RT Journal Article
SR Electronic
T1 Small Non-coding RNAome of ageing chondrocytes
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.17.156927
DO 10.1101/2020.06.17.156927
A1 Panagiotis Balaskas
A1 Jonathan A. Green
A1 Tariq M. Haqqi
A1 Philip Dyer
A1 Yalda A. Kharaz
A1 Yongxiang Fang
A1 Xuan Liu
A1 Tim J.M. Welting
A1 Mandy J. Peffers
YR 2020
UL http://biorxiv.org/content/early/2020/06/20/2020.06.17.156927.abstract
AB Background Ageing 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.Methods Chondrocytes 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.Results In 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.Conclusion For 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.Competing Interest StatementT.J.M Welting is listed as inventors on patents WO2017178251 and WO2017178253. T.J.M Welting has shares in Chondropeptix. The remaining authors declare that they have no competing interests.