Real-time monitoring techniques and simplistic platelet assays to reduce and refine animal use in cardiovascular and respiratory biomedical research

Platelets are critical participants in haemostasis, thrombosis and inflammation. We have developed technology for assessing radiolabelled platelet aggregation responses in real-time in anaesthetised mice. Our model is a refinement of thromboembolic mortality models that operates at a lower severity level and uses considerably fewer mice. Mortality models continue to be used by a number of research groups and by initiating dialogue with users we have identified a need for more simplistic alternatives and proof of principle using current test reagents to facilitate uptake. We shall therefore evaluate systemic platelet counting as an alternative to thromboembolic mortality models using conventional antithrombotics and reagents from groups currently using mortality techniques. Our approach will refine procedures currently being used to reduce pain and suffering and will also reduce animal use by participating groups by 50%.

We shall also develop techniques for assessing platelet recruitment to the lung during allergic inflammation in real-time in anaesthetised mice. This will provide an alternative to killing of mice at numerous time points for histological determination of cell infiltration. We anticipate a reduction in mouse use of 70-80%. Antithrombotic drugs will be used to distinguish the inflammatory platelet response from thrombotic platelet activation. The model will ultimately be established in a laboratory that is active in the field of inflammatory lung disease to facilitate immediate application and subsequent dissemination to other research groups.

We therefore propose to develop low cost and easily applied models for assessing platelet activation during cardiovascular and respiratory diseases that refine and reduce procedures involving experimental mice.

Cleary SJ et al. (2019).  Radiolabelling and immunohistochemistry reveal platelet recruitment into lungs and platelet migration into airspaces following LPS inhalation in mice. Journal of Pharmacological and Toxicological Methods 106660. doi: 10.1016/j.vascn.2019.106660

Taylor KA et al. (2019). Pharmacological impact of antiretroviral therapy on platelet function to investigate HIV-associated cardiovascular risk. British Journal of Pharmacology 176(7):879-889. doi: 10.1111/bph.14589

Gillespie S et al. (2018). The isothiocyanate sulforaphane modulates platelet function and protects against cerebral thrombotic dysfunction. British Journal of Pharmacology 175(16):3333-3346. doi: 10.1111/bph.14368

Taylor KA and Emerson M (2018). Refinement of a mouse cardiovascular model: Development, application and dissemination. F1000Research 7:593 doi: 10.12688/f1000research.14456.1

Rauzi F, Smyth E, Emerson M (2017). Refinement of Mouse Protocols for the Study of Platelet Thromboembolic Responses In VivoThrombosis and Haemostasis 117(12): 2283-2290. doi: 10.1160/TH17-04-0250


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Imperial College London

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Award date

Nov 2014 - Mar 2017

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