Expression data from human platelets after purification
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE193664 | GEO | 2022/03/31
REPOSITORIES: GEO
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Project description:Abstract. The role of platelets in hemostasis and thrombosis is clearly established; however, the mechanisms by which platelets mediate inflammatory and immune pathways are less well understood. Platelets interact and modulate the function of blood and vascular cells by releasing bioactive molecules. Although the platelet is anucleate, it contains transcripts that may mirror disease. Platelet mRNA is only associated with low-level protein translation, however, platelets have a unique membrane structure allowing for the passage of small molecules, leading to the possibility that its’ cytoplasmic RNA may be passed to nucleated cells. To examine this question, platelet-like particles with labeled RNA were co-cultured with vascular cells. Co-culture of platelet-like particles with activated THP-1, monocytic, and endothelial cells led to visual and functional RNA transfer. Post-transfer microarray gene expression analysis of THP-1 cells showed an increase in HBG1/HBG2 and HBA1/HBA2 expression which was directly related to the transfer. Infusion of wild-type platelets into a TLR2 deficient mouse model established in vivo confirmation of select platelet RNA transfer to leukocytes. By specifically transferring green fluorescent protein, it was also observed that external RNA was functional in the recipient cells. The observation that platelets possess the capacity to transfer cytosolic RNA suggests a new function for platelets in the regulation of vascular homeostasis. THP-1, treated with Pam3CSK4, were co-cultured with platelet-like particles for 12 hours and 24 hours then collected and processed for RNA isolation. We also included as control THP-1 untreated and THP-1 treated with Pam3CSK4. A total of 4 samples were analyzed by microarray for gene expression.
2012-05-17 | E-GEOD-38027 | biostudies-arrayexpress
Project description:Abstract. The role of platelets in hemostasis and thrombosis is clearly established; however, the mechanisms by which platelets mediate inflammatory and immune pathways are less well understood. Platelets interact and modulate the function of blood and vascular cells by releasing bioactive molecules. Although the platelet is anucleate, it contains transcripts that may mirror disease. Platelet mRNA is only associated with low-level protein translation, however, platelets have a unique membrane structure allowing for the passage of small molecules, leading to the possibility that its’ cytoplasmic RNA may be passed to nucleated cells. To examine this question, platelet-like particles with labeled RNA were co-cultured with vascular cells. Co-culture of platelet-like particles with activated THP-1, monocytic, and endothelial cells led to visual and functional RNA transfer. Post-transfer microarray gene expression analysis of THP-1 cells showed an increase in HBG1/HBG2 and HBA1/HBA2 expression which was directly related to the transfer. Infusion of wild-type platelets into a TLR2 deficient mouse model established in vivo confirmation of select platelet RNA transfer to leukocytes. By specifically transferring green fluorescent protein, it was also observed that external RNA was functional in the recipient cells. The observation that platelets possess the capacity to transfer cytosolic RNA suggests a new function for platelets in the regulation of vascular homeostasis.
2012-05-18 | GSE38027 | GEO
Project description:Beyond a function in hemostasis and thrombosis, platelets can regulate innate and adaptive immune responses. Hyperactive platelets are frequently associated with multiple human autoimmune diseases, yet their pathogenic functions in these diseases have not been fully established. Emerging studies show an essential function of the phosphatase and tensin homolog (PTEN) in maintenance of immune homeostasis. Here, we show that mice with platelet-specific deletion of Pten, develop age-related lymphoproliferative diseases and humoral autoimmunity not seen in wildtype animals. Platelet-specific Pten-deficient mice have aberrant T cell activation, excessive T follicular helper (Tfh) cell responses and accumulation of platelet aggregates in lymph nodes. Transferred Pten-deficient platelets are able to infiltrate into the peripheral lymphoid tissues and form more aggregates. Moreover, Pten-deficient platelets are hyperactive and overproduce multiple Tfh-promoting cytokines via activation of the PDK1/mTORC2-AKT-SNAP23 pathway. Pten-deficient platelets show enhanced interaction with CD4+ T cells and promote conversion of CD4+ T cells into Tfh cells. Our results implicate PTEN in platelet-mediated immune homeostasis, and provide evidence that hyperactive platelets function as an important mediator in autoimmune diseases using mouse models.
2021-01-01 | GSE143698 | GEO
Project description:Platelets are anucleate cytoplasmic fragments that lack genomic DNA, but continue to synthesize protein using a pool of mRNAs, ribosomes, and regulatory small RNAs inherited from the precursor megakaryocyte (MK). The regulatory processes that shape the platelet transcriptome and the full scope of platelet translation have remained elusive. Using RNA-Seq and ribosome profiling of primary human platelets, we show the platelet transcriptome encompasses a subset of transcripts detected by RNA-Seq analysis of in vitro derived MK cells and these platelet-enriched transcripts are broadly occupied by ribosomes. We use RNA sequencing of synchronized populations of in vitro derived platelet-like particles (PLPs) to show that mRNA decay strongly shapes the nascent platelet transcriptome. Our data suggests that the decay of platelet mRNAs is slowed by the natural loss of the mRNA surveillance and ribosome rescue factor Pelota (PELO).
2017-02-02 | GSE94384 | GEO
Project description:Platelets are anucleated blood cells that are produced by their progenitor cell, the megakaryocyte (MK). Platelets are centrally positioned in hemostasis and thrombosis and, according to the recent findings, play key roles in innate immunity, inflammation, atherogenesis, and cancer metastasis. The quantitative and qualitative properties of platelets are crucial determinants of their hemostatic function. While the regulatory mechanisms of platelet number and size have been studied separately, critical questions remain regarding the interplay of platelet number and size in hemostasis. In this study, using a mouse model of irradiation (IR)-induced thrombocytopenia, we show that mature MKs (mMKs) are resistant to IR and maintain the nadir platelet levels post IR. To identify the phenotype switching and function variation in BM MKs post IR, we performed microarray expression profiling of MKs from the bone marrow of mice at 1- or 3-days post IR and control mice (Ctrl).
2023-12-27 | GSE222512 | GEO
Project description:Platelet activation is the key event triggering thrombus formation in physiological and pathological conditions, such as acute coronary syndromes. Current therapies using antiaggregants still fail to prevent thrombotic coronary events in a significant number of patients, indicating that the mechanisms modulating platelet response during activation need to be clarified. The evidence that platelets are capable of de novo protein synthesis in response to stimuli raised the issue of how the activity of megakaryocyte-derived mRNAs is regulated in these anucleate cell fragments. We applied a combined multi-omics approach to investigate this phenomenon in platelets from healthy donors activated in vitro with Collagen or Thrombin Receptor Activating Peptide. Combining HiRIEF LC-MS to transcriptome analysis by RNA-Seq allowed platelet proteome characterization at deep coverage, revealing a significant effect of either stimulus on proteome composition. In silico intron retention analysis was then applied to search for splicing events induced by platelet activation, coupled to unbiased proteogenomics, to correlate intron retention in resting platelets to intron removal by RNA splicing during activation. This allowed identification of a set of transcripts, specifically involved in platelet shape changes, showing reduced intron retention and high peptide representation at exon-exon junctions in activated vs resting platelets. These results indicate that RNA splicing events takes place in platelets during activation and that pre-mRNA maturation of specific transcripts is part of the activation cascade and could therefore provide novel molecular markers of platelet activation status in acute coronary syndromes and other pathological conditions.
2018-01-15 | E-MTAB-6148 | biostudies-arrayexpress
Project description:Platelets are involved in relevant physiological and pathophysiological processes such as hemostasis and tumor progression. These anucleate blood particles exert their functions essentially by releasing effector molecules such as proteins from intracellular granules and by generating and secreting specific lipids. In order to contribute to a systematic proteomics and eicosadomics analysis of platelets activated by calcium mobilization, we have investigated proteins and eicosanoids in lysates and secretomes of platelets activated with ionomycin. To this end, self-established shotgun proteomics and shotgun lipidomics platforms were used. Applying an FDR of less than 0.01 at both, peptide and protein level, a total of 2543 protein groups was identified. Several regulatory effects concerning proteins were determined, including protein secretion, degradation, as well as protein synthesis. Incorporation of 35S-methionine in platelet proteins determined by 2D-PAGE confirmed de-novo protein synthesis. Moreover, we were able to determine a complex set of eicosanoids synthesized by activated platelets, including 15S-HETE, prostaglandin E2 and pro-survival factors such as 12-HETE, as well as eicosanoids previously undescribed to have platelet functions, such as 9-HETE and 11-HETE. Proteomics data complemented the findings from lipidomics experiments and supported the generation of a map indicating pathways leading to eicosanoid production in platelets upon calcium mobilization.
2022-03-01 | PXD005054 | Pride
Project description:Platelets contain abundant miRNAs, however, the biogenesis pathway of miRNAs in anucleate platelets is unclear. Platelet-rich plasma was diluted in washing buffer and the platelet suspension was centrifuged to isolated pure platelets.Finally, platelets were recovered in suspension buffer at the concentration of 4–5×10^8 platelets/ml. Aliquots of the platelet suspensions were activated in the presence of 2.5 mM CaCl2 with 0ng/ml or 1 ng/ml thrombopoietin (TPO)
2014-02-04 | E-GEOD-51453 | biostudies-arrayexpress
Project description:Platelets are anucleate blood cells that contain mitochondria, that regulate blood clotting in response to injury. Mitochondria contain their own gene expression machinery which relies on nuclear encoded factors for the biogenesis of the oxidative phosphorylation (OXPHOS) system to produce energy, ions and metabolites required for thrombosis. The autonomy of the mitochondrial gene expression machinery from the nucleus is not known and platelets provide a valuable model to understand the importance of mitochondrial gene expression in anucleate cells. We generated three different platelet-specific conditional knockout mouse models, each lacking a gene that is essential for mitochondrial gene expression at the level of RNA processing, stability and translation (Elac2Pf4∆/Pf4∆, Ptcd1Pf4∆/Pf4 and Mtif3Pf4∆/Pf4∆). Loss of ELAC2, PTCD1 or MTIF3, leads to increased megakaryocyte ploidy, elevated circulating levels of reticulated platelets, thrombocytopenia and consequent extended bleeding time. Loss of ELAC2, PTCD1 and MTIF3 reduced agonist induced platelet activation. Transcriptomic and proteomic analyses showed that mitochondrial gene expression and protein synthesis facilitate fibrinolysis, haemostasis and blood coagulation in response to injury.
2023-10-03 | GSE206543 | GEO
Project description:Ex vivo assays of platelet function critically inform mechanistic and clinical studies of hemostasis and thrombosis, where effects of divergent blood processing methods on platelet composition are apparent but remain unspecified. Parallel blood samples were collected from healthy human donors into sodium citrate, acid citrate dextrose, EDTA and heparin, and processed over an extended time course for physiological and biochemical experiments, including platelet proteome quantification with multiplexed tandem mass tag (TMT) labeling and high-resolution Tribrid mass spectrometry (MS). We evaluated how different blood anticoagulation options and processing times affect platelet protein content ex vivo. Following platelet isolation, TMT-MS quantified 3,358 proteins amongst all prepared platelet samples. Altogether, >400 proteins were differentially abundant in platelets isolated from blood processed at 24 h vs. 1 h post-phlebotomy, including sets of proteins pertinent to membrane trafficking and exocytosis.
2023-01-17 | PXD030225 | Pride