Single cell RNA-seq analysis identifies ferroptotic chondrocyte cluster and reveals TRPV1 as an anti-ferroptotic target in osteoarthritis.

Animals Cartilage, Articular / metabolism Chondrocytes / metabolism Glutathione / metabolism Humans Iron / metabolism Mice Osteoarthritis / drug therapy Phospholipid Hydroperoxide Glutathione Peroxidase / genetics Sequence Analysis, RNA TRPV Cation Channels / genetics tert-Butylhydroperoxide / metabolism

Ferroptosis GPX4 Osteoarthritis Single cell RNA sequencing TRPV1

Journal

EBioMedicine

ISSN: 2352-3964

Titre abrégé: EBioMedicine

Pays: Netherlands

ID NLM: 101647039

Informations de publication

Date de publication:
Oct 2022

Historique:

received: 15 04 2022

revised: 25 08 2022

accepted: 25 08 2022

pubmed: 23 9 2022

medline: 19 10 2022

entrez: 22 9 2022

Statut: ppublish

Résumé

Osteoarthritis (OA) is the most common degenerative joint disease primarily characterized by cartilage destruction. The aim of this study was to investigate the role, molecular characteristics and potential therapeutic target of chondrocyte ferroptosis in the pathogenesis of OA. The expression of ferroptotic hallmarks (iron and lipid peroxidation accumulation, glutathione deletion) were analyzed in paired intact and damaged cartilages from OA patients. Single cell RNA sequencing (scRNA-seq) analysis was performed on 17,638 chondrocytes to verify the presence, investigate the molecular signatures and unveil the potential therapeutic target of ferroptotic chondrocyte cluster in human OA cartilages. Destabilization of medial meniscus (DMM)-induced OA model and tert-butyl hydroperoxide (TBHP)-treated primary mouse chondrocytes and human cartilage explants were used to evaluate the protective effect of pharmacologically activated transient receptor potential vanilloid 1 (TRPV1). The downstream molecular mechanisms of TRPV1 was further investigated in glutathione peroxidase 4 (Gpx4) heterozygous genetic deletion mice (Gpx4 The concentrations of iron and lipid peroxidation and the expression of ferroptotic drivers in the damaged areas of human OA cartilages were significantly higher than those in the intact cartilage. scRNA-seq analysis revealed a chondrocyte cluster characterized by preferentially expressed ferroptotic hallmarks and genes, namely ferroptotic chondrocyte cluster. Comprehensive gene set variation analysis revealed TRPV1 as an anti-ferroptotic target in human OA cartilage. Pharmacological activation of TRPV1 significantly abrogated cartilage degeneration by protecting chondrocytes from ferroptosis. Mechanistically, TRPV1 promoted the expression of GPX4, and its anti-ferroptotic role was largely mitigated in the OA model of Gpx4 TRPV1 activation protects chondrocytes from ferroptosis and ameliorates OA progression by upregulating GPX4. National Key R&D Program of China (2018YFC1105904), Key Program of NSFC (81730067), National Science Foundation of China (81772335, 81941009, 81802196), Natural Science Foundation of Jiangsu Province, China (BK20180127), Jiangsu Provincial Key Medical Talent Foundation, Six Talent Peaks Project of Jiangsu Province (WSW-079).

Sections du résumé

BACKGROUND BACKGROUND

METHODS METHODS

The expression of ferroptotic hallmarks (iron and lipid peroxidation accumulation, glutathione deletion) were analyzed in paired intact and damaged cartilages from OA patients. Single cell RNA sequencing (scRNA-seq) analysis was performed on 17,638 chondrocytes to verify the presence, investigate the molecular signatures and unveil the potential therapeutic target of ferroptotic chondrocyte cluster in human OA cartilages. Destabilization of medial meniscus (DMM)-induced OA model and tert-butyl hydroperoxide (TBHP)-treated primary mouse chondrocytes and human cartilage explants were used to evaluate the protective effect of pharmacologically activated transient receptor potential vanilloid 1 (TRPV1). The downstream molecular mechanisms of TRPV1 was further investigated in glutathione peroxidase 4 (Gpx4) heterozygous genetic deletion mice (Gpx4

FINDINGS RESULTS

The concentrations of iron and lipid peroxidation and the expression of ferroptotic drivers in the damaged areas of human OA cartilages were significantly higher than those in the intact cartilage. scRNA-seq analysis revealed a chondrocyte cluster characterized by preferentially expressed ferroptotic hallmarks and genes, namely ferroptotic chondrocyte cluster. Comprehensive gene set variation analysis revealed TRPV1 as an anti-ferroptotic target in human OA cartilage. Pharmacological activation of TRPV1 significantly abrogated cartilage degeneration by protecting chondrocytes from ferroptosis. Mechanistically, TRPV1 promoted the expression of GPX4, and its anti-ferroptotic role was largely mitigated in the OA model of Gpx4

INTERPRETATION CONCLUSIONS

TRPV1 activation protects chondrocytes from ferroptosis and ameliorates OA progression by upregulating GPX4.

FUNDING BACKGROUND

National Key R&D Program of China (2018YFC1105904), Key Program of NSFC (81730067), National Science Foundation of China (81772335, 81941009, 81802196), Natural Science Foundation of Jiangsu Province, China (BK20180127), Jiangsu Provincial Key Medical Talent Foundation, Six Talent Peaks Project of Jiangsu Province (WSW-079).

Identifiants

DOI: 10.1016/j.ebiom.2022.104258 PMID: 36137413 PMC: PMC9494174

pubmed: 36137413

pii: S2352-3964(22)00440-6

doi: 10.1016/j.ebiom.2022.104258

pmc: PMC9494174

pii:

doi:

Substances chimiques

TRPV Cation Channels 0

TRPV1 protein, human 0

TRPV1 protein, mouse 0

tert-Butylhydroperoxide 955VYL842B

Iron E1UOL152H7

Phospholipid Hydroperoxide Glutathione Peroxidase EC 1.11.1.12

Glutathione GAN16C9B8O

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

Pagination

104258

Informations de copyright

Déclaration de conflit d'intérêts

Declaration of interests The authors have declared that no conflict of interest exists.