Ribosomal RNA-based epitranscriptomic regulation of chondrocyte translation and proteome in osteoarthritis


Chabronova, A ORCID: 0000-0002-9733-444X, van den Akker, GGH, Housmans, BAC, Caron, MMJ, Cremers, A, Surtel, DAM, Wichapong, K, Peffers, MMJ, van Rhijn, LW, Marchand, V et al (show 2 more authors) (2023) Ribosomal RNA-based epitranscriptomic regulation of chondrocyte translation and proteome in osteoarthritis. OSTEOARTHRITIS AND CARTILAGE, 31 (3). pp. 374-385.

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Abstract

<h4>Objective</h4>Osteoarthritis-related cartilage extracellular matrix remodeling is dependent on changes in chondrocyte protein expression. Yet, the role of ribosomes in chondrocyte translation regulation is unknown. In this exploratory study, we investigated ribosomal RNA (rRNA) epitranscriptomic-based ribosome heterogeneity in human articular chondrocytes and its relevance for osteoarthritis.<h4>Methods</h4>Sequencing-based rRNA 2'-O-methylation profiling analysis (RiboMethSeq) was performed on non-OA primary human articular chondrocytes (n = 5) exposed for 14 days to osteoarthritic synovial fluid (14 donors, pooled, 20% v/v). The SW1353 SNORD71 KO cell pool was generated using LentiCRISPRv2/Cas9. The mode of translation initiation and fidelity were determined by dual-luciferase reporters. The cellular proteome was analyzed by LC-MS/MS and collagen type I protein expression was evaluated by immunoblotting. Loading of COL1A1 mRNA into polysomes was determined by sucrose gradient ultracentrifugation and fractionation.<h4>Results</h4>We discovered that osteoarthritic synovial fluid instigates site-specific changes in the rRNA 2'-O-me profile of primary human articular chondrocytes. We identified five sites with differential 2'-O-me levels. The 2'-O-me status of 5.8S-U14 (one of identified differential 2'-O-me sites; decreased by 7.7%, 95% CI [0.9-14.5%]) was targeted by depleting the level of its guide snoRNA SNORD71 (50% decrease, 95% CI [33-64%]). This resulted in an altered ribosome translation modus (e.g., CrPV IRES, FC 3, 95% CI [2.2-4.1]) and promoted translation of COL1A1 mRNA which led to increased levels of COL1A1 protein (FC 1.7, 95% CI [1.3-2.0]).<h4>Conclusions</h4>Our data identify a novel concept suggesting that articular chondrocytes employ rRNA epitranscriptomic mechanisms in osteoarthritis development.

Item Type: Article
Uncontrolled Keywords: Chondrocytes, Collagen type I, 5, 8S rRNA, Ribosome, Preferential translation, rRNA epitranscriptomic regulation
Divisions: Faculty of Health and Life Sciences
Faculty of Health and Life Sciences > Institute of Life Courses and Medical Sciences
Depositing User: Symplectic Admin
Date Deposited: 10 Jul 2023 14:05
Last Modified: 10 Jul 2023 14:05
DOI: 10.1016/j.joca.2022.12.010
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3171600
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