Characterizing mouse platelet heterogeneity across diverse disease models using spectral flow cytometry and high-dimensional analysis.

Gautam, D., Clarke, E. M., Zon, R. L., Smith-Oliver, M. R., Kumar, A., Sullivan, M. E., Karagiannis, P., Roweth, H. G. ORCID: https://orcid.org/0000-0002-1100-8409 and Battinelli, E. M. (2026) Characterizing mouse platelet heterogeneity across diverse disease models using spectral flow cytometry and high-dimensional analysis. Research and practice in thrombosis and haemostasis, 10 (2). 103371. ISSN 2475-0379 doi: 10.1016/j.rpth.2026.103371

Abstract/Summary

BACKGROUND Routine platelet assessment based on count and mean platelet volume overlooks heterogeneity of platelet subpopulations that influence disease outcomes. Different platelet subtypes are associated with diverse pathological conditions, highlighting the need to define and characterize them. Human studies face limitations due to interindividual variability and challenges in acquiring matched controls. Moreover, the small and anucleate nature of platelets constrain conventional single-cell analysis approaches. These gaps highlight the need for a mouse-specific flow cytometry panel to enable detailed investigation of platelet heterogeneity in preclinical models. OBJECTIVES To develop and validate a mouse-specific spectral flow cytometry panel integrated with a high-dimensional analysis pipeline for comprehensive characterization of platelet subpopulations and activation states under physiological and pathological conditions. METHODS A 12-marker spectral panel was optimized and integrated with the PlateletProfiler pipeline for multidimensional clustering and receptor expression profiling. The workflow was applied to conditions known to alter platelet dynamics, including agonist-induced activation and three mouse models of disease: lipopolysaccharide-induced inflammation, Jak2V617F driven myeloproliferative neoplasms, and breast cancer. RESULTS Four major platelet subpopulations-resting, primed, aggregatory, and procoagulant-were identified, representing a continuum of activation. Lipopolysaccharide exposure increased primed and aggregatory subsets, Jak2V617F mice showed aggregatory and procoagulant fractions, and tumor-bearing mice exhibited increased procoagulant platelets. Across models, platelets displayed upregulation of activation and procoagulant markers. All disease models displayed elevated thiazole orange-positive reticulated platelets. CONCLUSIONS This integrated and scalable workflow provides a robust platform for investigating disease-associated changes in platelet heterogeneity. The PlateletProfiler pipeline is compatible with both mouse and human datasets, supporting broad experimental and translational applications.

Item Type	Article
URI	https://centaur.reading.ac.uk/id/eprint/129711
Identification Number/DOI	10.1016/j.rpth.2026.103371
Refereed	Yes
Divisions	Life Sciences > School of Biological Sciences > Biomedical Sciences
Publisher	Elsevier
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