Document Type
Article
Publication Date
3-26-2024
Abstract
Mechanisms of proteostasis in anucleate circulating platelets are unknown and may regulate platelet function. We investigated the hypothesis that plasma-borne growth factors/hormones (GFHs) maintain constitutive translation in circulating platelets to facilitate reactivity. Bio-orthogonal noncanonical amino acid tagging (BONCAT) coupled with liquid chromatography-tandem mass spectrometry analysis revealed constitutive translation of a broad-spectrum translatome in human platelets dependent upon plasma or GFH exposure, and in murine circulation. Freshly isolated platelets from plasma showed homeostatic activation of translation-initiation signaling pathways: phosphorylation of p38/ERK upstream kinases, essential intermediate MNK1/2, and effectors eIF4E/4E-BP1. Plasma starvation led to loss of pathway phosphorylation, but it was fully restored with 5-minute stimulation by plasma or GFHs. Cycloheximide or puromycin infusion suppressed ex vivo platelet GpIIb/IIIa activation and P-selectin exposure with low thrombin concentrations and low-to-saturating concentrations of adenosine 5'-diphosphate (ADP) or thromboxane analog but not convulxin. ADP-induced thromboxane generation was blunted by translation inhibition, and secondary-wave aggregation was inhibited in a thromboxane-dependent manner. Intravenously administered puromycin reduced injury-induced clot size in cremaster muscle arterioles, and delayed primary hemostasis after tail tip amputation but did not delay neither final hemostasis after subsequent rebleeds, nor final hemostasis after jugular vein puncture. In contrast, these mice were protected from injury-induced arterial thrombosis and thrombin-induced pulmonary thromboembolism (PE), and adoptive transfer of translation-inhibited platelets into untreated mice inhibited arterial thrombosis and PE. Thus, constitutive plasma GFH-driven translation regulates platelet G protein-coupled receptor reactivity to balance hemostasis and thrombotic potential.
Recommended Citation
Wurtzel, Jeremy G.T.; Lazar, Sophia; Askari, Shayan; Zhao, Xuefei; Severa, Jenna; Ayombil, Francis; Michael, James V.; Camire, Rodney M.; McKenzie, Steven E.; Stalker, Timothy J.; Ma, Peisong; and Goldfinger, Lawrence E., "Plasma Growth Factors Maintain Constitutive Translation in Platelets to Regulate Reactivity and Thrombotic Potential" (2024). Cardeza Foundation for Hematologic Research. Paper 80.
https://jdc.jefferson.edu/cardeza_foundation/80
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.
Supplemental Table 2.xlsx (21 kB)
Supplemental Table 3.xlsx (1874 kB)
Supplemental Methods, Figures, Legends for Tables, and References.pdf (1107 kB)
Language
English
Comments
This article is the author's final published version in Blood Advances, Volume 8, Issue 6, March 2024, Pages 1550 - 1566.
The published version is available at https://doi.org/10.1182/bloodadvances.2023011734.
Copyright © 2024 by The American Society of Hematology.