Platelet proteomics reveals mechanism of platelet activation inhibition by AEE
Ontology highlight
ABSTRACT:
ORGANISM(S): Rattus Rattus
SUBMITTER: 
Yajun Yang
PROVIDER: PXD054906 | iProX | Wed Aug 14 00:00:00 BST 2024
REPOSITORIES: iProX
Similar Datasets
Project description:Platelets play a crucial role in cancer and thrombosis. The receptor-ligand repertoire mediating prostate cancer (PCa) cell-platelet interactions have not been fully elucidated. Microvilli emanating from the plasma membrane of PCa cell lines (RC77 T/E, MDA PCa 2b) directly contacted individual platelets and platelet aggregates, thereby stimulating calcium mobilization in platelets and promoting translocation of P-selectin and integrin aIIbb3 onto the platelet surface. Platelets in turn dose-dependently stimulated PCa cell invasion and apoptotic resistance. PCa cells were exceedingly sensitive to activation by platelets, occurring at platelet:PCa cell ratios ~20,000-fold lower than normally encountered in blood. Candidate platelet-PCa cell signaling axis partners responsible for these cellular events were identified by RNA-Seq and could be broadly divided into 4 categories: i) integrin-ligand, ii) EPH receptor-ephrin, iii) immune checkpoint receptor-ligand, and iv) miscellaneous receptor-ligand interactions. PCa cell-stimulated calcium mobilization was mediated by a fibronectin1 (FN1; PCa side)-aIIbb3 (platelet side) axis. Reciprocally, platelet-stimulated PCa cell invasion was facilitated by a CD55 (platelet side)-adhesion G-protein coupled receptor E5 (ADGRE5; PCa side) axis, with contribution from platelet cytokines CCL3L1 and IL32. Platelet-stimulated PCa cell apoptotic resistance relied on ephrin-EPH receptor and lysophosphatidic acid (LPA)-LPA receptor (LPAR) signaling. Of the participating partners, FN1 and LPAR3 overexpression was observed in PCa specimens compared to normal prostate, while high expression of CCR1 (CCL3L1 receptor), EPHA1 and LPAR5 in PCa was associated with poor patient survival. These findings emphasize that non-overlapping receptor-ligand pairs participate in oncogenesis and thrombosis, highlighting the complexity of any contemplated clinical intervention strategy.
2022-11-05 | GSE216942 | GEO
Project description:Acute myocardial infarction (AMI) is primarily due to coronary atherosclerotic plaque rupture and subsequent thrombus formation. Platelets play a key role in the genesis and progression of both atherosclerosis and thrombosis. Since platelets are anuclear cells that inherit their mRNA from megakaryocyte precursors and maintain it unchanged during their life span, gene expression (GE) profiling at the time of an AMI provides information concerning the platelet GE preceding the coronary event. In ST-segment elevation myocardial infarction (STEMI), a gene-by-gene analysis of the platelet GE identified five differentially expressed genes (DEGs): FKBP5, S100P, SAMSN1, CLEC4E and S100A12. The logistic regression model used to combine the GE in a STEMI vs healthy donors score showed an AUC of 0.95. The same five DEGs were externally validated using platelet GE data from patients with coronary atherosclerosis but without thrombosis. Early signals of an imminent AMI are likely to be found by platelet GE profiling before the infarction occurs.
2019-12-31 | GSE109048 | GEO
Project description:Renowned for their role in hemostasis and thrombosis, platelets are increasingly recognized for their involvement in innate immunity, immunothrombosis and inflammatory diseases. Platelets, heterogeneous in size and molecular content, express a wide range of receptors enabling versatile activation endpoints relevant for immune functions. Upon activation, platelet release extracellular vesicles (PEVs), whose formation and molecular cargo exhibit remarkable tunability based on receptor-mediated activation or environmental cues. This study aimed to compare the immune modulatory roles of PEVs released through activation by specific platelet receptors, GPVI, CLEC-2, and thrombin-collagen (TC). Functional assays in vivo in zebrafish and using an in vitro human macrophage models highlighted distinct immune-inflammatory responses triggered by PEVs. Surprisingly, despite extensive characterization using omics analyses and basic EV particle characterization, the differences in the protein and miRNA cargo and the EV physicochemical properties between the PEV types were subtle and insufficient to precisely define their mechanism of action to distinguish a distinct functional profile. Moreover, the constitutively formed PEVs used as controls displayed a disparate activation profile from the receptor-induced PEVs.
2024-09-19 | PXD047323 | Pride
Project description:For the anucleate platelet it has been unclear how well platelet transcriptomes correlate among different donors or across different RNA profiling platforms, and what the transcriptomes’ relationship is with the platelet proteome. We generated RNA-seq pro-files of the long RNA transcriptomes from the platelets of 10 healthy young males (5 white and 5 black) with median age of 24.5 years, no notable clinical history, and no pre-vious history of thrombosis or bleeding. We also profiled the subjects’ messenger RNAs using the Affymetrix microarray gene expression system. We found that the abundance of platelet mRNA transcripts was highly correlated across the 10 individuals, inde-pendently of race and of the employed technology. Our RNA-seq data also showed that these high inter-individual correlations extend beyond mRNAs to several categories of non-coding RNAs. Pseudogenes represented a notable exception to this by exhibiting a clear difference in expression by race. Comparison of our mRNA signatures with the only publicly available quantitative platelet proteome data showed that most (87.5%) identified platelet proteins had a detectable corresponding mRNA. However, a high number of mRNAs that were present in the transcriptomes of all 10 individuals had no representa-tion in the proteome. The Spearman correlation of the relative abundances for those platelet genes that were represented by both an mRNA and a protein showed a weak (~0.3) yet statistically significant (P=5.0E-16) connection. Further analysis of the overlap-ping and non-overlapping platelet mRNAs and proteins identified gene groups corre-sponding to distinct cellular processes, a finding that provides novel insights for platelet biology. This represents the Affymetrix GeneChip component of the study only 10 Resting Human Platelet samples
2015-01-20 | E-GEOD-50858 | biostudies-arrayexpress
Project description:The GPIbα cytoplasmic tail peptide MPαC was used to study the role of GPIbα in platelet activation.
2024-04-02 | PXD051165 |
Project description:Platelets engage cues of pending endothelial dysfunction through coordinated adhesion, secretion and aggregation responses. These rapid changes in platelet phenotype are orchestrated by intracellular mechanisms that remain systematically undefined. This study develops a TMT-SPS-MS3 phosphoproteomics workflow to detail ~3,800 significant protein phosphorylation events associated with the progression of ITAM-mediated activation programs initiated by the platelet collagen receptor GPVI. With literature-guided causal inference tools, >200 site-specific signaling relations are mapped from phosphoproteomics data among key GPVI effectors (i.e., Syk, PLC gamma 2, PKC delta) and less characterized targets, including several Ras/MAPK axis proteins (i.e., KSR1). Networks highly regulated in GPVI/ITAM signaling out of context of curated knowledge are also highlighted, including Rab GTPase systems. In addition to serving as a model for generating and testing hypotheses from omics datasets, this study puts forth a means to identify hemostatic effectors and biomarkers relevant to thrombosis, vascular inflammation and other platelet-associated disease states.
2020-11-20 | PXD017167 | Pride
Project description:Platelets are key players in hemostasis and thrombosis. In inflammatory conditions, platelets are hyperreactive and patients have an overal increased thrombotic risk. Our previous research demonstrated that TNFα significantly increases mitochondrial mass and platelet reactivity, emphasizing the crucial role of TNFα in modulating platelet metabolism and function.We hypothesized that TNFα disrupts platelet metabolism and function by inhibiting autophagy and mitophagy, which are critical processes for preserving mitochondrial mass, integrity, and energy production. We investigated the function, metabolism, and autophagy of human and mouse platelets in myeloproliferative neoplasms, rheumatoid arthritis, and aseptic inflammation. Our analysis reveled a blocked
2025-03-31 | GSE282993 | GEO
Project description:Platelet activators stimulate post-translational modification of signalling proteins to change their activity or their molecular interactions leading to signal propagation. One covalent modification is attachment of the small protein ubiquitin to lysine residues in target proteins. Modification by ubiquitin can either target proteins for degradation by the proteasome or act as a scaffold for other proteins. Pharmacological inhibition of deubiquitylases or the proteasome inhibits platelet activation by collagen, demonstrating a role for ubiquitylation, but relatively few substrates for ubiquitin have been identified and the molecular basis of inhibition is not established. Here we report the ubiquitome of human platelets and changes in ubiquitylated proteins following stimulation by collagen related peptide (CRP-XL). We identified 1634 ubiquitylated peptides derived from 691 proteins, revealing extensive ubiquitylation in resting platelets. 925 of these peptides show an increase of more than 2-fold following stimulation with CRP-XL. Multiple sites of ubiquitylation were identified on a number of proteins including Syk, filamin and integrins. Adhesion and spreading on fibrinogen mediated by the major platelet integrin IIb3 is blocked by inhibition of deubiquitylases. This work reveals extensive protein ubiquitylation during activation of human platelets and opens the possibility of novel therapeutic interventions targeting the ubiquitin machinery.
2019-01-07 | PXD009072 | Pride
Project description:MicroRNAs (miRNAs) regulate cell physiology by altering protein expression, but the biology of platelet miRNAs is largely unexplored. We tested whether platelet miRNA levels were associated with platelet reactivity by genome-wide profiling using platelet RNA from 19 healthy subjects. We found that human platelets express 284 miRNAs. Unsupervised hierarchical clustering of miRNA profiles resulted in 2 groups of subjects that appeared to cluster by platelet aggregation phenotypes. Seventy-four miRNAs were differentially expressed (DE) between subjects grouped according to platelet aggregation to epinephrine, a subset of which predicted the platelet reactivity response. Using whole genome mRNA expression data on these same subjects, we computationally generated a high-priority list of miRNA-mRNA pairs in which the DE platelet miRNAs had binding sites in 3'UTRs of DE mRNAs, and the levels were negatively correlated. Three miRNA-mRNA pairs (miR-200b:PRKAR2B, miR-495:KLHL5 and miR-107:CLOCK) were selected from this list and all 3 miRNAs knocked down protein expression from the target mRNA. Reduced activation from platelets lacking PRKAR2B supported these findings. In summary, (1) platelet miRNAs are able to repress expression of platelet proteins, (2) miRNA profiles are associated with and may predict platelet reactivity, and (3) bioinformatic approaches can successfully identify functional miRNAs in platelets. Total RNA from the platelets of 19 donors was harvested and labeled with Hy3. Reference RNA (a pool of all samples) was labeled with Hy5. This submission represents the miRNA expression component of the study.
2011-09-26 | E-GEOD-27917 | biostudies-arrayexpress
Project description:MTH1 knockout mice were utilized to evaluate its role in platelet mitochondrial function and thrombosis
2023-04-14 | PXD041528 |