Dataset Information

Global Extracellular Vesicle Proteome Changes Across Time Upon Intraarticular Treatment of Mesenchymal Stromal Cells in an In Vivo model of Equine Osteoarthritis

ABSTRACT: Equine Osteoarthritis (OA) is a heterogeneous, degenerative disease of the musculoskeletal system with multifactorial causation, characterised by joint metabolic imbalance. Mesenchymal stromal cell therapy (MSC) is a form of regenerative medicine that utilises MSC properties to repair damaged tissues. MSCs have been described as acting via the paracrine signalling of secreted factors. Despite its wide use in veterinary clinical practices, the exact mechanism of action of MSCs has not been fully characterised. Here, we have characterised synovial fluid extracellular vesicles (EVs) from control, osteoarthritic and MSC treated animals in order to gain insights into this mechanism. An in vivo, carpal osteochondral fragment model of equine OA was used for this study. Six horses underwent surgical intervention. All horses enrolled in the study were female trotter horses between the ages of 4 and 7. The contralateral limb of each horse served as a sham control. 69 synovial fluid samples were collected via aseptic arthrocentesis at day 0, 18, 21, 28, 35, and 70. Allogenic mesenchymal stromal cell therapy from male donors was injected into the OA afflicted joint after day 18. Synovial fluid (200µl) was Hyalaronidase treated (1µg/ml) and EVs were isolated using differential ultracentrifugation. EVs were characterised in collaboration with Nanoview Biosciences, whereby the Exoview human tetraspanin assay was used. EV concentration, surface marker identification, fluorescent microscopy and tetraspanin colocalization analysis was performed using pooled samples reflecting experimental groups - control, OA and OA including MSCs - across time.

INSTRUMENT(S): TripleTOF 6600

ORGANISM(S): Equus Caballus (horse)

TISSUE(S): Extracellular Vesicle, Synovial Tissue

DISEASE(S): Osteoarthritis

SUBMITTER: Rosalind Jenkins

LAB HEAD: Roz Jenkins

PROVIDER: PXD035303 | Pride | 2023-03-11

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	D__Sciex_data_TT6600_data_MS_Data_TT6600_Feb_2022_EJC_SF_EVs_1_101_SWATH_140222_wiff_dia.quant	Other
	D__Sciex_data_TT6600_data_MS_Data_TT6600_Feb_2022_EJC_SF_EVs_1_102_SWATH_140222_wiff_dia.quant	Other
	D__Sciex_data_TT6600_data_MS_Data_TT6600_Feb_2022_EJC_SF_EVs_1_103_SWATH_140222_wiff_dia.quant	Other
	D__Sciex_data_TT6600_data_MS_Data_TT6600_Feb_2022_EJC_SF_EVs_1_108_SWATH_140222_wiff_dia.quant	Other
	D__Sciex_data_TT6600_data_MS_Data_TT6600_Feb_2022_EJC_SF_EVs_1_112_SWATH_110222_wiff_dia.quant	Other

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Publications

Temporal extracellular vesicle protein changes following intraarticular treatment with integrin α10β1-selected mesenchymal stem cells in equine osteoarthritis.

Clarke Emily J EJ Johnson Emily E Caamaño Gutierrez Eva E Andersen Camilla C Berg Lise C LC Jenkins Rosalind E RE Lindegaard Casper C Uvebrant Kristina K Lundgren-Åkerlund Evy E Turlo Agnieszka A James Victoria V Jacobsen Stine S Peffers Mandy J MJ

Frontiers in veterinary science 20221124

<h4>Introduction</h4>Equine osteoarthritis (OA) is a heterogeneous, degenerative disease of the musculoskeletal system with multifactorial causation, characterized by a joint metabolic imbalance. Extracellular vesicles are nanoparticles involved in intracellular communication. Mesenchymal stem cell (MSC) therapy is a form of regenerative medicine that utilizes their properties to repair damaged tissues. Despite its wide use in veterinary practice, the exact mechanism of action of MSCs is not ful ...[more]

PMID: 36504839

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Project description:Objective: To quantify gene expression changes in the synovium of osteoarthritis-affected joints in an equine metacarpophalangeal joint (MCPJ) chip model specifically designed to recapitulate early post-traumatic osteoarthritis (PTOA). Design: Synovial samples were collected arthroscopically from the MCPJ of 11 adult horses before (pre-OA) and after (OA) surgical induction of osteoarthritis and from sham-operated joints. Briefly, in this model, an osteochondral fragment is created in one randomly chosen MCPJ at the proximal dorsomedial aspect of the first phalanx. The fragment is replaced in the fragment bed after creation so that subchondral bone is not exposed to the opposing cartilage surface. The opposite MCPJ is sham-operated. After a two week recover period, the horses are treadmill-exercised for 14 weeks and then the osteochondral fragment is removed (16 weeks after creation). Synovial tissue samples were collected arthroscopically from the MCPJ of eleven adult horses before (pre-OA) and 16 weeks after (OA) experimental induction of OA as well as from the sham-operated joints (sham). After sequencing of synovial RNA, Salmon was used to quasi-map reads to the reference genome and quantify transcript abundances. Differential expression analysis was performed with the limma-treat method using a fold-change cutoff of 1.1. Functional annotation was performed with PANTHER and Reactome at FDR < 0.05. Results: RNA was successfully extracted from 28 samples (6 pre-OA, 11 OA, 11 sham). Sequencing yielded 15.7-29.4 million paired-end reads per sample. “Sham” and “pre-OA” were not different and were grouped. 321 genes were upregulated and 351 genes were downregulated in OA synovium compared to unaffected. Gene ontology (GO) terms related to extracellular matrix (ECM) organization and growth factor binding were overrepresented among differentially expressed genes. There were 20 significantly enriched pathways; these included pathways involved in ECM turnover, O-glycosylation of TSR domain-containing proteins, and growth factor signaling. Conclusions: Most enriched pathways and overrepresented GO terms reflect a state of high metabolic activity and tissue turnover in OA-affected tissue, suggesting efforts at healing and restoring homeostasis. Limitations of this study include a small sample size and capture of a single point post-injury. Differentially expressed genes falling within key pathways may represent potential diagnostic markers or therapeutic targets for PTOA.