Project description:European-American individuals of the GENOA cohort participating in the “Genetics of Microangiopathic Brain Injury” substudy, which investigates the genetic basis of alteration in brain structure detectable by magnetic resonance imaging. This analysis investigated the association of gene expression with age (at the time of cell transformation).
Project description:An H5N1 virus-encoded microRNA directly targets mammalian poly(rC) binding protein 2 and is a major contributor to H5N1-associated ‘cytokine storm’ and mortality.
Project description:Here we introduce STORM-seq, a standardized and generally available approach to profiling total RNA in single cells by miniaturization of the Takara SMART-seq Stranded (SSS) kit. The use of nanoliter-scale reaction volumes allows STORM-seq to efficiently scale to hundreds of high-complexity single cell libraries per run, from dissociated tissues and non-adherent cells, with greater than 99% per cell/library success. The use of ribosomal RNA depletion and random hexamer priming, rather than oligo-dT (poly-A) selection, allows comprehensive, whole-transcriptome analysis in each individual cell. STORM-seq recovers both coding and non-coding transcripts at an equal or greater rate to full volume reactions. Applying the method to cultured K562 cells and primary human fallopian tube epithelial (FTE) cells results in equivalent or better gene detection than manual preparation. Across all transcript biotypes, additional mRNA and lncRNA transcripts are detected beyond SSS or SMART-seq2. STORM-seq libraries from primary human FTE reveal intermediate cell types not observed by other technologies, with clear transitional significance visible upon data integration. STORM-seq provides a scalable, efficient protocol to recover deep whole transcriptomes from individual cells while minimizing costs.
Project description:SARS-CoV2 infection leads to cardiac injury and dysfunction in 20-30% of hospitalized patients and higher rates of mortality in patients with pre-existing cardiovascular disease. Inflammatory factors released as part of the 'cytokine storm' are thought to play a critical role in cardiac dysfunction in severe COVID-19 patients. Here we use human cardiac organoid technology combined with high sensitivity phosphoproteomics and single nuclei RNA sequencing to identify inflammatory targets inducing cardiac dysfunction. This new pipeline allowed rapid progress and identification of putative therapeutics. We identify a novel interferon-gamma driven BRD4 (bromodomain protein 4)-fibrosis/iNOS axis as a key intracellular mediator of inflammation-induced cardiac dysfunction. This axis is therapeutically targetable using BRD4 inhibitors, which promoted full recovery of function in human cardiac organoids and prevented severe inflammation and death in a cytokine-storm mouse model. The BRD inhibitor INCB054329 was the most efficacious, and is a prime candidate for drug repurposing to attenuate cardiac dysfunction and improve COVID-19 mortality in humans.
Project description:This study examines the best way to teach genetics to family medicine residents. First year family medicine residents at the University of Toronto will be taught basic clinical genetics as well as a specific disease in genetics via 3 different educational methods. All participants will undergo an oral examination and written knowledge test 3 months after this education. Results between groups will be compared, and the best way to teach genetics to residents determined.
Project description:European-American individuals of the GENOA cohort participating in the “Genetics of Microangiopathic Brain Injury” substudy, which investigates the genetic basis of alteration in brain structure detectable by magnetic resonance imaging. This analysis investigated the association of gene expression with age (at the time of cell transformation). Participants in this study are drawn from the GENOA study, a population-based study from Rochester, MN
