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Howell, O.

Publications and source records attributed to Howell, O..

2 recordsLinked to original sources

Spatial transcriptomics of compartmentalised inflammation in a natural disease multiple sclerosis cohort.

Compartmentalised inflammation is a poorly understood aspect of multiple sclerosis (MS) that is associated with worse outcomes and represents an important therapeutic target. To gain deeper insight into compartmentalised inflammation, we have taken the approach of digital spatial profiling of the whole human transcriptome in areas of perivascular and meningeal inflammation and tertiary lymphoid-like structures (TLS) in MS central nervous system tissue. Critically, we had access to rare archival tissue obtained before the era of disease-modifying therapies, representing a natural history of disease. This analysis has identified differentially expressed genes in TLS compared to meningeal or perivascular inflammation. Pathway analysis highlighted that TLS signalling is dominated by B cell activity including active antibody secretion. Our data demonstrated the diversity of immunoglobulins and the prominence of IgG3- and IgG4-secreting cells in TLS. Intriguingly, pathway analysis suggests TLS may be hubs for viral (re)activation which warrants further investigation. These findings provide insight into the function of TLS in MS disease pathogenesis and reveal unique immune signatures that may support biomarker development to predict which patients harbour TLS in life.

neuroscience↗

Imaging Oxysterols in Mouse Brain by On-Tissue Derivatisation-Robotic Liquid Micro-Extraction Surface Analysis-Liquid Chromatography Mass Spectrometry

Dysregulated cholesterol metabolism is implicated in a number of neurological disorders. Many sterols, including cholesterol and its precursors and metabolites, are biologically active and important for proper brain function. However, spatial cholesterol metabolism in brain and the resulting sterol distributions are poorly defined. To better understand cholesterol metabolism in situ across the complex functional regions of brain, we have developed on-tissue enzyme-assisted derivatisation in combination with micro-liquid-extraction for surface analysis and liquid chromatography - mass spectrometry to image sterols in tissue slices (10 {micro}m) of mouse brain. The method provides sterolomic analysis at 400 {micro}m spot diameter with a limit of quantification of 0.01 ng/mm2. It overcomes the limitations of previous mass spectrometry imaging techniques in analysis of low abundance and difficult to ionise sterol molecules, allowing isomer differentiation and structure identification. Here we demonstrate the spatial distribution and quantification of multiple sterols involved in cholesterol metabolic pathways in wild type and cholesterol 24S-hydroxylase knock-out mouse brain. The technology described provides a powerful tool for future studies of spatial cholesterol metabolism in healthy and diseased tissues.\n\nSignificanceThe brain is a remarkably complex organ and cholesterol homeostasis underpins brain function. It is known that cholesterol is not evenly distributed across different brain regions, however, the precise map of cholesterol metabolism in the brain remains unclear. If cholesterol metabolism is to be correlated with brain function it is essential to generate such a map. Here we describe an advanced mass spectrometry imaging platform to reveal spatial cholesterol metabolism in situ at 400 {micro}m resolution on 10 {micro}m tissue slices from mouse brain. We mapped, not only cholesterol, but also other biologically active sterols arising from cholesterol turnover in both wild type and mice lacking cholesterol 24-hydroxylase (Cyp46a1), the major cholesterol metabolising enzyme.

neuroscience↗