Journal article
Pyruvate kinase M2 promotes venous thrombosis by enhancing SNAP23-mediated platelet exocytosis and consequent NETosis
Blood advances, Vol.10(1), pp.132-142
01/13/2026
DOI: 10.1182/bloodadvances.2025017414
PMCID: PMC12804119
PMID: 41118613
Abstract
Little is known about the role of metabolic regulatory mechanisms in the pathobiology of deep vein thrombosis (DVT). Recent studies have demonstrated the involvement of the metabolic enzyme pyruvate kinase M2 (PKM2) in platelet function; however, whether platelet PKM2 contributes to DVT has not been investigated yet. Using platelet-specific PKM2-/- (PKM2Plt-KO) or wild-type (WT) mice orally administered ML265 (a small molecule that limits PKM2 dimers by stabilizing PKM2 tetramers), we found reduced thrombus burden at 48 h post-surgery in the inferior vena cava (IVC) stenosis model compared with littermate controls. This reduction was associated with lower levels of CitH3, a marker of neutrophil extracellular traps (NETs), in the harvested thrombi and improved IVC wall contraction and relaxation responses (assessed by myography). Mechanistically, thrombin-stimulated platelets from PKM2Plt-KO mice or ML265-pretreated platelets from WT mice showed reduced SNAP23 phosphorylation and diminished PF4 release (a marker of α-granule exocytosis). The releasate collected from thrombin-stimulated platelets was less effective at inducing NETosis, compared to respective controls. Utilizing ML265-pretreated human whole blood perfused over a tissue factor-coated surface at a venous shear rate, we found that the area covered by platelet-leukocyte aggregates was profoundly reduced compared to vehicle control. Consistent with murine data, human platelets pretreated with ML265 and stimulated with thrombin exhibited decreased PF4 release and generated releasates that were less potent in inducing NETosis. These findings reveal for the first time that targeting PKM2 genetically or pharmacologically reduces SNAP23-mediated α-granule exocytosis in platelets, platelet releasate-induced NETosis, and susceptibility to DVT.
Details
- Title: Subtitle
- Pyruvate kinase M2 promotes venous thrombosis by enhancing SNAP23-mediated platelet exocytosis and consequent NETosis
- Creators
- Manasa K Nayak - University of IowaGagan D Flora - University of IowaIvan Budnik - University of IowaTarun Barbhuyan - University of IowaRakesh B Patel - University of IowaMadankumar Ghatge - University of IowaMariia Kumskova - University of IowaAditi Jain - University of IowaNeelam Chauhan - University of IowaJitendra Kumar - University of IowaMegan Jewell - University of Colorado Anschutz Medical CampusSantosh Kumar - University of IowaSteven R Lentz - University of IowaKeith B Neeves - University of Colorado DenverAnil Chauhan - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Blood advances, Vol.10(1), pp.132-142
- DOI
- 10.1182/bloodadvances.2025017414
- PMID
- 41118613
- PMCID
- PMC12804119
- NLM abbreviation
- Blood Adv
- ISSN
- 2473-9529
- eISSN
- 2473-9537
- Publisher
- ELSEVIER
- Grant note
- National Institutes of Health, National Heart, Lung, and Blood Institute: R35HL139926, R01HL174460 American Heart Association career development award: 942168 National Institutes of Health, National Heart, Lung, and Blood Institute, United States: R01HL151984, R33HL141794 Department of Health and Human Services, Health Resources and Services Administration, United States: H30MC24049
The study is supported by grants from the National Institutes of Health, National Heart, Lung, and Blood Institute (R35HL139926 and R01HL174460) . M.K.N. is supported by the American Heart Association career development award (942168) . The laboratory of K.B.N. is supported by grants from National Institutes of Health, National Heart, Lung, and Blood Institute, United States (R01HL151984 and R33HL141794) and the Department of Health and Human Services, Health Resources and Services Administration, United States (H30MC24049, Hemophilia Treatment Centers) .
- Language
- English
- Electronic publication date
- 10/21/2025
- Date published
- 01/13/2026
- Academic Unit
- Hematology, Oncology, and Blood & Marrow Transplantation; Cardiovascular Medicine; Internal Medicine
- Record Identifier
- 9985017439202771
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