A5027433420- Mitochondrial Function and Pathology
- RNA modifications and cancer
- Cancer, Hypoxia, and Metabolism
- Ubiquitin and proteasome pathways
- Advanced Proteomics Techniques and Applications
- Glycosylation and Glycoproteins Research
- Monoclonal and Polyclonal Antibodies Research
- Ferroptosis and cancer prognosis
- Glioma Diagnosis and Treatment
- Protein Degradation and Inhibitors
- Epigenetics and DNA Methylation
- Cancer-related molecular mechanisms research
- Mass Spectrometry Techniques and Applications
- Cancer Genomics and Diagnostics
- Multiple Myeloma Research and Treatments
- Cancer, Lipids, and Metabolism
- Medical Imaging Techniques and Applications
- HER2/EGFR in Cancer Research
- Cardiac electrophysiology and arrhythmias
- Ion channel regulation and function
- Metabolomics and Mass Spectrometry Studies
- Medicinal Plant Pharmacodynamics Research
- Protein Kinase Regulation and GTPase Signaling
- Radiomics and Machine Learning in Medical Imaging
- Metabolism and Genetic Disorders
Hospital for Sick Children
2013-2023
SickKids Foundation
2008-2022
Great Ormond Street Hospital
2022
University College London
2022
University of Toronto
2008-2020
Princess Margaret Cancer Centre
2008-2014
Institute of Clinical Research
2012
Jiangsu University
2012
Thermo Fisher Scientific (Canada)
2010
McLaughlin Research Institute
2009
The ubiquitin system regulates virtually all aspects of cellular function. We report a method to target the myriad enzymes that govern ubiquitination protein substrates. used massively diverse combinatorial libraries variants develop inhibitors four deubiquitinases (DUBs) and analyzed DUB-inhibitor complexes with crystallography. extended selection strategy conjugating (E2) ligase (E3) found can also enhance enzyme activity. Last, we showed bind selectively ubiquitin-binding domains....
Malignant hyperthermia (MH) and central core disease (CCD) are autosomal dominant disorders of skeletal muscle in which a potentially fatal hypermetabolic crisis can be triggered by commonly used anesthetic agents. To date, 17 mutations the humanRYR1 gene encoding Ca2+ release channel sarcoplasmic reticulum (the ryanodine receptor) have been associated with MH and/or CCD. Although many these linked to CCD, high lod (log odds favoring linkage versus nonlinkage) scores, others found single,...
Central core disease is a rare, nonprogressive myopathy that characterized by hypotonia and proximal muscle weakness. In large Mexican kindred with an unusually severe highly penetrant form of the disorder, DNA sequencing identified I4898T mutation in C-terminal transmembrane/luminal region RyR1 protein constitutes skeletal ryanodine receptor. All previously reported RYR1 mutations are located either cytoplasmic N terminus or central 5,038-aa protein. The was introduced into rabbit cDNA...
Malignant hyperthermia (MH) and central core disease (CCD) mutations were introduced into full-length rabbit Ca2+ release channel (RYR1) cDNA, which was then expressed transiently in HEK-293 cells. Resting concentrations higher cells expressing homotetrameric CCD mutant RyR1 than MH RyR1. Cells or exhibited lower maximal peak amplitudes of caffeine-induced wild type RyR1, suggesting that mutants might be "leaky." In the amplitude 10 mm correlated significantly with carbachol-...
Aberrant expression, activation, and stabilization of epidermal growth factor receptor (EGFR) are causally associated with several human cancers. Post-translational modifications protein-protein interactions directly modulate the signaling trafficking EGFR. Activated EGFR is internalized by endocytosis then either recycled back to cell surface or degraded in lysosome. internalization recycling also occur response stresses that activate p38 MAP kinase. Mass spectrometry was applied...
Abstract Non-small cell lung cancer (NSCLC) is the leading cause of deaths worldwide. Only a fraction NSCLC harbor actionable driver mutations and there an urgent need for patient-derived model systems that will enable development new targeted therapies. other cancers display profound proteome remodeling compared to normal tissue not predicted by DNA or RNA analyses. Here, we generate 137 xenografts (PDXs) recapitulate histology molecular features primary NSCLC. Proteome analysis PDX models...
Ligand binding to the receptor tyrosine kinase fibroblast growth factor (FGF) 1 (FGFR1) causes dimerization and activation by transphosphorylation of residues in domain. FGFR1 is ubiquitylated E3 ligase NEDD4 (also known as NEDD4-1), which promotes internalization degradation. Although phosphorylation required for NEDD4-dependent endocytosis, directly binds a nonphosphorylated region FGFR1. We found that led c-Src kinase-dependent NEDD4, enhancing ubiquitin activity NEDD4. Using mass...
Availability of lung cancer models that closely mimic human tumors remains a significant gap in research, as tumor cell lines and mouse may not recapitulate the spectrum heterogeneity seen patients. We aimed to establish patient-derived xenograft (PDX) resource from surgically resected non-small (NSCLC). Fresh tissue surgical resection was implanted grown subcutaneous pocket non-obese severe combined immune deficient (NOD SCID) gamma mice. Subsequent passages were NOD SCID A subset matched...
In most solid tumors, the Hippo pathway is inactivated through poorly understood mechanisms that result in activation of transcriptional regulators, YAP and TAZ. Here, we identify NUAK2 as a YAP/TAZ activator directly inhibits LATS-mediated phosphorylation show induction by AP-1 required for robust signaling. Pharmacological inhibition or loss reduces growth cultured cancer cells mammary tumors mice. Moreover, human patient samples, expression elevated aggressive, high-grade bladder strongly...
Abstract The deubiquitinase USP5 stabilizes c-Maf, a key transcription factor in multiple myeloma (MM), but the mechanisms and significance are unclear. In present study, was found to interact with c-Maf prevented it from degradation by decreasing its polyubiquitination level. Specifically, 308th 347th lysine residues were critical for USP5-mediated deubiquitination stability. There five domains protein subsequent studies revealed that cryptic ZnF domain C-box interacted UBA1/UBA2 partly...
The balance between comprehensively analyzing the proteome and using valuable mass spectrometry time is a genuine challenge in field of proteomics. Multidimensional fractionation strategies have significantly increased coverage, but often at cost analysis time, despite advances spectrometer acquisition rates. Recently, Evosep One liquid chromatography system was shown to analyze peptide samples high-throughput manner without sacrificing in-depth proteomics coverage. We demonstrate...
Cellular identity in metazoan organisms is frequently established through lineage-specifying transcription factors, which control their own expression transcriptional positive feedback, while antagonizing the developmental networks of competing lineages. Here, we have uncovered a distinct feedback loop that arises from reciprocal stabilization tyrosine kinase ABL and coactivator TAZ. Moreover, determined this required for osteoblast differentiation embryonic skeletal formation. potentiated...
Aberrant expression, activation, and down-regulation of the epidermal growth factor receptor (EGFR) have causal roles in many human cancers, post-translational modifications including phosphorylation ubiquitination protein-protein interactions directly modulate EGFR function. Quantitative mass spectrometric analyses selected reaction monitoring (also known as multiple monitoring) were applied to associated proteins. In response (EGF) stimulation cells, phosphorylations at Ser(991) Tyr(998)...
The capacity to coordinate environmental sensing with initiation of cellular responses underpins microbial survival and is crucial for virulence stress in pathogens. Here we define circuitry that enables the fungal pathogen Candida albicans couple cell cycle dynamics wall induced by echinocandins, a front-line class antifungal drugs. We discover C. transcription factor Cas5 proper which inhibit β-1,3-glucan synthesis. has distinct transcriptional targets under basal conditions, activated...
Glioblastoma is the most common primary brain tumor in adults. While introduction of temozolomide chemotherapy has increased long-term survivorship, treatment failure and rapid recurrence remains universal. The transcriptional regulatory protein, inhibitor DNA-binding-1 (ID1), a key regulator cell phenotype cancer. We show that CRISPR-mediated knockout ID1 glioblastoma cells, breast adenocarcinoma melanoma cells dramatically reduced progression all three cancer systems through downregulation...
The stoichiometry of protein phosphorylation at specific amino acid sites may be used to infer on the significance modification, and its biological function in cell. However, detection quantification tissue remain a significant challenge. Here we describe strategy for highly sensitive, label-free stoichiometry. Method development included analysis synthetic peptides order determine constants relate mass spectrometry signals cognate peptide/phosphopeptide pairs, by using high resolution...
Coiled-coil-helix-coiled-coil-helix domain-containing 2, a mitochondrial protein, encoded by CHCHD2 is located at chromosome 7p11.2 and proximal to the EGFR gene. Here, bioinformatic analyses revealed that consistently coamplified with in non-small cell lung carcinoma (NSCLC). In addition, protein expression levels were positively correlated upregulated relative normal NSCLC tumor-derived xenografts. Knockdown of cells attenuated proliferation, migration, respiration. protein-protein...
UBE2O is proposed as a ubiquitin-conjugating enzyme, but its function was largely unknown. Mass spectrometry applied to identify c-Maf ubiquitination-associated proteins. Immunoprecipitation for and interaction. Immunoblotting used Maf protein stability. Luciferase assay transcriptional activity. Lentiviral infections were in multiple myeloma (MM) cells. Flow cytometry nude mice xenografts MM cell apoptosis tumor growth assay, respectively. found interact with c-Maf, critical transcription...