WWP2 confers risk to osteoarthritis by affecting cartilage matrix deposition via hypoxia associated genes

M Tuerlings; G M C Janssen; I Boone; M van Hoolwerff; A Rodriguez Ruiz; E Houtman; H E D Suchiman; R J P van der Wal; R G H H Nelissen; R Coutinho de Almeida; P A van Veelen; Y F M Ramos; I Meulenbelt

doi:10.1016/j.joca.2022.09.009

WWP2 confers risk to osteoarthritis by affecting cartilage matrix deposition via hypoxia associated genes

Osteoarthritis Cartilage. 2023 Jan;31(1):39-48. doi: 10.1016/j.joca.2022.09.009. Epub 2022 Oct 5.

Authors

Affiliations

¹ Dept. of Biomedical Data Sciences, Section Molecular Epidemiology, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
² Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
³ Dept. of Biomedical Data Sciences, Section Molecular Epidemiology, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
⁴ Dept. of Biomedical Data Sciences, Section Molecular Epidemiology, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
⁵ Dept. of Biomedical Data Sciences, Section Molecular Epidemiology, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
⁶ Dept. of Biomedical Data Sciences, Section Molecular Epidemiology, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
⁷ Dept. of Biomedical Data Sciences, Section Molecular Epidemiology, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
⁸ Dept. Orthopaedics, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
⁹ Dept. Orthopaedics, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
¹⁰ Dept. of Biomedical Data Sciences, Section Molecular Epidemiology, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
¹¹ Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
¹² Dept. of Biomedical Data Sciences, Section Molecular Epidemiology, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].
¹³ Dept. of Biomedical Data Sciences, Section Molecular Epidemiology, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: [email protected].

PMID: 36208715
DOI: 10.1016/j.joca.2022.09.009

Abstract

Objective: To explore the co-expression network of the osteoarthritis (OA) risk gene WWP2 in articular cartilage and study cartilage characteristics when mimicking the effect of OA risk allele rs1052429-A on WWP2 expression in a human 3D in vitro model of cartilage.

Method: Co-expression behavior of WWP2 with genes expressed in lesioned OA articular cartilage (N = 35 samples) was explored. By applying lentiviral particle mediated WWP2 upregulation in 3D in vitro pellet cultures of human primary chondrocytes (N = 8 donors) the effects of upregulation on cartilage matrix deposition was evaluated. Finally, we transfected primary chondrocytes with miR-140 mimics to evaluate whether miR-140 and WWP2 are involved in similar pathways.

Results: Upon performing Spearman correlations in lesioned OA cartilage, 98 highly correlating genes (|ρ| > 0.7) were identified. Among these genes, we identified GJA1, GDF10, STC2, WDR1, and WNK4. Subsequent upregulation of WWP2 on 3D chondrocyte pellet cultures resulted in a decreased expression of COL2A1 and ACAN and an increase in EPAS1 expression. Additionally, we observed a decreased expression of GDF10, STC2, and GJA1. Proteomics analysis identified 42 proteins being differentially expressed with WWP2 upregulation, which were enriched for ubiquitin conjugating enzyme activity. Finally, upregulation of miR-140 in 2D chondrocytes resulted in significant upregulation of WWP2 and WDR1.

Conclusions: Mimicking the effect of OA risk allele rs1052429-A on WWP2 expression initiates detrimental processes in the cartilage shown by a response in hypoxia associated genes EPAS1, GDF10, and GJA1 and a decrease in anabolic markers, COL2A1 and ACAN.

Keywords: Articular cartilage; Osteoarthritis; WWP2; miRNA-140.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cartilage, Articular* / metabolism
Cells, Cultured
Chondrocytes / metabolism
Humans
Hypoxia
MicroRNAs* / metabolism
Osteoarthritis* / genetics
Osteoarthritis* / metabolism
Ubiquitin-Protein Ligases / metabolism

Substances

MicroRNAs
WWP2 protein, human
Ubiquitin-Protein Ligases
Mirn140 microRNA, human