Anderson, James R

Castanheira, Catarina Igd

Clarke, Emily J

Clegg, Peter D

Hackl, Matthias

James, Victoria

Peffers, Mandy J

University of Liverpool Repository

EXTRACELLULAR VESICLE MICRORNA CROSSTALK BETWEEN CHONDROCYTES AND SYNOVIOCYTES Extracellular vesicles (EVs) are crucial in cell-to-cell communication and can influence joint homeostasis and contribute to osteoarthritis. The aim of this study was to describe a novel technique for the investigation of the EV-derived small non-coding RNA (sncRNA) crosstalk between equine chondrocytes and synoviocytes in vitro, using RNA tracking and sequencing.Healthy chondrocytes (n=8) and synoviocytes (n=9) were incubated with a 5-Ethynyl Uridine tag, labelling newly synthesised RNA. EVs were isolated from culture media using size exclusion chromatography and characterised by nanoparticle tracking. Recipient cells (chondrocytes, n=5; synoviocytes, n=4) were incubated with tagged EVs; chondrocytes were incubated with synoviocyte-derived EVs, and recipient synoviocytes were incubated with chondrocyte-derived EVs.  RNA was extracted and 5-EU labelled RNA recovered from recipient cells using an affinity recovery kit, then sequenced. Differential expression was undertaken using edgeR v3.32. Results were used for pathway analysis and target prediction.198 miRNAs and 269 snoRNAs were identified in synoviocytes, corresponding to the 5-EU labelled RNA from chondrocyte EVs. 213 miRNAs and 265 snoRNAs were identified in chondrocytes, corresponding to the 5-EU labelled RNA from synoviocyte EVs. Differential expression analysis revealed nine differentially expressed miRNAs and seven snoRNAs. Pathway analysis revealed significant links to migration of cells, apoptosis, cell viability of connective tissue cell and inflammation of joint.Sequencing data revealed distinct miRNA patterns between chondrocyte and synoviocyte-derived EVs. Pathway analysis revealed that the differentially expressed miRNAs can be related to cell viability and inflammation, and some of these miRNAs have also been suggested as potential biomarkers for osteoarthritis.

Extracellular vesicle microRNA crosstalk between chondrocytes and sysnoviocytes- an in vitro approach

http://livrepository.liverpool.ac.uk/3177861/1/EXTRACELLULAR%20VESICLE%20MICRORNA%20CROSSTALK%20BETWEEN%20CHONDROCYTES%20AND%20SYNOVIOCYTES.docx

Extracellular vesicle microRNA crosstalk between chondrocytes and sysnoviocytes- an in vitro approach

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