Project description:The diversity of cell types and regulatory states in the brain, and how these change during ageing, remains largely unknown. We present a single-cell transcriptome atlas of the entire adult Drosophila melanogaster brain sampled across its lifespan. Cell clustering identified 87 initial cell clusters that are further subclustered and validated by targeted cell-sorting. Our data shows high granularity and identifies a wide range of cell types. Gene network analyses using SCENIC revealed regulatory heterogeneity linked to energy consumption. During ageing, RNA content declines exponentially without affecting neuronal identity in old brains. This single-cell brain atlas covers nearly all cells in the normal brain and provides the tools to study cellular diversity alongside other Drosophila and mammalian single-cell datasets in our unique single-cell analysis platform. These results allow comprehensive exploration of all transcriptional states of an entire ageing brain.
Project description:We aim to improve anti-ageing drug discovery, currently achieved through laborious and lengthy longevity analysis. Recent studies demonstrated that the most accurate molecular method to measure human age is based on CpG methylation profiles, as exemplified by several epigenetics clocks that can accurately predict an individual’s age. Here, we developed CellAge, a new epigenetic clock that measures subtle ageing changes in primary human cells in vitro. As such it provides a unique tool to measure effects of relatively short pharmacological treatments on ageing. We validated our CellAge clock against known longevity drugs such as rapamycin and trametinib. Moreover, we uncovered novel anti-ageing drugs, torin2 and Dactolisib (BEZ-235), demonstrating the value of our approach as a screening and discovery platform for anti-ageing strategies. CellAge outperforms other epigenetic clocks in measuring subtle ageing changes in primary human cells in culture. The tested drug treatments reduced senescence and other ageing markers, further consolidating our approach as a screening platform. Finally, we showed that the novel anti-ageing drugs we uncovered in vitro, indeed increased longevity in vivo. Our method expands the scope of CpG methylation profiling from measuring human chronological and biological age from human samples in years, to accurately and rapidly detecting anti-ageing potential of drugs using human cells in vitro, providing a novel accelerated discovery platform to test sought after geroprotectors.
Project description:Heart failure is one of the leading causes of death in an ageing population. Hallmarks are cardiac hypertrophy, fibrosis and inflammation. The molecular mechanisms, however, are poorly understood. Glycosylation is one of the most common posttranslational modifications of proteins, which can have important consequences for protein folding, function and turnover. We hypothesized that changes in glycoprotein abundance and glycosylation patterns may contribute to cardiac aging. Western Blot analysis suggests increased protein mannosylation in the aging heart. Glycoprotein pull-downs from heart lysates of young (3 months) and old (2 years) mice in combination with quantitative mass spectrometry support widespread alterations of the glycoproteome in aged hearts.
Project description:Epigenetic modifications including DNA methylation regulate gene expression and contribute to specialization of cells. Currently, there is limited knowledge of the epigenetic patterns of human heart. We have used healthy cardiac tissues from different anatomical regions of the human heart to study unique methylation signatures specific to human heart.