A5052055623- Cystic Fibrosis Research Advances
- Neonatal Respiratory Health Research
- Asthma and respiratory diseases
- Tracheal and airway disorders
- Respiratory viral infections research
- Virus-based gene therapy research
- Advanced biosensing and bioanalysis techniques
- Ion Transport and Channel Regulation
- Inhalation and Respiratory Drug Delivery
- Child Nutrition and Feeding Issues
- Congenital Ear and Nasal Anomalies
- Viral gastroenteritis research and epidemiology
- Olfactory and Sensory Function Studies
- Endoplasmic Reticulum Stress and Disease
- Immunodeficiency and Autoimmune Disorders
- Respiratory and Cough-Related Research
- Ion channel regulation and function
- RNA Interference and Gene Delivery
- Pediatric health and respiratory diseases
- Infant Nutrition and Health
- Advanced Biosensing Techniques and Applications
- Dysphagia Assessment and Management
- Legume Nitrogen Fixing Symbiosis
- Biochemical Analysis and Sensing Techniques
- Biosensors and Analytical Detection
National Jewish Health
2018-2024
University of Colorado Denver
2003-2024
University of Colorado Anschutz Medical Campus
2020-2022
Denver School of Nursing
2019-2022
The Medical Center of Aurora
2022
University of Virginia
2022
Johns Hopkins University
2008-2018
Johns Hopkins Medicine
2007-2017
University of Baltimore
2014
Kennedy Krieger Institute
2009
Adeno-associated virus (AAV) vectors expressing the normal cystic fibrosis transmembrane conductance regulator (CFTR) cDNA complement (CF) defect in vitro. Unlike other DNA vectors, AAV is a stably integrating virus, which could make possible long-term vivo complementation of CF airway epithelium. We report AAV-CFTR gene transfer and expression after infection primary nasal polyp cells delivery vector to one lobe rabbit lung through fiberoptic bronchoscope. In rabbit, be detected infected up...
Protective mucus coatings typically trap and rapidly remove foreign particles from the eyes, gastrointestinal tract, airways, nasopharynx, female reproductive thereby strongly limiting opportunities for controlled drug delivery at mucosal surfaces. No synthetic system composed of biodegradable polymers has been shown to penetrate highly viscoelastic human mucus, such as non-ovulatory cervicovaginal a significant rate. We prepared nanoparticles diblock copolymer poly(sebacic acid)...
Antisera against two peptides, corresponding to different domains of the cystic fibrosis gene product CFTR, have been raised and extensively characterized. Both antisera recognize CFTR as a 165-kDa polypeptide in Western analysis cells transfected with cDNA well epithelial cell lines. The tissue distribution has studied by immunocytochemistry. is abundant cells, including those lining sweat ducts, small pancreatic intestinal crypts. Unexpectedly, level lung epithelia relatively low, while it...
The most common cystic fibrosis transmembrane conductance regulator mutation, delta F508-CFTR, is a partially functional chloride channel that retained in the endoplasmic reticulum and degraded. We hypothesize known transcriptional regulator, sodium 4-phenylbutyrate (4PBA), will enable greater fraction of F508-CFTR to escape degradation appear at cell surface. Primary cultures nasal polyp epithelia from CF patients (delta F508 homozygous or heterozygous), bronchial epithelial line IB3-1...
Our objectives were to characterise the microbiota in cystic fibrosis (CF) bronchoalveolar lavage fluid (BALF), and determine its relationship inflammation disease status. BALF from paediatric adult CF patients controls undergoing clinically indicated bronchoscopy was analysed for total bacterial load by 16S rDNA sequencing. We examined 191 samples (146 45 controls) 13 centres. In aged <2 years, nontraditional taxa ( e.g . Streptococcus , Prevotella Veillonella ) constituted ∼50% of...
Adeno-associated virus type 2 (AAV) vectors have been used for gene expression in respiratory epithelial cells and may be useful therapy diseases like cystic fibrosis (CF) which affect the airways. The AAV p5 promoter together with inverted terminal repeat (ITR) forms a 263-base pair cassette mediated efficient CF bronchial cell line. We report here that ITR itself can mediate expression. In stable transfection assays, AAV-CF expressing full-length transmembrane conductance regulator (CFTR)...
Sodium 4-phenylbutyrate (Buphenyl, 4PBA) is a new FDA approved drug for management of urea cycle disorders. We have previously presented data suggesting that 4PBA, at clinically achievable concentrations, induces CFTR channel function on the plasma membrane Δ F508-expressing cystic fibrosis (CF) airway epithelial cells in vitro (Rubenstein, R. C., and P. L. Zeitlin, 1997. J. Clin. Invest. 100:2457– 2463). hypothesized 4PBA would induce vivo individuals homozygous F508-CFTR. A randomized,...
We conducted a double-blind, placebo-controlled, multicenter, randomized trial to test the hypothesis that 300 mg of tobramycin solution for inhalation administered twice daily 28 days would be safe and result in profound decrease Pseudomonas aeruginosa (Pa) density from lower airway young children with cystic fibrosis. Ninety-eight subjects were randomized; however, was stopped early because evidence significant microbiological treatment effect. Twenty-one under age 6 years (8 active; 13...
An immortalized cell line was created from a primary culture of bronchial epithelia isolated patient with cystic fibrosis. The transformed hybrid virus, adeno-12-SV40, which has been used successfully on number different human epithelial tissues. cells have the following characteristics. (1) Cyclic adenosine monophosphate (cAMP) is stimulated by beta-adrenergic agonists. (2) Outwardly rectifying Cl- channels are present apical membrane. These can be activated depolarizing voltages but not...
Secretory chloride channels can be activated by adenosine 3′,5′-monophosphate (cAMP)-dependent protein kinase in normal airway epithelial cells but not from individuals with cystic fibrosis (CF). In excised, inside-out patches of apical membrane human and three patients CF, the exhibited a characteristic outwardly rectifying current-voltage relation depolarization-induced activation. Channels tissues were both cAMP-dependent C. However, CF could either kinase. Thus, gating is regulated C,...
The most common mutation of the cystic fibrosis transmembrane conductance regulator (CFTR), ΔF508, is a trafficking mutant that has prolonged associations with molecular chaperones and rapidly degraded, at least in part by ubiquitin-proteasome system. Sodium 4-phenylbutyrate (4PBA) improves ΔF508-CFTR function vitro epithelial cells vivo. To further understand mechanism action 4PBA, we tested hypothesis 4PBA modulates targeting for ubiquitination degradation reducing expression Hsc70 cells....
Previous studies have demonstrated that delivery of a recombinant adeno-associated virus (AAV) vector encoding the complete human cystic fibrosis transmembrane regulator (CFTR) cDNA (tgAAVCF) to nose, sinus, and lungs subjects with (CF) was safe well tolerated. In small randomized, double-blind study three doses aerosolized tgAAVCF or placebo at 30-day intervals, encouraging but non-significant trends in pulmonary function induced sputum interleukin 8 (IL-8) levels were seen early time...
Recombinant adeno-associated serotype 2-based vectors (rAAV2) possess a number of theoretical advantages for cystic fibrosis (CF) gene therapy because they elicit little or no inflammatory response and generally result in stable expression. rAAV2 expressing the transmembrane conductance regulator (CFTR) have previously been shown to mediate correction CF defect bronchial epithelial cells expression CFTR rabbit nonhuman primate models. Here we report results first trial initiated with rAAV...
Adeno-associated virus type 2 (AAV2)-based vectors are capable of stable expression in the airway epithelium and may be useful for gene therapy human diseases, such as cystic fibrosis. Certain vectors, retroviruses, require active cell division integration expression, but this has not been formally evaluated case AAV2. The fibrosis bronchial epithelial line, IB3-1, which can transduced by AAV2 was shown to undergo a decrease DNA synthesis undetectable levels when grown confluence. Cultures <...
Most individuals with cystic fibrosis (CF) carry one or two mutations that result in a maturation defect of the full-length protein. One such mutation, ΔF508, results mutant membrane glycoprotein fails to progress apical membrane, where wild-type protein normally functions as cyclic AMP-regulated chloride channel. 4-Phenylbutyrate (Buphenyl), an orally bioavailable short chain fatty acid, modulates heat shock expression and restores ΔF508 vitro vivo. We performed randomized, double-blind,...
Endoplasmic reticulum (ER)-associated degradation (ERAD) is the major quality control pathway of cell. The most common disease-causing protein folding mutation, DeltaF508-cystic fibrosis transmembrane regulator (CFTR), destroyed by ERAD to cause cystic (CF). p97/valosin-containing (VCP) physically interacts with gp78/autocrine motility factor receptor couple ubiquitination, retrotranslocation, and proteasome misfolded proteins. We show here that p97/VCP gp78 form complexes CFTR during...
Background Dysfunctional CFTR in the airways is associated with elevated levels of NFκB mediated IL-8 signaling leading to neutrophil chemotaxis and chronic lung inflammation cystic fibrosis. The mechanism(s) by which mediates inflammatory under debate. Methodology/Principal Findings We tested hypothesis that wt-CFTR down-regulates secretion. transiently co-expressed or promoters driving luciferase expression HEK293 cells. Wt-CFTR cells suppresses both basal IL1β induced IL-8, promoter...
Abstract Cystic fibrosis (CF) is caused by mutations in the cystic transmembrane conductance regulator (CFTR), which lead to early death due progressive lung disease. The development of small-molecule modulators that directly interact with CFTR aid protein folding (“correctors”) and/or increase channel function (“potentiators”) have proven be highly effective therapeutic treatment CF. Notably, incorporation next-generation corrector, elexacaftor, into a triple combination (marketed as...
Cystic fibrosis is a life-limiting autosomal recessive disorder with highly variable clinical presentation. The classic form involves characteristic findings in the respiratory tract, gastrointestinal male reproductive and sweat glands caused by loss-of-function mutations cystic transmembrane conductance regulator (CFTR ) gene. Nonclassic forms of have been associated that reduce but do not eliminate function CFTR protein. We assessed whether alteration responsible for entire spectrum...
Induced sputum has been used to study airway inflammation. We sought determine whether markers of infection and inflammation in induced were a useful safe outcome measure cystic fibrosis. hypothesized that bacterial density inflammatory content would decrease after antibiotic therapy. was assayed for density, cell count, differential before treatment with intravenous antibiotics. Fifty-five the 72 subjects enrolled (mean age ± SD 18.2 7.9 years) completed study. FEV1 increased by an average...