Dataset Information

MTOR regulates GPVI-mediated platelet activation.

ABSTRACT:

Background

Due to mTOR (mammalian/mechanistic target of rapamycin) gene-loss mice die during embryonic development, the role of mTOR in platelets has not been evaluated using gene knockout technology.

Methods

A mouse model with megakaryocyte/platelet-specific deletion of mTOR was established, and be used to evaluate the role of mTOR in platelet activation and thrombus formation.

Results

mTOR^-/- platelets were deficient in thrombus formation when grown on low-concentration collagen-coated surfaces; however, no deficiency in thrombus formation was observed when mTOR^-/- platelets were perfused on higher concentration collagen-coated surfaces. In FeCl₃-induced mouse mesenteric arteriole thrombosis models, wild-type (WT) and mTOR^-/- mice displayed significantly different responses to low-extent injury with respect to the ratio of occluded mice, especially within the first 40 min. Additionally, mTOR^-/- platelets displayed reduced aggregation and dense granule secretion (ATP release) in response to low doses of the glycoprotein VI (GPVI) agonist collagen related peptide (CRP) and the protease-activated receptor-4 (PAR4) agonist GYPGKF-NH₂; these deficiencies were overcame by stimulation with higher concentration agonists, suggesting dose dependence of the response. At low doses of GPVI or PAR agonist, the activation of α_IIbβ₃ in mTOR^-/- platelets was reduced. Moreover, stimulation of mTOR^-/- platelets with low-dose CRP attenuated the phosphorylation of S6K1, S6 and Akt Ser473, and increased the phosphorylation of PKCδ Thr505 and PKCε Ser729. Using isoform-specific inhibitors of PKCs (δ, ɛ, and α/β), we established that PKCδ/ɛ, and especially PKCδ but not PKCα/β or PKCθ, may be involved in low-dose GPVI-mediated/mTOR-dependent signaling.

Conclusion

These observations indicate that mTOR plays an important role in GPVI-dependent platelet activation and thrombus formation.

SUBMITTER: Wang L

PROVIDER: S-EPMC8111939 | biostudies-literature |

REPOSITORIES: biostudies-literature

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