Multi-Omic Temporal Landscape of Plasma and Synovial Fluid-Derived Extracellular Vesicles Using an Experimental Model of Equine Osteoarthritis

Abstract

Extracellular vesicles (EVs) contribute to osteoarthritis pathogenesis through their release into joint tissues and synovial fluid. Synovial fluid-derived EVs have the potential to be direct biomarkers in the causal pathway of disease but also enable understanding of their role in disease progression. Utilizing a temporal model of osteoarthritis, we defined the changes in matched synovial fluid and plasma-derived EV small non-coding RNA and protein cargo using sequencing and mass spectrometry. Data exploration included time series clustering, factor analysis and gene enrichment interrogation. Chondrocyte signalling was analysed using luciferase-based transcription factor activity assays. EV protein cargo appears to be more important during osteoarthritis progression than small non-coding RNAs. Cluster analysis revealed plasma-EVs represented a time-dependent response to osteoarthritis induction associated with supramolecular complexes. Clusters for synovial fluid-derived EVs were associated with initial osteoarthritis response and represented immune/inflammatory pathways. Factor analysis for plasma-derived EVs correlated with day post-induction and were primarily composed of proteins modulating lipid metabolism. Synovial fluid-derived EVs factors represented intermediate filament and supramolecular complexes reflecting tissue repair. There was a significant interaction between time and osteoarthritis for CRE, NFkB, SRE, SRF with a trend for osteoarthritis synovial fluid-derived EVs at later time points to have a more pronounced effect.

Plasma and synovial fluid extracellular vesicles display altered microRNA profiles in horses with naturally occurring post-traumatic osteoarthritis: an exploratory study

“…Bioinformatics analysis was performed to analyze the biological processes and pathways affected by the DE miRNAs and their putative mRNA targets in order to enhance our understanding of the roles of the dysregulated miR-NAs in the pathogenesis of PTOA. Several plasma EV DE miRNAs identified in our study, including miR-let7, miR-29, miR-144, and miR-199, and miR-1307, were similarly altered in studies 12,13 evaluating the temporal transcriptomic profile of plasma EVs in an equine model of induced OA. Furthermore, miR-95, miR-149, miR-181, miR-193, miR-195, miR-199, and miR-411 identified in our study were recognized as miRNAs and exosomal miRNAs modified in humans with OA.…”

“…Our study identified synovial fluid EV DE miR-NAs, including miR-10, miR-29, miR-34, miR-99, miR-107, miR-126, miR-144, miR-146, miR-215, and miR-486, that were also altered in studies 12,13,27 evaluating the temporal transcriptomic profile of synovial fluid EVs in an equine models of naturally occurring and induced OA. In addition, miR-21, miR-29, miR-34, miR-101, miR-105, miR-127, miR-130, miR-144, miR-146, miR-195, miR-199, miR-411, and miR-486 from our study were recognized as miRNAs and exosomal miRNAs modified in humans with OA.…”

“…Significant differences have been characterized in the EV miRNA profile of plasma and synovial fluid obtained from humans and horses with and without osteoarthritis (OA). [11][12][13] In vitro studies have found that inflamed synoviocytes and chondrocytes produce EVs capable of rapidly and markedly upregulating proinflammatory cytokine cascades, including matrix metalloproteinases (MMPs) and aggrecanases, that degrade the extracellular matrix and decrease the biomechanical integrity of articular cartilage. 10 Several studies have characterized EV fatty acid signatures, 14 hyaluronic acid content, 15 and infrared spectra 16 in horses with naturally occurring OA; however, to the authors' knowledge, EV transcriptomes have only been characterized in equine models of induced OA.…”

“…10 Several studies have characterized EV fatty acid signatures, 14 hyaluronic acid content, 15 and infrared spectra 16 in horses with naturally occurring OA; however, to the authors' knowledge, EV transcriptomes have only been characterized in equine models of induced OA. 12,13 The present exploratory study aimed to characterize and perform transcriptomic profiling of plasma-and synovial fluid−derived EVs collected from horses with and without naturally occurring PTOA. We hypothesized that plasma and synovial fluid from horses with PTOA would contain EVs with significantly different subtypes and transcriptomes than horses without PTOA.…”

Implementation of multiomic mass spectrometry approaches for the evaluation of human health following environmental exposure

“…In several brain diseases, including multiple sclerosis and spinal muscular atrophy, effective diseasemodifying treatments can normalize NfL levels, and reduction in NfL levels is associated with the clinical effectiveness of the treatment. 150,151 Regarding inflammatory diseases, proteomics of synovial fluid and plasma revealed that there is a mild time series pattern of expression during osteoarthritis progression as reported by Anderson et al 152 At the initial induction of osteoarthritis, there was a decrease in proteins responsible for signal transduction and regulation of signaling events at day 10, followed by an increase at day 63. On day 10, there was also an initial response in proteins that are responsible for conducting immune system responses.…”