past have demonstrated safety and tolerance of different vectors, but none led clear persistent benefit. Recent breakthroughs recombinant adeno-associated vector (rAAV)-based gene encouraged us reexplore an rAAV approach CF.We evaluated preclinical potential CF restore chloride fluid secretion two...
Assessment of the functional consequences variants near splice sites is a major challenge in diagnostic laboratory. To address this issue, we created expression minigenes (EMGs) to determine RNA and protein products generated by site (n = 10) implicated cystic fibrosis (CF). Experimental results were compared with splicing predictions eight silico tools. EMGs containing full-length Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) coding sequence flanking intron sequences wild-type...
Introduction Cystic fibrosis (CF) is a severe monogenic disorder caused by mutations in the cystic transmembrane conductance regulator ( CFTR ) gene. Several types of modulators (correctors/potentiators) have been developed to overcome protein dysfunction associated with these mutations. modulator therapy now available for major CF-causing mutations; however, 10% people CF remain without causal treatments. By combining investigational and market-approved modulators, we aimed maximise...
Background. The most common CFTR mutation, F508del, presents with multiple cellular defects. However, the possible defects caused by many rarer mutations are not well studied. We investigated four rare E60K, G85E, E92K and A455E against well-characterized mutations, F508del G551D, their responses to corrector VX-809 and/or potentiator VX-770. Methods. Using complementary assays in HEK293T stable cell lines, we determined maturation Western blotting, trafficking flow cytometry using...
Diagnosing cystic fibrosis (CF) when sweat chloride is not in the CF range and less than 2 disease-causing CFTR mutations are found requires physiological assays, which always feasible or available. We developed a new assay based on morphological differences between rectal organoids from subjects with without CF. In 167 22 CF, two parameters derived semi-automated image analysis protocol (rectal organoid morphology analysis, ROMA) fully discriminated non-CF (p<0.001). ROMA, at all ages,...
Background/Aims: Mutations in the CFTR gene cause Cystic Fibrosis (CF) most common life-threatening autosomal recessive disease affecting Caucasians. We identified a mutation (c.120del23) abolishing normal translation initiation codon, which occurs two Portuguese CF patients. This study aims at functionally characterizing effect of this novel mutation. Methods: RNA and protein techniques were applied to both native tissues from patients recombinant cells expressing constructs determine...
Cystic fibrosis (CF) is a life-threatening disorder characterised by decreased pulmonary mucociliary and pathogen clearance, an exaggerated inflammatory response leading to progressive lung damage. CF caused bi-allelic pathogenic variants of the cystic transmembrane conductance regulator (CFTR) gene, which encodes chloride channel. CFTR expressed in endothelial cells (ECs) EC dysfunction has been reported patients, but role for this ion channel ECs regarding disease progression poorly...