Biology subjects

Welting, T. J.

Publications and source records attributed to Welting, T. J..

3 recordsLinked to original sources

bioRxiv2023

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

Extracellular vesicles contribute to osteoarthritis pathogenesis through their release into joint tissues and synovial fluid. Limited studies have profiled extracellular vesicles in osteoarthritic biofluids, such as plasma and synovial fluid. Due to the potential involvement in osteoarthritis pathogenesis, synovial fluid-derived extracellular vesicles 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 early osteoarthritis, we defined the changes in matched synovial fluid and plasma-derived extracellular vesicle small non-coding RNA and protein cargo using small RNA sequencing and mass spectrometry proteomics. We explored the data with a multi-omic approach including time series clustering, factor analysis and gene enrichment interrogation. Chondrocyte signalling induced by temporal synovial fluid-derived extracellular vesicles derived from the model were analysed using luciferase-based transcription factor activity assays. Extracellular vesicle protein cargo appears to be more important during osteoarthritis progression than small non-coding RNA cargo. Cluster analysis revealed plasma-extracellular vesicles represented a time-dependant response to osteoarthritis induction, were principally derived from protein cargo and were associated with supramolecular complexes. Clusters for synovial fluid-derived extracellular vesicles were associated with an initial osteoarthritis response and represented immune/inflammatory pathways. Factor analysis revealed that plasma-derived extracellular vesicles correlated with day post induction and were primarily composed of proteins which may modulate lipid metabolism in osteoarthritis. Synovial fluid-derived extracellular vesicles significant factors represented intermediate filament and supramolecular complexes reflecting tissue repair responses to osteoarthritis induction. There was a significant interaction between time and osteoarthritis for cAMP response element, Nuclear factor-kappa B response element, serum response element and serum response factor response element reporters with a trend for osteoarthritis synovial fluid-derived EVs at later time points to have a more pronounced effect. Local and systemic osteoarthritis-associated changes in extracellular vesicle cargo profiles in this in vivo model provided a unique opportunity to understand their role in disease propagation and progression and may represent novel biomarkers to stage osteoarthritis.

molecular biology↗

bioRxiv2020

Small RNAome of ageing chondrocytes

BackgroundAgeing is one of the leading risk factors predisposing cartilage to musculoskeletal diseases, including osteoarthritis. Cumulative evidence suggests that small non-coding RNAs play a role in cartilage-related pathological changes. However, little research has been conducted on the effect of ageing on the expression of small non-coding RNAs in cartilage. By using small RNA sequencing, we investigated changes in the expression of small non-coding RNAs between young and old equine chondrocytes. MethodsChondrocytes were extracted from five young (4{+/-}1 years) and five old (17.4{+/-}1.9 years) macroscopically normal equine metacarpophalangeal joints. Following RNA extraction cDNA libraries were prepared and subjected to small RNA sequencing using the Illumina MiSeq platform. Differential expression analysis was performed in R using package DESeq2. For tRNA fragment analysis, tRNA reads were aligned to horse tRNA sequences using Bowtie2 version 2.2.5. Selected microRNA and small nucleolar RNA findings were validated using qRT-PCR in an extended cohort of equine chondrocytes. tRNA fragments were further investigated in low and high grade OA human cartilage tissue. ResultsIn total, 83 sncRNAs were differentially expressed between young and old equine chondrocytes, including microRNAs, snoRNAs, snRNAs and tRNAs. Of these, 34 were expressed higher and 49 were expressed lower in old chondrocytes compared to young. qRT-PCR analysis confirmed findings in an extended cohort of equine chondrocytes. Ingenuity Pathway Analysis of differentially expressed microRNAs and their predicted target genes linked them to cartilage and OA-related pathways and diseases. tRNA fragment analysis revealed that tiRNA-5035-GluCTC and tiRNA-5031-GluCTC-1 were reduced in both high grade OA human cartilage and old equine chondrocytes. ConclusionFor the first time, we have measured the effect of ageing on the expression of small non-coding RNAs in equine chondrocytes. Changes were detected in a number of different sncRNA species, including microRNAs, small nucleolar RNAs and tRNA fragments. This study supports a role for small non-coding RNAs in ageing cartilage and their potential involvement in age-related cartilage diseases.

molecular biology↗

bioRxiv2020

Equine synovial fluid small non-coding RNA signatures in early osteoarthritis

BackgroundOsteoarthritis remains one of the greatest causes of morbidity and mortality in the equine population. The inability to detect pre-clinical changes in osteoarthritis has been a significant impediment to the development of effective therapies against this disease. Synovial fluid represents a potential source of disease-specific small non-coding RNAs (sncRNAs) that could aid in the understanding of the pathogenesis of osteoarthritis. We hypothesised that early stages of osteoarthritis would alter the expression of sncRNAs, facilitating the understanding of the underlying pathogenesis and potentially provide early biomarkers. MethodsSmall RNA sequencing was performed using synovial fluid from the metacarpophalangeal joints of both control and early osteoarthritic non-Thoroughbred horses. A group of differentially expressed sncRNAs was selected for further validation through qRT-PCR using an independent cohort of synovial fluid samples from control and early osteoarthritic horses. Bioinformatic analysis was performed in order to identify putative targets of the differentially expressed microRNAs and to explore potential associations with specific biological processes. ResultsResults revealed 22 differentially expressed sncRNAs including 13 microRNAs; miR-10a, miR-223, let7a, miR-99a, miR-23b, miR-378, miR-143 (and six novel microRNAs), four small nuclear RNAs; U2, U5, U11, U12, three small nucleolar RNAs; U13, snoR38, snord96, and one small cajal body-specific RNA; scarna3. Five sncRNAs were validated; miR-223 was significantly reduced in early OA and miR-23b, let-7a-2, snord96A and snord13 were significantly upregulated. Significant cellular functions deduced by the differentially expressed microRNAs included apoptosis (P < 0.0003), necrosis (P < 0.0009), autophagy (P < 0.0007) and inflammation (P < 0.00001). A conservatively filtered list of 57 messenger RNA targets was obtained; the top biological processes associated were regulation of cell population proliferation (P < 0.000001), cellular response to chemical stimulus (P < 0.000001) and cell surface receptor signalling pathway (P < 0.000001). ConclusionsSynovial fluid sncRNAs can be used as molecular biomarkers for early disease in equine osteoarthritic joints. The biological processes they regulate may play an important role in understanding early osteoarthritis pathogenesis. Characterising these dynamic molecular changes could provide novel insights on the process and mechanism of early osteoarthritis development and is critical for the development of new therapeutic approaches.

molecular biology↗