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321 Bleeding Diathesis in Mice Lacking JAK2 in Platelets

Program: Oral and Poster Abstracts
Type: Oral
Session: 301. Vascular Wall Biology, Endothelial Progenitor Cells, and Platelet Adhesion, Activation, and Biochemistry
Hematology Disease Topics & Pathways:
Diseases, Bleeding and Clotting, Hemostasis, Thrombosis, MPN, Polycythemia vera, thrombocythemia, Myeloid Malignancies
Sunday, December 6, 2020: 10:15 AM

Nathan Eaton, BS1,2*, Saravanan Subramaniam, PhD1*, Marie L Schulte, PhD1*, Caleb Drew, BS1*, David Jakab, BS1*, Sandra L Haberichter, PhD1,3,4, Hartmut Weiler, PhD1,5 and Herve Falet, PhD1,2

1Blood Research Institute, Versiti, Milwaukee, WI
2Department of Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee, WI
3Children's Wisconsin Research Institute, Children's Wisconsin, Milwaukee, WI
4Department of Pediatrics- Hematology/Oncology, Medical College of Wisconsin, Milwaukee, WI
5Department of Physiology, Medical College of Wisconsin, Milwaukee, WI

The tyrosine kinase JAK2 is a critical component of intracellular JAK/STAT cytokine signaling cascades that is prevalent in hematopoietic cells such as hematopoietic stem and progenitor cells (HSPCs), megakaryocytes (MKs), and platelets. Individuals expressing somatic activating JAK2 mutations such as JAK2V617F commonly develop myeloproliferative neoplasms (MPNs) associated with serious complications, including venous and arterial thrombosis, a leading cause of mortality. Here, we investigated the role of JAK2 in hemostasis and thrombosis using Jak2fl/fl Pf4-Cre (Jak2Plt–/–) mice specifically lacking JAK2 within the platelet lineage. Jak2Plt–/ mice developed severe thrombocytosis with a 5-fold increase in circulating platelet number, MK hyperplasia, and splenomegaly. Jak2Plt–/– platelets were of normal size and the expression of major membrane surface glycoproteins was indistinguishable from controls, except for the integrin β3, which was reduced by 20%. Despite the thrombocytosis, Jak2Plt–/– mice had a severe bleeding diathesis, as evidenced by: 1) prolonged tail bleeding time; 2) failure to occlude in a ferric chloride-induced carotid artery injury model; and 3) failure to form stable thrombi in a laser-induced cremaster muscle injury model. Jak2Plt–/– platelets spread poorly on immobilized collagen or on immobilized fibrinogen following GPVI stimulation with the collagen-related peptide (CRP). Jak2Plt–/– platelets had defective α-granule secretion and integrin αIIbβ3 activation following stimulation with CRP, but not thrombin, and showed aggregation defects with low-doses of CRP. Together, the data support a GPVI-specific impairment in platelets lacking JAK2, a notion that was supported by impaired intracellular signaling following GPVI stimulation, as assessed by protein tyrosine phosphorylation. Jak2Plt–/– platelets adhered poorly to type I collagen under arterial shear rates in whole blood. However, JAK2 deletion in platelets did not alter plasma von Willebrand factor (VWF) levels or botrocetin-mediated binding of plasma VWF to GPIbα. Together, the results underline a critical role for JAK2 in platelet GPVI signaling and hemostatic function, which likely contributes to venous and arterial thrombosis observed in patients with MPNs with the activating JAK2V617F mutation.

Disclosures: No relevant conflicts of interest to declare.

*signifies non-member of ASH