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Small non-coding RNAs in extracellular vesicles derived from equine synovial fluid and plasma in a model of equine osteoarthritis

ABSTRACT: Small non-coding RNAs in extracellular vesicles derived from equine synovial fluid and plasma in a model of equine osteoarthritis

PROVIDER: PRJNA802211 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17835800.fastq.gz	Fastqsanger.gz
	SRR17835801.fastq.gz	Fastqsanger.gz
	SRR17835802.fastq.gz	Fastqsanger.gz
	SRR17835803.fastq.gz	Fastqsanger.gz
	SRR17835804.fastq.gz	Fastqsanger.gz

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Small non-coding RNAs in extracellular vesicles derived from equine synovial fluid and plasma in a model of equine osteoarthritis

Project description:We studied extracellular vesicles (EVs) from plasma and synovial fluid in an in vivo model of equine osteoarthritis by investigating longitudinal samples. EVs were isolated using size exclusion chromatography from plasma and synovial fluid of four horses subjected to an osteochondral fragment model of osteoarthritis at 0, 10, 35, 42, 49, 56, 63 days with relevant controls.EV RNA was extracted and subject to small RNA sequencing on an Illumina NovaSeq SP using 100bp, single end reads. After mapping against EquCab.3.0 and miRBase v22.1 differential expression analysis was undertaken with edgeRv3.28 using a quasi-likelihood negative binomial generalized log-linear model. We identified a panel of altered small non-coding RNAs.

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Multiomic temporal landscape of plasma and synovial fluid derived extracellular vesicles using an experimental model of equine osteoarthritis

Project description:Osteoarthritis (OA) is the most common equine joint disease characterised by cartilage degradation and changes to other joint tissues severely effecting welfare and performance leading to early retirement. It results in substantial morbidity and mortality, but our understanding of the pathophysiological mechanisms involved is limited. There are no disease modifying therapeutics available, with OA medication offering symptomatic relief only. Early diagnosis is thus important, as substantial joint pathology is usually present at the time of clinical diagnosis. Thus, biomarkers of early-stage OA are actively sought, and among these, extracellular vesicles (EVs) are emerging as potential candidates. Here we use a SWATH MS approach to profile the protein cargo of EVs derived from equine plasma and synovial fluid.

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Investigation of MicroRNA Biomarkers in Equine Distal Interphalangeal Joint Osteoarthritis

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Small non-coding RNAs in early equine osteoarthritis synovial fluid

Project description:Small non-coding RNAs in early equine osteoarthritis synovial fluid

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miRNA sequencing of synovial fluid derived extracellular vesicles between normal and osteoarthritis (OA) patients

Project description:To determine whether extracellular vesicles were also present in human disease, we isolated and evaluated extracellular vesicles from synovial fluid from normal and OA patients at relatively advanced ages (70 - 80 years). Here we report the altered expression of synovial fluid extracellular vesicles-derived miRNAs in osteoarthritis compared to normal patients with age-relate degeneration to investigate potential biomarkers for OA diagnosis. The OA donors had clinical evidence of OA based on pain that led to total joint arthroplasty.

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Characterization of extracellular vesicles in plasma and synovial fluid of horses with naturally occurring post-traumatic osteoarthritis: An exploratory study to identify transcriptomic profiles.

Project description:Objective: The objective of this study was to characterize extracellular vesicles (EVs) in plasma and synovial fluid obtained from horses with and without naturally occurring post-traumatic osteoarthritis (PTOA). Animals (Samples): EVs were isolated from plasma and synovial fluid from horses with (n = 6) and without (n = 6) PTOA. Methods: Plasma and synovial fluid EVs were characterized with respect to quantity, size, and surface markers. Small RNA sequencing was performed and differentially expressed miRNAs underwent bioinformatic analysis to identify putative targets and to explore potential associations with specific biological processes. Results: Plasma and synovial fluid samples from horses with PTOA had a significantly higher proportion of exosomes and a lower proportion of microvesicles compared to horses without PTOA. Small RNA sequencing revealed several differentially expressed miRNAs including miR-144, miR-219-3p, and miR-199a-3p in plasma and miR-199a-3p, miR-214, and miR-9094 in synovial fluid EVs. Bioinformatics analysis of the differentially expressed miRNAs highlighted their potential role in fibrosis, differentiation of chondrocytes, apoptosis, and inflammation pathways in PTOA. Clinical Relevance: We have identified dynamic molecular changes in small non-coding signatures of plasma and synovial fluid EVs in horses with naturally occurring PTOA. These findings could serve to identify promising biomarkers in the pathogenesis of PTOA, to facilitate the development of targeted therapies, and to aid in establishing appropriate translational models of PTOA.

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Equine synovial fluid

Project description:The aim of the study was to characterise the protein complement of synovial fluid (SF) in health and osteoarthritis (OA) using liquid chromatography mass spectrometry (LC-MS/MS) following peptide-based depletion of high abundance proteins. SF was used from nine normal and nine OA Thoroughbred horses. Samples were analysed with LC-MS/MS using a NanoAcquity™ LC coupled to a LTQ Orbitrap Velos. In order to enrich the lower-abundance protein fractions protein equalisation was first undertaken using ProteoMiner™. Progenesis-QI™ LC-MS software was used for label-free quantification. In addition immunohistochemistry, western blotting and mRNA expression analysis was undertaken on selected joint tissues.The number of protein identifications was increased by 33% in the Proteominer™ treated SF compared to undepleted SF. A total of 754 proteins were identified in SF. A subset of 10 proteins were identified which were differentially expressed in OA SF. S100-A10, a calcium binding protein was upregulated in OA and validated with western blotting and immunohistochemistry. Several new OA specific peptide fragments (neopeptides) were identified.The protein equalisation method compressed the dynamic range of the synovial proteins identifying the most comprehensive SF proteome to date. A number of proteins were identified for the first time in SF which may be involved in the pathogenesis of OA. We identified a distinct set of proteins and neopeptides that may act as potential biomarkers to distinguish between normal and OA joints.

2022-03-02 | PXD001509 | Pride

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