A5057718388- Cystic Fibrosis Research Advances
- Neonatal Respiratory Health Research
- Cardiovascular Effects of Exercise
- Cardiac electrophysiology and arrhythmias
- Tracheal and airway disorders
- Cardiomyopathy and Myosin Studies
- Ion channel regulation and function
- Amino Acid Enzymes and Metabolism
- Tissue Engineering and Regenerative Medicine
- Cardiac Arrhythmias and Treatments
- RNA and protein synthesis mechanisms
- Gut microbiota and health
- Renal and related cancers
- Cardiac Valve Diseases and Treatments
- Bone fractures and treatments
- Autism Spectrum Disorder Research
- Asthma and respiratory diseases
- Energy Harvesting in Wireless Networks
- Protein Structure and Dynamics
- Mitochondrial Function and Pathology
- Cardiac Fibrosis and Remodeling
- TGF-β signaling in diseases
- Pluripotent Stem Cells Research
- Microfluidic and Bio-sensing Technologies
- Orthopaedic implants and arthroplasty
University of Toronto
2015-2023
Ted Rogers Centre for Heart Research
2020-2023
Hospital for Sick Children
2015-2021
SickKids Foundation
2018-2019
Molecular Medicine Research Institute
2018
University College London
2018
Great Ormond Street Hospital
2018
Hospital for Special Surgery
2003
Abstract The combination therapy of lumacaftor and ivacaftor (Orkambi ® ) is approved for patients bearing the major cystic fibrosis ( CF mutation: ΔF508 . It has been predicted that Orkambi could treat with rarer mutations similar “theratype”; however, a standardized approach confirming efficacy in these cohorts not reported. Here, we demonstrate rare c.3700 A>G, causing protein misprocessing altered channel function—similar to ΔF508‐ CFTR , are unlikely yield robust response. While...
Pulmonary disease is the major cause of morbidity and mortality in patients with cystic fibrosis, a caused by mutations Cystic Fibrosis Transmembrane conductance Regulator (CFTR) gene. Heterogeneity CFTR genotype-phenotype relationships affected individuals plus escalation drug discovery targeting specific highlights need to develop robust vitro platforms which stratify therapeutic options using relevant tissue. Toward this goal, we adapted fluorescence plate reader assay apical...
Significance Cystic fibrosis is caused by mutations in the cystic transmembrane conductance regulator (CFTR) gene that encodes a chloride channel located apical membrane of epithelia cells. The cAMP signaling pathway and protein phosphorylation are known to be primary controlling mechanisms for function. In this study, we present an alternative activation involves calcium-activated calmodulin binding intrinsically disordered regulatory (R) region CFTR. Beyond their potential therapeutic...
ABSTRACT Cystic fibrosis (CF) is caused by mutations in the CFTR gene and associated with progressive ultimately fatal infectious lung disease. There can be considerable variability disease severity among individuals same mutations, recent genome-wide association studies have identified secondary genetic factors that contribute to this. One of these modifier genes SLC6A14 , which encodes an amino acid transporter. Importantly, variants this been age at first acquisition Pseudomonas...
For those people with cystic fibrosis carrying rare CFTR mutations not responding to currently available therapies, there is an unmet need for relevant tissue models therapy development. Here, we describe a new testing platform that employs patient-specific induced pluripotent stem cells (iPSCs) differentiated lung progenitor can be studied using dynamic, high-throughput fluorescence-based assay of channel activity. Our proof-of-concept studies support the potential use this platform,...
The severity of intestinal disease associated with Cystic Fibrosis (CF) is variable in the patient population and this variability partially conferred by influence modifier genes. Genome-wide association studies have identified SLC6A14, an electrogenic amino acid transporter, as a genetic CF-associated meconium ileus. purpose current work was to determine biological role Slc6a14, disrupting its expression CF mice bearing major mutation, F508del. We found that disruption Slc6a14 worsened...
Abstract The intercalated disc (ICD) is a unique membrane structure that indispensable to normal heart function, yet its structural organization not completely understood. Previously, we showed the ICD-bound transmembrane protein 65 (Tmem65) was required for connexin43 (Cx43) localization and function in cultured mouse neonatal cardiomyocytes. Here, investigate functional cellular effects of Tmem65 reductions on myocardium model by injecting CD1 pups (3–7 days after birth) with recombinant...
SLC6A14-mediated l-arginine transport has been shown to augment the residual anion channel activity of major mutant, F508del-CFTR, in murine gastrointestinal tract. It is not yet known if this transporter augments and pharmacological corrected F508del-CFTR primary airway epithelia. We sought determine role uptake via SLC6A14 modifying cells from patients with cystic fibrosis (CF). Human bronchial epithelial (HBE) lung explants without CF those (CF-HBE) were used for H3-flux, surface liquid,...
In the current study we examined several proteomic- and RNA-Seq-based datasets of cardiac-enriched, cell-surface membrane-associated proteins in human fetal mouse neonatal ventricular cardiomyocytes. By integrating available microarray tissue expression profiles with MGI phenotypic analysis, identified 173 that are conserved amongst eukaryotic species, have not yet been linked to a 'cardiac' Phenotype-Ontology. To highlight utility this dataset, selected investigate more carefully, including...
Abstract Cell lineage conversion of fibroblasts to specialized cell types through transdifferentiation may provide a fast and alternative source for regenerative medicine. Here we show that transient transduction with the four reprogramming factors (Oct4, Sox2, Klf4, c-Myc) in addition early lung transcription factor Nkx2-1 (also known as Ttf1), followed by directed differentiation cells, can convert mouse embryonic human adult dermal into induced lung-like epithelial cells (iLEC). These...
Induced Pluripotent Stem Cells (iPSCs) can be differentiated into epithelial organoids that recapitulate the relevant context for CFTR and enable testing of therapies targeting Cystic Fibrosis (CF)-causing mutant proteins. However, to date, CF-iPSC-derived have only been used study pharmacological modulation channel activity not other disease-relevant membrane protein constituents. In current work, we describe a high-throughput, fluorescence-based assay in iPSC-derived intestinal how this...
Cell-based models that mimic in vivo heart physiology are poised to make significant advances cardiac disease modeling and drug discovery. In these systems, cardiomyocyte (CM) contractility is an important functional metric, but current measurement methods inaccurate low-throughput or require complex setups. To address this need, we developed a standalone noninvasive, label-free ultrasound technique operating at 40-200 MHz measure the contractile kinetics of models, ranging from single adult...
The sarco(endo)plasmic reticulum (SR/ER) is an essential regulator of many key cellular processes, especially those that play a role in the development and progression cardiac disease. However, aspects its structural organization remain poorly defined. Receptor Expression Enhancing Protein 5 (REEP5) enriched SR/ER membrane protein, which regulates highly differentiated network responses to stress. In zebrafish models, genetic knock-out reep5 results functional defects reduced heart rate....
Intercalated discs (ICDs) are unique and functionally indispensible to the heart, but its structural organization remains less understood. Previously, we showed that an ICD-bound transmembrane protein 65 (Tmem65) was required for Connexin 43 (Cx43) localization in cultured mouse neonatal cardiomyocytes, reduced Tmem65 associated with a decrease internalization of Cx43, impaired electrical conduction between neighboring cardiomyocytes. Here, investigated role vivo by injecting CD1 mice...
Fluorescence-based studies are suitable for high-throughput plate reader assays of cells in culture. They have been commonly employed drug discovery campaigns targeting recombinant ion channel proteins overexpressed such as HEK-293 cells. However, there is increasing emphasis on the use tissue-relevant cell lines studying effects small molecule interventions. The following protocol describes adaptation a fluorescence-based membrane potential assay study channels endogenously expressed...
The intercalated disc (ICD) is unique membrane structure that indispensable to normal heart function, yet its structural organization not completely understood. Previously, we showed the ICD-bound transmembrane protein 65 (Tmem65) was required for connexin 43 (Cx43) localization and function in cultured mouse neonatal cardiomyocytes. Here, investigated role of Tmem65 ICD vivo . A model established by injecting CD1 pups (3-7 days after birth) with recombinant adeno-associated virus 9 (rAAV9)...
The intercalated disc (ICD) is unique membrane structure that indispensable to normal heart function. However, its structural organization not well understood. Previously, we showed the ICD-bound transmembrane protein 65 (Tmem65) was required for connexin 43 (Cx43) localization in cultured mouse neonatal cardiomyocytes. Here, investigated role of Tmem65 ICD vivo . A model established by injecting CD1 pups (3-7 days after birth) with recombinant adeno-associated virus 9 (rAAV9) harboring (or...