Unique diagnostic signatures of concussion in the saliva of male athletes: the Study of Concussion in Rugby Union through MicroRNAs (SCRUM)
Valentina Di Pietro, Patrick O'Halloran, Callum N Watson, Ghazala Begum, Animesh Acharjee, Kamal M Yakoub, Conor Bentley, David J Davies, Paolo Iliceto, Gabriella Candilera, David K Menon, Matthew J Cross, Keith A Stokes, Simon Pt Kemp, Antonio Belli, Valentina Di Pietro, Patrick O'Halloran, Callum N Watson, Ghazala Begum, Animesh Acharjee, Kamal M Yakoub, Conor Bentley, David J Davies, Paolo Iliceto, Gabriella Candilera, David K Menon, Matthew J Cross, Keith A Stokes, Simon Pt Kemp, Antonio Belli
Abstract
Objective: To investigate the role of salivary small non-coding RNAs (sncRNAs) in the diagnosis of sport-related concussion.
Methods: Saliva was obtained from male professional players in the top two tiers of England's elite rugby union competition across two seasons (2017-2019). Samples were collected preseason from 1028 players, and during standardised head injury assessments (HIAs) at three time points (in-game, post-game, and 36-48 hours post-game) from 156 of these. Samples were also collected from controls (102 uninjured players and 66 players sustaining a musculoskeletal injury). Diagnostic sncRNAs were identified with next generation sequencing and validated using quantitative PCR in 702 samples. A predictive logistic regression model was built on 2017-2018 data (training dataset) and prospectively validated the following season (test dataset).
Results: The HIA process confirmed concussion in 106 players (HIA+) and excluded this in 50 (HIA-). 32 sncRNAs were significantly differentially expressed across these two groups, with let-7f-5p showing the highest area under the curve (AUC) at 36-48 hours. Additionally, a combined panel of 14 sncRNAs (let-7a-5p, miR-143-3p, miR-103a-3p, miR-34b-3p, RNU6-7, RNU6-45, Snora57, snoU13.120, tRNA18Arg-CCT, U6-168, U6-428, U6-1249, Uco22cjg1,YRNA_255) could differentiate concussed subjects from all other groups, including players who were HIA- and controls, immediately after the game (AUC 0.91, 95% CI 0.81 to 1) and 36-48 hours later (AUC 0.94, 95% CI 0.86 to 1). When prospectively tested, the panel confirmed high predictive accuracy (AUC 0.96, 95% CI 0.92 to 1 post-game and AUC 0.93, 95% CI 0.86 to 1 at 36-48 hours).
Conclusions: SCRUM, a large prospective observational study of non-invasive concussion biomarkers, has identified unique signatures of concussion in saliva of male athletes diagnosed with concussion.
Keywords: brain; concussion; contact sports; diagnosis; trauma.
Conflict of interest statement
Competing interests: AB and VDP are founding members and shareholders of Marker Diagnostics, a spinout company of the University of Birmingham. GB and POH are currently employed by Marker Diagnostics. SK and KS are employed by the RFU, the National Governing Body for the game in England. The RFU has a financial interest in the intellectual property connected to the biomarkers here described. MC is employed by Premiership Rugby.
© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY. Published by BMJ.
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