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Functionally-relevant splicing of resident pre-mRNAs in anucleate platelets upon activation by physiological stimuli

ABSTRACT: Functionally-relevant splicing of resident pre-mRNAs in anucleate platelets upon activation by physiological stimuli

PROVIDER: PRJEB23127 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR2179070_1.fastq.gz	Fastqsanger.gz
	ERR2179070_2.fastq.gz	Fastqsanger.gz
	ERR2179071_1.fastq.gz	Fastqsanger.gz
	ERR2179071_2.fastq.gz	Fastqsanger.gz
	ERR2179072_1.fastq.gz	Fastqsanger.gz

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Functionally-relevant splicing of resident pre-mRNAs in anucleate platelets upon activation by physiological stimuli

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.

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