GRK6 Regulates the Hemostatic Response to Injury through Its Rate-Limiting Effects on GPCR Signaling in Platelets

Chen, Xi; Gupta, Shuchi; Cooper, Matthew; DeHelian, Daniel; Zhao, Xuefei; Brass, Lawrence F.; Naik, Ulhas P.; Naik, Meghna U.; Benovic, Jeffrey L.; Ma, Peisong

doi:10.1182/blood-2019-129175

Cited by 4 publications

(25 citation statements)

References 39 publications

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“…Messenger RNA and predicted protein copy number analysis in human and mouse platelets have identified the presence of GRK expression [ 21 , 22 , 23 ]. Further study using immunoblotting has confirmed the expression of specific GRKs (GRK2, GRK5, and GRK6) in platelets [ 24 , 25 , 26 ]. In human platelets, GRK2, GRK5, and GRK6 are all expressed in the range of 1000–2000 copies per platelet, with GRK6 having the highest expression level.…”

Section: Grks In Plateletsmentioning

confidence: 97%

“…In both human and mouse platelets, GRK6 is the predominant form of GRKs. It impacts platelet activation through targeting of the PARs and P2Y 12 receptors ( Figure 1 A) [ 25 ]. Human platelets express thrombin receptors, PAR1 and PAR4, but mouse platelets express a PAR3/PAR4 complex [ 37 , 38 ].…”

Section: The Role Of Grks and Arrestins During Platelet Activationmentioning

confidence: 99%

“…Using human megakaryoblastic cells (MEG-01), it has been shown that there is an increase in Ca 2+ response to PAR1 agonist in GRK6 -/- cells, but no change in Ca 2+ response to PAR4 agonist. This suggests that GRK6 regulates PAR1, but not PAR4, receptor-mediated signaling in human platelets [ 25 ]. Using CRISPR-Cas9 genome editing, we generated a GRK6 knockout mouse mutant line.…”

Section: The Role Of Grks and Arrestins During Platelet Activationmentioning

confidence: 99%

“…Using CRISPR-Cas9 genome editing, we generated a GRK6 knockout mouse mutant line. We have shown that loss of GRK6 in mouse platelets increases both PAR4 and P2Y 12 -dependent signaling [ 25 ]. Notably, deletion of GRK6 has an effect on mouse PAR4-dependent signaling but not human PAR4-dependent signaling.…”

Section: The Role Of Grks and Arrestins During Platelet Activationmentioning

confidence: 99%

“…This is because GRK6 mainly phosphorylates the C-terminus serine/threonine residues of GPCRs, such as TPβ, but cannot sufficiently phosphorylate the relatively shorter C-terminus of TPα. Therefore, GRK6 might not trigger TPα internalization and subsequent TPα-mediated signaling in platelets [ 25 , 26 ].…”

Section: The Role Of Grks and Arrestins During Platelet Activationmentioning

confidence: 99%

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The Roles of GRKs in Hemostasis and Thrombosis

Chen

Zhao

Cooper

et al. 2020

IJMS

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Along with cancer, cardiovascular and cerebrovascular diseases remain by far the most common causes of death. Heart attacks and strokes are diseases in which platelets play a role, through activation on ruptured plaques and subsequent thrombus formation. Most platelet agonists activate platelets via G protein-coupled receptors (GPCRs), which make these receptors ideal targets for many antiplatelet drugs. However, little is known about the mechanisms that provide feedback regulation on GPCRs to limit platelet activation. Emerging evidence from our group and others strongly suggests that GPCR kinases (GRKs) are critical negative regulators during platelet activation and thrombus formation. In this review, we will summarize recent findings on the role of GRKs in platelet biology and how one specific GRK, GRK6, regulates the hemostatic response to vascular injury. Furthermore, we will discuss the potential role of GRKs in thrombotic disorders, such as thrombotic events in COVID-19 patients. Studies on the function of GRKs during platelet activation and thrombus formation have just recently begun, and a better understanding of the role of GRKs in hemostasis and thrombosis will provide a fruitful avenue for understanding the hemostatic response to injury. It may also lead to new therapeutic options for the treatment of thrombotic and cardiovascular disorders.

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Section: Grks In Plateletsmentioning

confidence: 97%

Section: The Role Of Grks and Arrestins During Platelet Activationmentioning

confidence: 99%

Section: The Role Of Grks and Arrestins During Platelet Activationmentioning

confidence: 99%

Section: The Role Of Grks and Arrestins During Platelet Activationmentioning

confidence: 99%

Section: The Role Of Grks and Arrestins During Platelet Activationmentioning

confidence: 99%

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The Roles of GRKs in Hemostasis and Thrombosis

Chen

Zhao

Cooper

et al. 2020

IJMS

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Tripeptide Hyp‐Asp‐Gly from collagen peptides inhibited platelet activation via regulation of PI3K/Akt‐MAPK/ERK1/2 signaling pathway

Song

Yang

Лі

2022

Journal of Food Science

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Platelet activation is involved in cardiovascular thrombosis. Our previous study demonstrated that oral administration of collagen peptides (CPs) inhibited platelet activation, but the mechanism of action of CPs remained to be elucidated. As a continued effort, the objective of this study was to identify the active ingredient of CPs and clarify its molecular mechanism. Simulated absorbate of CPs was prepared by simulated gastrointestinal digestion and intestinal absorption system, and then separated by C18 column. The fraction with the highest antiplatelet activity was subjected to NanoUPLC-ESI-MS/MS for peptide sequencing. Novel tripeptide Hyp-Asp-Gly (ODG) was identified. It had a broad-spectrum inhibition of platelet activation induced by collagen, thrombin, and adenosine diphosphate (ADP). ODG could survive simulated gastrointestinal digestion and be absorbed intact. Furthermore, it showed good stability in plasma. ODG had no significant effect on the PLC-PKC-Ca 2+ pathway, but it inhibited the PI3K/Akt-MAPK/ERK1/2 signaling. At a dosage of 200 µmol/kg body weight, ODG had an in vivo anti-thrombosis activity without bleeding risk.The present study provides one of the mechanisms of action of CPs and highlights its potential use as a functional component to combat cardiovascular thrombosis.

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Human and mouse PAR4 are functionally distinct receptors: Studies in novel humanized mice

Renna

Michael

Kong

et al. 2022

Journal of Thrombosis and Haemostasis

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Background Human and mouse platelets both express protease‐activated receptor (PAR) 4 but sequence alignment reveals differences in several functional domains. These differences may result in functional disparities between the receptors which make it difficult to translate PAR4 studies using mice to human platelet physiology. Objectives To generate transgenic mice that express human, but not mouse, PAR4 and directly compare human and mouse PAR4 function in the same platelet environment. Methods Transgenic mice were made using a genomic clone of the F2RL3 gene (encoding PAR4) and backcrossed with Par4 KO mice. For certain experiments, mice were bred with GRK6 KO mice. Tail bleeding time and platelet function in response to PAR4‐activating peptide were assessed. Results Human F2RL3 was successfully integrated into the mouse genome, transgenic mice were crossed to the mPar4 KO background (PAR4 tg/KO), and PAR4 was functionally expressed on platelets. Compared to WT, PAR4 tg/KO mice exhibited shortened tail bleeding time and their platelets were more responsive to PAR4‐AP as assessed by α‐granule release and integrin activation. The opposite was observed with thrombin. Knocking out GRK6 had no effect on human PAR4‐expressing platelets, unlike mouse Par4‐expressing platelets. PAR4 tg/KO platelets exhibited greater Ca2+ area under the curve and more robust extracellular vesicle release than WT stimulated with PAR4‐AP. Conclusion These data suggest that (1) human PAR4‐ and mouse Par4‐mediated signaling are different and (2) the feedback regulation mechanisms of human and mouse PAR4 are different. These functional differences are important to consider when interpreting PAR4 studies done with mice.

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GRK6 Regulates the Hemostatic Response to Injury through Its Rate-Limiting Effects on GPCR Signaling in Platelets

Cited by 4 publications

References 39 publications

The Roles of GRKs in Hemostasis and Thrombosis

The Roles of GRKs in Hemostasis and Thrombosis

Tripeptide Hyp‐Asp‐Gly from collagen peptides inhibited platelet activation via regulation of PI3K/Akt‐MAPK/ERK1/2 signaling pathway

Human and mouse PAR4 are functionally distinct receptors: Studies in novel humanized mice

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