Mandy Perffers
Research Lead, Department of Musculoskeletal Science and Ageing, Institute of Life Course and Medical Sciences, University of Liverpool, United Kingdon
Mandy Perffers obtained a degree in Animal Science at the University of Leeds, and then undertook her veterinary degree at The Royal Veterinary College, University of London and qualified as a veterinarian in 1995. Following an internship in reproduction at Glasgow University she then spent 11 years in industry and private practice before undertaking a PhD supported by the Wellcome Trust on ‘Proteomic and transcriptomic signatures of cartilage ageing and disease’. Her fellowship continued by studying ‘A Systems Biology Approach to Musculoskeletal Ageing’. She then obtained a Wellcome Trust Clinical Intermediate Fellow studying the role of small nucleolar RNAs in cartilage ageing and disease as well as the potential use of extracellular vesicles to treat equine osteoarthritis. Her research group at Liverpool is a mix of vets, scientists at master, PhD and postdoctoral level studying the role of epigenetics in musculoskeletal diseases in man, dogs and horses. They are additionally interested in the role of extracellular vesicles in osteoarthritis as biomarkers and potential treatments. Much of her research has started with ‘omic’ discovery experiments to identify molecules for mechanistic studies or biomarkers. They utilise many forms of ‘omics’ technologies in our research including mass spectrometry proteomics (label-free quantification, absolute quantification and mass spectrometry imaging), nuclear magnetic resonance metabolomics, microarray, RNASeq, small RNASeq, QuantSeqReverse, ribosomal profiling and DNA methylation arrays. She is currently section editor for omics for the Equine Veterinary Journal.
Small non-coding RNA landscape of extracellular vesicles from an
experimental model of equine osteoarthritis
Joint tissues release extracellular vesicles (EVs) that potentially sustain joint homeostasis and contribute to osteoarthritis (OA) pathogenesis. EVs are putative novel therapeutics for OA, and transport biologically active molecules (including small non-coding RNAs (SNCRNAs)) between cells. This study identified altering SNCRNA cargo in EVs in OA which may act as early diagnostic markers and treatment targets.
OA was surgically induced in four skeletally mature Standardbred horses using an osteochondral fragment model in the left middle carpal joint. The right joint underwent sham surgery. Synovial fluid (SF)and plasma were obtained weekly throughout the 70-day study. EVs were isolated using size exclusion chromatography and characterised using nanoparticle tracking (Nanosight), and exosome fluorescence detection and tetraspanin phenotyping (Exoview). RNA was extracted from EVs derived from SF (sham and OA joints) and plasma collected at days 10, 35, 42, 49, 56, 63, and subjected to small RNA sequencing on a NovaSeq SP100 flow cell (Illumina).
Nanosight-derived EV characteristics of size and concentration were not significantly different following disease induction. The diameter of the temporal population of plasma and SF-derived exosomes changed significantly for both CD9 and CD81 following OA induction with significant temporal, and disease-related changes in CD63 and CD81 tetraspanin protein expressin in plasma and SF.
The differential expression temporally of seven microRNAs in plasma and synovial fluid-derived extracellular vesicles; eca-miR-451, eca-miR-25, eca-miR-215, eca-miR-92a, eca-miR-let-7c, eca-miR-486-5p, eca-miR-23a and four snoRNAs; U3, snord15, snord46, snord58 represent potential biomarkers for early osteoarthritis. Bioinformatics analysis of the differentially expressed microRNAs in synovial fluid highlighted that in early osteoarthritis these related to the inhibition of cell cycle, cell cycle progression, DNA damage and cell proliferation, but increased cell viability and differentiation of stem cells.
Plasma and synovial fluid-derived extracellular vesicle small non-coding signatures have been established for the first time in a temporal model of osteoarthritis. These could serve as novel biomarkers for the evaluation of osteoarthritis progression or act as potential therapeutic targets.
OA was surgically induced in four skeletally mature Standardbred horses using an osteochondral fragment model in the left middle carpal joint. The right joint underwent sham surgery. Synovial fluid (SF)and plasma were obtained weekly throughout the 70-day study. EVs were isolated using size exclusion chromatography and characterised using nanoparticle tracking (Nanosight), and exosome fluorescence detection and tetraspanin phenotyping (Exoview). RNA was extracted from EVs derived from SF (sham and OA joints) and plasma collected at days 10, 35, 42, 49, 56, 63, and subjected to small RNA sequencing on a NovaSeq SP100 flow cell (Illumina).
Nanosight-derived EV characteristics of size and concentration were not significantly different following disease induction. The diameter of the temporal population of plasma and SF-derived exosomes changed significantly for both CD9 and CD81 following OA induction with significant temporal, and disease-related changes in CD63 and CD81 tetraspanin protein expressin in plasma and SF.
The differential expression temporally of seven microRNAs in plasma and synovial fluid-derived extracellular vesicles; eca-miR-451, eca-miR-25, eca-miR-215, eca-miR-92a, eca-miR-let-7c, eca-miR-486-5p, eca-miR-23a and four snoRNAs; U3, snord15, snord46, snord58 represent potential biomarkers for early osteoarthritis. Bioinformatics analysis of the differentially expressed microRNAs in synovial fluid highlighted that in early osteoarthritis these related to the inhibition of cell cycle, cell cycle progression, DNA damage and cell proliferation, but increased cell viability and differentiation of stem cells.
Plasma and synovial fluid-derived extracellular vesicle small non-coding signatures have been established for the first time in a temporal model of osteoarthritis. These could serve as novel biomarkers for the evaluation of osteoarthritis progression or act as potential therapeutic targets.
