A5045740482- Cystic Fibrosis Research Advances
- Neonatal Respiratory Health Research
- Ion channel regulation and function
- Advanced biosensing and bioanalysis techniques
- Respiratory viral infections research
- Neuroscience and Neuropharmacology Research
- Cardiac electrophysiology and arrhythmias
- Pharmaceutical studies and practices
- Legume Nitrogen Fixing Symbiosis
- Inhalation and Respiratory Drug Delivery
- Ion Channels and Receptors
- RNA and protein synthesis mechanisms
- Nematode management and characterization studies
- Ion Transport and Channel Regulation
- RNA Research and Splicing
- Cellular transport and secretion
- Tracheal and airway disorders
- RNA modifications and cancer
- Pneumocystis jirovecii pneumonia detection and treatment
- Neuroblastoma Research and Treatments
- Infant Nutrition and Health
- DNA and Nucleic Acid Chemistry
- S100 Proteins and Annexins
- Bacterial Genetics and Biotechnology
- Ubiquitin and proteasome pathways
Istituto Giannina Gaslini
2014-2024
Istituti di Ricovero e Cura a Carattere Scientifico
2018-2024
Tecnologie Avanzate (Italy)
2008-2011
Calcium-dependent chloride channels are required for normal electrolyte and fluid secretion, olfactory perception, neuronal smooth muscle excitability. The molecular identity of these membrane proteins is still unclear. Treatment bronchial epithelial cells with interleukin-4 (IL-4) causes increased calcium-dependent channel activity, presumably by regulating expression the corresponding genes. We performed a global gene analysis to identify that regulated IL-4. Transfection specific small...
The TMEM16A protein has a potential role as Ca(2+)-activated Cl(-) channel (CaCC) in airway epithelia where it may be important the homeostasis of surface fluid. We investigated function and expression primary human bronchial epithelial cells cell line (CFBE41o-). Under resting conditions, was relatively low. However, silencing with short-interfering RNAs caused marked inhibition CaCC activity, thus demonstrating that low is sufficient to support Ca(2+)-dependent transport. Following...
Cystic fibrosis (CF) is caused by mutations in the CFTR chloride channel. Deletion of phenylalanine 508 (F508del), most frequent CF mutation, impairs maturation and gating protein. Such defects may be corrected vitro pharmacological modulators named as correctors potentiators, respectively. We have evaluated a panel potentiators derived from various sources to assess potency, efficacy, mechanism action. For this purpose, we used functional biochemical assays on two different cell expression...
Deletion of phenylalanine at position 508 (F508del) in the CFTR chloride channel is most frequent mutation cystic fibrosis (CF) patients. F508del impairs stability and folding protein, thus resulting mistrafficking premature degradation. F508del-CFTR defects can be overcome with small molecules termed correctors. We investigated efficacy properties VX-445, a newly developed corrector, which one three active principles present drug (Trikafta®/Kaftrio®) recently approved for treatment CF...
SCN(-) (thiocyanate) is an important physiological anion involved in innate defense of mucosal surfaces. oxidized by H(2)O(2), a reaction catalyzed lactoperoxidase, to produce OSCN(-) (hypothiocyanite), molecule with antimicrobial activity. Given the importance availability airway surface fluid, we studied transepithelial transport human bronchial epithelium. We found evidence for at least three mechanisms basolateral apical flux. cAMP and Ca(2+) regulatory pathways controlled through cystic...
TMEM16F is a membrane protein with possible dual function as an ion channel and phospholipid scramblase. The properties of channels associated the link between scramblase activity are matter debate. We studied four isoforms generated by alternative splicing. Upregulation three or silencing endogenous increased decreased, respectively, both activities. Introduction activating mutation in sequence caused marked increase phosphatidylserine scrambling transport indicating direct involvement...
The F508del mutation, the most frequent in cystic fibrosis (CF), impairs maturation of CFTR chloride channel. defect can be partially overcome at low temperature (27°C) or with pharmacological correctors. However, efficacy correctors on mutant protein appears to dependent cell expression system. We have used a bronchial epithelial line, CFBE41o−, determine various known treatments and discover new Compared other types, CFBE41o− shows largest response lowest one such as corr-4a VRT-325. A...
Gene therapy has always been a promising therapeutic approach for Cystic Fibrosis (CF). However, numerous trials using DNA or viral vectors encoding the correct protein resulted in general low efficacy. In last years, chemically modified messenger RNA (cmRNA) proven to be highly potent, pulmonary drug. Consequently, we first explored expression, function and immunogenicity of human (h)CFTR encoded by cmRNAhCFTR vitro ex vivo, quantified expression flow cytometry, determined its YFP based...
Loss-of-function mutations of the CFTR gene cause cystic fibrosis (CF) through a variety molecular mechanisms involving altered expression, trafficking, and/or activity chloride channel. The most frequent mutation among CF patients, F508del, causes multiple defects that can be, however, overcome by combination three pharmacological agents improve channel trafficking and gating, namely, elexacaftor, tezacaftor, ivacaftor. This study was prompted evidence two compound heterozygous for F508del...
We identified a small molecule that, at picomolar concentrations, rescues mutant CFTR chloride channel from protein degradation.
A large fraction of mutations causing cystic fibrosis impair the function transmembrane conductance regulator (CFTR) chloride channel by reduced activity (gating defect) and/or impaired exit from endoplasmic reticulum (trafficking defect). Such defects need to be treated with separate pharmacological compounds termed potentiators and correctors, respectively. Here, we report characterization aminoarylthiazoles (AATs) as having dual activity. Cells expressing mutant CFTR were studied...
The Ca(2+)-activated Cl(-) channels (CaCCs) are involved in a variety of physiological functions, such as transepithelial anion transport, smooth muscle contraction and olfaction. Recently, the question molecular identity CaCCs has apparently been resolved with identification TMEM16A protein (also known anoctamin-1). Expression is associated appearance Ca(2+)- voltage-dependent currents properties similar to those native CaCCs. putative structure consists eight transmembrane domains, both...
Approximately 30% of alleles causing genetic disorders generate premature termination codons (PTCs), which are usually associated with severe phenotypes. However, bypassing the deleterious stop codon can lead to a mild disease outcome. Splicing at NAGNAG tandem splice sites has been reported result in insertion or deletion (indel) three nucleotides. We identified such mechanism as origin asymptomatic phenotype observed cystic fibrosis patients homozygous for E831X mutation (2623G>T) CFTR...
Abstract Cystic fibrosis (CF) is caused by mutations in the CFTR chloride channel. Deletion of phenylalanine 508 (F508del), most frequent CF mutation, impairs trafficking and gating. F508del-CFTR mistrafficking may be corrected acting directly on mutant itself or modulating expression/activity CFTR-interacting proteins, that thus represent potential drug targets. To evaluate possible candidates for rescue, we screened a siRNA library targeting known interactors. Our analysis identified RNF5...
TMEM16A and TMEM16B proteins are CaCCs (Ca2+-activated Cl- channels) with eight putative transmembrane segments. As shown previously, expression of generates characterized by a 10-fold lower Ca2+ affinity faster activation deactivation kinetics respect to TMEM16A. To investigate the basis different properties, we generated chimaeric in which domains protein were replaced equivalent TMEM16B. Replacement N-terminus, TMD (transmembrane domain) 1-2, first intracellular loop TMD3-4 did not change...
Induction of mucus hypersecretion in the airway epithelium by Th2 cytokines is associated with expression TMEM16A, a Ca2+-activated Cl- channel. We asked whether exposure epithelial cells to bacterial components, condition that mimics highly infected environment occurring cystic fibrosis (CF), also results similar response. In cultured human bronchial cells, treatment pyocyanin or P. aeruginosa culture supernatant caused significant increase TMEM16A function. The Ca2+-dependent secretion,...
In cystic fibrosis, deletion of phenylalanine 508 (F508del) in the fibrosis transmembrane conductance regulator (CFTR) anion channel causes misfolding and premature degradation. One possible approach to reducing detrimental health effects could be identification proteins whose suppression rescues F508del-CFTR function bronchial epithelial cells. However, searches for these potential targets have not yet been conducted, particularly a relevant airway background using functional readout. To...
Abstract The advent of Trikafta (Kaftrio in Europe) (a triple-combination therapy based on two correctors—elexacaftor/tezacaftor—and the potentiator ivacaftor) has represented a revolution for treatment patients with cystic fibrosis (CF) carrying most common misfolding mutation, F508del-CFTR. This proved to be great efficacy people homozygous F508del-CFTR and is also useful individuals single F508del allele. Nevertheless, this needs improved, especially light extent its use rare class II...
In cystic fibrosis (CF), deletion of phenylalanine 508 (F508del) in the CF transmembrane conductance regulator (CFTR) is associated to misfolding and defective gating mutant channel. One most promising drug targets ubiquitin ligase RNF5, which promotes F508del-CFTR degradation. Recently, first ever reported inhibitor RNF5 was discovered, i.e., 1,2,4-thiadiazol-5-ylidene inh-2. Here, we designed synthesized a series new analogues explore structure–activity relationships (SAR) this class...
TMEM16A protein, also known as anoctamin-1, has been recently identified an essential component of Ca2+-activated Cl− channels. We previously reported the existence different isoforms generated by alternative splicing. In present study, we have determined functional properties a minimal protein. This isoform, called TMEM16A(0), significantly shortened amino-terminus and lacks three segments localized in intracellular regions protein (total length: 840 amino acids). TMEM16A(0) expression is...
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