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Biology subjects

Armstrong, P. C.

Publications and source records attributed to Armstrong, P. C..

3 recordsLinked to original sources

Tissue fibroblasts are a critical source of prostacyclin and anti-thrombotic

Prostacyclin is one of the bodies fundamental signalling pathways. It has long been considered principally anti-thrombotic hormone derived from the vascular endothelium. The role of non-vascular sources in prostacyclin synthesis has not been systematically evaluated, however, due to a lack of tools resulting in an underappreciation of its role in other contexts. Here we used cellspecific knockout mice and human tissues to show that lung, and other tissues, are powerful producers of prostacyclin independent of their vascular components. Instead, in mice and humans, lung prostacyclin synthesis is associated with fibroblasts. The fibroblast-derived prostaglandins enter the circulation and provide systemic anti-thrombotic protection. These observations define a new paradigm in prostacyclin biology in which fibroblast/non-vascular-derived prostacyclin works in parallel with endothelium-derived prostaglandins to control cardiovascular health and potentially a broad range of other biology. These results may explain how local diseases of the lung and elsewhere result in cardiovascular risk.

pharmacology and toxicology↗

Temporal in vivo platelet labelling in mice reveals age-dependent receptor expression and conservation of specific mRNAs

The proportion of young platelets, also known as newly formed or reticulated, within the overall platelet population has been clinically correlated with adverse cardiovascular outcomes. Our understanding of this is incomplete, however, because of limitations in the technical approaches available to study platelets of different ages. In this study we have developed and validated an in vivo temporal labelling approach using injectable fluorescent anti-platelet antibodies to sub-divide platelets by age and assess differences in functional and molecular characteristics. With this approach we found that young platelets (<24h old) in comparison to older platelets respond to stimuli with greater calcium flux and degranulation, and contribute more to the formation of thrombi in vitro and in vivo. Sequential sampling confirmed this altered functionality to be independent of platelet size with no size differences or changes relative to the global population seen at any age. The age associated decrease in thrombotic function was accompanied by significant decreases in the surface expression of GPVI and CD31 (PECAM-1) and an increase in CD9. Platelet mRNA content also decreased with age but at different rates for individual mRNAs indicating apparent conservation of those encoding granule proteins. Our pulse-chase type approach to define circulating platelet age has allowed timely re-examination of commonly held beliefs regarding size and reactivity of young platelets whilst providing novel insights into the temporal regulation of receptor and protein expression. Overall, future application of this validated tool will inform on age-based platelet heterogeneity in physiology and disease.

physiology↗

Proteome and functional decline as platelets age in the circulation

Anucleate platelets circulate in the blood of healthy individuals for approximately 7-10 days during which time their protein composition may change. We hypothesized such changes would be linked to altered structure and function. Here, we separated platelets of different ages based on mRNA content and characterised them using proteomics, immunofluorescence and functional assays. Total protein content was 45{+/-}5% (n=4) lower in old platelets compared to young platelets. Predictive proteomic pathway analysis identified associations with 28 biological processes, notably increased haemostasis in young platelets and apoptosis in old platelets. Further studies confirmed platelet ageing was linked to a reduction decrease in cytoskeletal proteins, a reduction in mitochondria number, and lower calcium dynamics and granule secretion. This work delineates physical and functional changes in platelets as they age and serves as a base to examine differences associated with altered mean age of platelet populations in conditions such as immune thrombocytopenia and diabetes.

cell biology↗