Skip to main content
NCBI home page
Cellular and Molecular Life Sciences: CMLS logoLink to Cellular and Molecular Life Sciences: CMLS
. 2026 May 13;83(1):273. doi: 10.1007/s00018-026-06158-5

Deficiency of the m6A reader IGF2BP2 mediates cellular senescence of chondrocytes and triggers cartilage degeneration

Zhi-Han Shen 1,2,3,#, Xu-Ran Li 4,#, Jian Lu 5,#, Po-Lin Liu 1,2,3, Shu-Hang He 1,2,3, Qing-Song Deng 1,2,3, Ang Wang 1,2,3, Ren Zhang 6, Shi-Cong Tao 1,2,✉, Shang-Chun Guo 1,2,3,✉
PMCID: PMC13342008  PMID: 42126425

Abstract

Accumulating evidence has identified N6-methyladenosine (m6A) regulation in the progression of age-related diseases. However, whether the m6A reader protein IGF2BP2 affects the occurrence of cartilage degeneration is still unknown. Western blotting revealed dose-dependent IGF2BP2 attenuation with increasing IL-1β concentrations in C28/I2 chondrocytes. Downregulation of IGF2BP2 caused impaired glycolysis and abnormal mitochondrial function, which ultimately resulted in senescence and disturbance of extracellular matrix homeostasis. Overexpressing HIF-1α at the cellular level in chondrocytes partially rescued mitochondrial dysfunction and senescence triggered by IGF2BP2 deficiency and thus alleviated the progression of cartilage degeneration. In vivo, chondrocyte-specific Igf2bp2-knockout mice (Col2a1-CreERT; Igf2bp2flox/flox) developed cartilage degeneration. Our study, for the first time, reveals the essential role of the m6A reader IGF2BP2 in the pathogenesis of cartilage degeneration through downregulation of HIF-1α and extends the potential role of the PINK1/Parkin pathway, downregulation of which leads to the onset of mitochondrial dysfunction and senescence in the progression of articular cartilage loss.

Supplementary Information

The online version contains supplementary material available at 10.1007/s00018-026-06158-5.

Keywords: m6A reader, cellular senescence, chondrocytes, cartilage degeneration

Introduction

Cartilage degenerative disorders, including osteoarthritis (OA), have become a global health concern, affecting over 250 million people worldwide [1]. Current, treatment options for cartilage degenerative disorders include pharmacological therapy and surgery such as microfracture, autologous chondrocyte implantation, and joint replacement [2], none of which possess the ability to reverse the extracellular matrix (ECM) imbalance [3]. A good understanding of the mechanisms underlying ECM degeneration in cartilage is essential to identify new therapeutic targets and develop effective drugs that slow the progression of cartilage degeneration.

Articular cartilage, functions to withstand mechanical stress and minimize friction, thus ensuring smooth joint movement. It is composed of a significant amount of ECM and a limited number of chondrocytes [4], the only resident cell type in mature cartilage, which play a pivotal role in regulating catabolic and anabolic factors, maintaining a delicate balance between ECM synthesis and degradation [5]. Multiple factors can disrupt this balance, leading to an imbalance between ECM anabolism and catabolism. These factors include aging, trauma, wear, and obesity, which ultimately contribute to the progressive degeneration of a joint due to the limited self-repair capacity of cartilage [6].

Senescence, one of the hallmarks of aging, is a cell state characterized by a prolonged and irreversible cell-cycle withdrawal with secretion of senescence-associated secretory phenotype (SASP) and deregulated metabolism [7, 8]. Aging is a multifaceted degenerative process that has been implicated in the pathogenesis of numerous chronic diseases, including OA [9]. It is highly probable that the thread connecting aging, senescence and OA pathology is the accumulation of senescent chondrocytes coupled with gradual changes in cellular metabolism, morphology, and function, all of which contribute to loss of cartilage homeostasis [10, 11].

N6-methyladenosine (m6A) modification is one of the most common epigenetic co-transcriptional modifications of eukaryotic RNA and is a critical regulator of mRNA stability, translocation, and translation [12]. It has recently been found to play a role in a range of both physiological and pathological conditions, including in various cancers [13] as well as in age-related diseases [14, 15].

m6A modification is reversible and is controlled by proteins known as “writers”, “erasers” and “readers” [16, 17]. The m6A readers are binding proteins which recognize and interact with the m6A modified regions to regulate RNA stability, splicing and translation [18]. There has been a substantial amount of other work published on the roles of m6A modification in disease initiation and progression [19]. However, to date there have been few studies on the role of m6A modification in OA, and none of these have focused on the m6A reader IGF2BP2.

IGF2BP2 has recently been shown to be involved in inflammation and senescence [20, 21]. Meanwhile other work has revealed interactions between the m6A reader YTHDF2 and m6A modification in influencing cartilage hypertrophy [22, 23]. Additionally, m6A modification induces macrophage polarization toward the pro-inflammatory phenotype in rheumatoid arthritis, which results in intense inflammation and cartilage degradation [24]. Nevertheless, the specific potential mechanism of IGF2BP2 in cartilage degeneration remains unclear.

In this study, the authors investigated the role of IGF2BP2 in chondrocyte senescence and cartilage degeneration, an area which has received little attention to date. We found that IGF2BP2 was significantly downregulated in OA chondrocytes. Transcriptomic, proteomic and metabolomic analyses were used to investigate the potential mechanisms of IGF2BP2 in regulating cartilage homeostasis. In vitro, IGF2BP2 knockdown in chondrocytes exacerbated senescence and decreased ECM deposition, whereas supplementation with HIF-1α had a therapeutic effect, indicating the essential role of IGF2BP2-HIF-1α in maintaining cartilage homeostasis. Moreover, IGF2BP2 deficiency impaired chondrocyte mitochondrial function through downregulation of the PINK1/Parkin pathway. Using Col2a1-CreERT; Igf2bp2flox/flox mice as an animal model, we found that IGF2BP2 deficiency in chondrocytes is a sufficient condition for the occurrence of cartilage degeneration.

Results

IGF2BP2 deficiency disrupts articular homeostasis

Emerging research implicates IGF2BP2 in modulating synovial inflammation in rheumatoid arthritis [25], with age-related Igf2bp2 functional decline observed in murine models [26]. Thus, human C28/I2 chondrocytes were stimulated with exogenous IL-1β to model OA pathology, allowing detailed analysis of IGF2BP2 expression changes. Western Blotting (WB) analysis revealed dose-dependent IGF2BP2 attenuation with increasing IL-1β concentrations (Fig. 1A).

Fig. 1.

Fig. 1

Open in a new tab

IGF2BP2 deficiency disrupts articular homeostasis. (A) Western Blotting analysis of IGF2BP2 in C28/I2 chondrocytes after 24h treatment of IL-1β. (B) qRT-PCR of IGF2BP2 levels in the C28/I2 control group, shRNA negative control (shNC) group and shIGF2BP2 group (n = 3, mean ± 95%CI). (C-D) Representative images of Toluidine blue and Safranin O staining of the control group, shNC group and shIGF2BP2 group. (E) Immunofluorescence of Collagen type II (COL II) (green), SOX9 (green), Aggrecan (green) and 4',6-diamidino-2-phenylindole (DAPI, blue) in control, shNC and shIGF2BP2 groups. Quantification of COL II-positive, SOX9-positive and Aggrecan-positive areas. Data were analyzed using one-way ANOVA and are expressed as mean with 95% confidence interval (95% CI). ANOVA: analysis of variance

Subsequently, we performed knockdown experiments using an shRNA lentiviral vector to assess IGF2BP2’s role in cartilage degeneration, shRNA lentiviral knockdown was performed in C28/I2 chondrocytes: Control (untreated), shNC (non-targeting shRNA), and shIGF2BP2 groups. Lentiviral knockdown efficiency was confirmed via qRT-PCR (Fig. 1B). Toluidine blue (TB) and Safranin O staining demonstrated decreased levels of ECM deposition in the shIGF2BP2 group compared to Control and shNC groups (Fig. 1C, D), indicating IGF2BP2’s regulatory role in ECM maintenance. Next, selected ECM synthesis genes were examined by immunofluorescent staining. As shown in Fig. 1E, the expression of ECM synthesis genes (Collagen Type II (COL Ⅱ), SRY-Box Transcription Factor 9 (SOX9) and Aggrecan (ACAN)) was lower in the shIGF2BP2 group than in the Control or shNC groups. WB analysis also confirmed decreased protein levels of COL Ⅱ and Aggrecan following IGF2BP2 inhibition (Fig. S1).

Briefly, these data imply that expression of IGF2BP2 is downregulated in osteoarthritis chondrocytes, and IGF2BP2 deficiency may disrupt cartilage homeostasis by affecting ECM anabolism.

IGF2BP2 knockdown causes disturbance of ECM homeostasis and cell cycle transition

Employing total RNA-sequencing, we further investigated the underlying mechanisms involved in disturbance of cartilage homeostasis in shIGF2BP2 chondrocytes at the transcriptome level. As shown in the volcano plots (Fig. 2A), we identified 1,816 differentially-expressed genes (DEGs), of which 801 were downregulated and 1,015 were upregulated (Fig. S2A, B). The top 10 significantly upregulated KEGG enrichment pathways in the shIGF2BP2 group versus the shNC group showed that IGF2BP2 deficiency was mainly related to inflammation pathways, such as the “NF-κB signaling pathway”, the “MAPK signaling pathway” and the “interleukin-17 signaling pathway” (Fig. 2B). Besides, a notable differential expression of ECM degradation related genes, namely ADAMTS1, ADAMTS9 and MMP9, was observed between shNC and shIGF2BP2 samples (Fig. 2C), which indicated that ECM degradation was fostered by IGF2BP2 knockdown.

Fig. 2.

Fig. 2

Open in a new tab

IGF2BP2 knockdown causes disturbance of ECM homeostasis and cell cycle transition. (A) Volcano plot of the differentially-expressed genes between the shIGF2BP2 group and the shNC group. (B) The top 10 significantly-upregulated KEGG enrichment pathways in the shIGF2BP2 group compared with the shNC group. (C) Log2FC-based relative mRNA expression heat map of factors enriched in the “Degradation of the extracellular matrix” related genes in the shIGF2BP2 group compared with those from the shNC group. (D) The results of GSEA pathway enrichment analysis after RNA-seq. (E) Relative expression of selected downstream targets (MMP3, MMP10, COL2A1, TP53, CDKN1A and CXCL3) in the shIGF2BP2 and the shNC chondrocytes. Expression levels of these genes were normalized to control. (F) Immunofluorescence staining of p53 (green) and p21 (green), DAPI (blue) applied as a nuclear marker. Data were analyzed using one-way ANOVA and are expressed as mean with 95%CI. KEGG, Kyoto Encyclopedia of Genes and Genomes

To further investigate the effect of IGF2BP2 deficiency at the transcriptional level, gene set enrichment analysis (GSEA) was performed (Fig. 2D). Several KEGG pathways were identified, including the “NF-κB signaling pathway”, the “TNF signaling pathway” and the “MAPK signaling pathway”, indicating the involvement of inflammatory signaling pathways. Notably, these data also tipped several senescence-related signaling pathways, including the “NF-κB signaling pathway” and the “Cell Cycle”. Based on our downstream target screening strategy and pathway enrichment analysis, key molecules associated with ECM, SASP and cellular senescence (Matrix Metalloproteinases 3 (MMP3), MMP10, Collagen Type II Alpha 1 Chain (COL2A1), Tumor Protein 53 (TP53), Cyclin Dependent Kinase Inhibitor 1 A (CDKN1A)) and C-X-C motif chemokine ligand 3 (CXCL3)) were selected for initial validation by using qRT-PCR. The shIGF2BP2 group exhibited significant downregulation of COL2A1 with concurrent upregulation of MMP3, MMP10, TP53, CDKN1A and CXCL3 (Fig. 2E).

Consequently, the effect of IGF2BP2 deficiency on cell cycle transition was investigated by immunofluorescent staining. As shown in Fig. 2F and Fig. S3A, compared with Control and shNC chondrocytes, p53 (a key cell cycle regulatory factor) was significantly increased in the nucleus instead of the cytoplasm following IGF2BP2 knockdown. Simultaneously, expression of p21 in the nucleus (a key downstream target of p53) was also found to be significantly upregulated, which was consistent with the result of qRT-PCR as CDKN1A was upregulated in the shIGF2BP2 group (Fig. S3B). WB analysis also showed increased protein levels of p53, p21 and p16 after IGF2BP2 knockdown in chondrocytes (Fig. S4). These findings confirmed that deficiency of IGF2BP2 may disturb cell cycle transition and promote senescence.

Loss of IGF2BP2 induces chondrocyte senescence

To comprehensively elucidate the underlying mechanisms through which IGF2BP2 regulates cartilage homeostasis, Tandem Mass Tag (TMT) proteomic analysis was applied to shNC and shIGF2BP2 chondrocytes. As shown in the volcano plots (Fig. 3A), we identified 534 differentially expressed proteins, of which 265 proteins were upregulated and 269 proteins were downregulated in shIGF2BP2 chondrocytes (Fig. S5A, B). From the significantly upregulated genes with altered expression, we focused on pathways related to cellular senescence and cell cycle arrest via Reactome enrichment analysis (including Inactivation of APC/C via direct inhibition of the APC/C complex, Regulation of TP53 activity through phosphorylation, Formation of Senescence-Associated Heterochromatin Foci (SAHF), Inhibition of the proteolytic activity of APC/C required for the onset of anaphase by mitotic spindle checkpoint components, and TP53 regulation of Transcription of Genes Involved in G2 Cell Cycle Arrest and SASP) (Fig. 3B). Furthermore, the heatmap revealed the expression of screened genes in the shIGF2BP2 and shNC groups (Fig. 3C): Ⅰ. BUB1 Mitotic Checkpoint Serine/Threonine Kinase B (BUB1B), Ubiquitin Conjugating Enzyme E2 C (UBE2C), UBE2S, and Cell Division Cycle 20 (CDC20) were highly expressed in the shIGF2BP2 group, suggesting loss of IGF2BP2 might influence mitotic spindle checkpoint [27, 28]. Ⅱ. Aurora Kinase A (AURKA), Cyclin B1 (CCNB1), Cyclin Dependent Kinase 1 (CDK1) and BCL2 Associated X (BAX) were associated with G2 cell cycle arrest and were highly expressed in the shIGF2BP2 group [29]. Ⅲ. High Mobility Group AT-Hook 1 (HMGA1) and HMGA2 exhibited a high level of expression, which was associated with formation of SAHF [30]. Ⅳ. Jun Proto-Oncogene (JUN), Cyclin Dependent Kinase 2 (CDK2), Anaphase Promoting Complex Subunit 2 (ANAPC2), Mitogen-Activated Protein Kinase 1 (MAPK1), Cell Division Cycle 23 (CDC23) and Nuclear Factor Kappa B Subunit 1 (NFKB1) were implicated in the SASP [31], and all were highly expressed in the shIGF2BP2 group.

Fig. 3.

Fig. 3

Open in a new tab

Loss of IGF2BP2 induces chondrocyte senescence. (A) Volcano plot of the differentially-expressed proteins between the shIGF2BP2 group and the shNC group. (B) Reactome enrichment analysis. (C) Heat map of clustering analysis. (D) Brightfield images of senescence-associated beta-galactosidase (SA-β-gal) staining. (E) Representative images of EdU immunofluorescence (green) with Hoechst 33324 staining applied as a nuclear counter-staining dye (blue) in the Control, shNC and shIGF2BP2 groups. (F) Representative images of γH2AX (green) immunofluorescence with DAPI staining applied as a nuclear counter-staining dye (blue) in the Control, shNC and shIGF2BP2 groups. (G) Flow cytometry analysis of cell cycle in Control, shNC and shIGF2BP2 chondrocytes. Data were analyzed using one-way ANOVA test

To further substantiate these results, the senescence-associated markers senescence-associated β-galactosidase (SA-β-gal), 5-ethynyl-2’-deoxyuridine (EdU) and γH2AX were analyzed. SA-β-gal activity is a key marker of cellular senescence [32]. SA-β-gal activity exhibited progressive elevation across experimental groups during the 4-day period. However, the increase in the intensity of SA-β-gal staining was much smaller in Control and shNC groups compared to that in the shIGF2BP2 group, suggesting that loss of IGF2BP2 might induce cellular senescence (Fig. 3D and Fig. S6). One of the defining characteristics of senescent cells is stable cell cycle arrest [33], therefore EdU staining was employed to detect lack of DNA replication and cell cycle arrest. EdU incorporation confirmed that a significant proportion of cells in Control and shNC groups were undergoing division, whereas most cells in the shIGF2BP2 group were arrested (Fig. 3E and Fig. S7). Moreover, labeling of γH2AX, a sensitive marker of DNA damage and a senescence-associated hallmark [34], was also higher in shIGF2BP2 cells compared with Control and shNC cells (Fig. 3F and Fig. S8). To properly discriminate senescent cells from cycling cells, we employed a well-established algorithm that evaluates two key markers: Ki67 to identify cycling cells; and SPiDER-βgal activity to identify senescent cells [35]. The results indicated that inhibition of IGF2BP2 increased the proportion of senescent cells and downregulated the proportion of cycling cells (Fig. S9). These findings suggested that loss of IGF2BP2 may increase DNA damage, thus contributing to senescence. Additionally, we employed flow cytometry to identify whether IGF2BP2 deficiency could affect cell cycle progression. The results indicated that inhibition of IGF2BP2 significantly increased the proportion of cells in G0/G1 phase and downregulated the proportion of cells in S phase (Fig. 3G).

Collectively, these results indicated that loss of IGF2BP2 may induce G1/S cell cycle arrest and cellular senescence.

Metabolic reprogramming is triggered by IGF2BP2 deficiency

In healthy cartilage, due to the hypoxic environment, chondrocytes are more likely to obtain energy by anaerobic glucose metabolism. Glycolysis, as the primary metabolic pattern of chondrocytes in healthy cartilage, plays an important role in maintaining normal function and preventing cartilage degeneration. Meanwhile, mitochondrial dysfunction has been reported to occur prior to cartilage degradation. However, the relationship between glycolysis, mitochondrial function and chondrocyte senescence remains unclear. Consequently, we examined the role of IGF2BP2 in glycolysis and mitochondrial function during cartilage degeneration.

To better understand the metabolic changes triggered by IGF2BP2 deficiency in chondrocytes, LC-MS/MS-based metabolomics combined with TMT proteomics were applied to the shNC and shIGF2BP2 C28/I2 cells. Among the 318 differential expressed metabolites, 59 metabolites were upregulated and 259 metabolites were downregulated in shIGF2BP2 chondrocytes (Fig. S10A, B).

As shown in Fig. 4A and B, KEGG enrichment analysis was conducted on the downregulated metabolites and proteins, revealing their predominant involvement in the metabolic processes of glycolysis. Heat map representation of the protein constituents of the respiratory chain showed a strong upregulation between shNC and shIGF2BP2 samples, revealing that upregulation of the protein components of mitochondrial complex I is the most pervasive and significant (Fig. 4C). Moreover, heat map representation of the key enzymes (including HK1, PGK2, ALDOB and ENO2) and metabolites (including fructose-1,6-biphosphate, glyceraldehyde-3-phosphate, pyruvate and lactate) revealed a positively-correlated regulation of the glycolysis process between shNC and shIGF2BP2 samples (Fig. 4D). The results revealed that key enzymes of glycolysis exhibited general decreases in expression while protein constituents of oxidative phosphorylation exhibited significant upregulation following IGF2BP2 knockdown.

Fig. 4.

Fig. 4

Open in a new tab

Metabolic reprogramming is triggered by IGF2BP2 deficiency. (A) The top 20 significantly-downregulated KEGG enrichment pathways of metabolomics in the shIGF2BP2 group versus the shNC group. (B) The top 20 significantly-downregulated KEGG enrichment pathways identified in TMT proteomics in the shIGF2BP2 group versus the shNC group. (C) Heat map of the protein components of the mitochondrial respiratory chain complex in the shIGF2BP2 group versus shNC group. (D) Heatmap of key enzymes and main metabolites in glycolysis in the shIGF2BP2 group versus shNC group. (E) Fluorescence microscopic images of JC-10 staining to measure mitochondrial membrane potential in the Control, shNC and shIGF2BP2 groups. The red fluorescence represents JC-10 aggregates and the green fluorescence represents JC-10 monomers. (F) Fluorescence images of intracellular ROS analysis with DCFH-DA in the Control, shNC and shIGF2BP2 groups

Mitochondrial dysfunction emerges as a hallmark of cellular senescence, characterized by pathological mitochondrial fragmentation and cytoplasmic vacuolization in aged chondrocytes. This dysfunction leads to heightened ROS production and reduced membrane potential (ΔΨm). The elevated ROS from mitochondrial dysfunction worsens the damage to normal mitochondria. Thus, JC-10 fluorescence analysis was used to monitor the ΔΨm. As shown in Fig. 4E and Fig. S11, JC-10 was mostly present in its monomer form, which emits green fluorescence, in shIGF2BP2 chondrocytes, while JC-10 aggregates, which emit red fluorescence, were the primary JC-10 form in Control and shNC chondrocytes. Mitochondria are the main source of cellular reactive oxygen species (ROS) [36], which play a vital role in the onset of cartilage degradation and OA progression [37]. Therefore, intracellular ROS generation in all groups of cells was measured using 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) as a fluorescent probe. As shown in Fig. 4F and Fig. S12, the intracellular ROS level was remarkably low in Control and shNC groups, but after knockdown of IGF2BP2, the intracellular ROS level exhibited a significant increase. These results indicated that mitochondrial dysfunction was triggered by loss of IGF2BP2.

HIF-1α preserves mitophagy and inhibits senescence induced by IGF2BP2 loss

Based on the observed metabolic shift triggered by IGF2BP2 deficiency and the significant downregulation of HIF-1 signaling pathway in proteomics (Fig. S13A, B), we hypothesized that lack of IGF2BP2 influenced cartilage chondrocytes through downregulating HIF-1α, a crucial transcriptional factor for glycometabolism and ECM homeostasis. In addition, correlation pattern analysis in cartilage tissue samples indicated that IGF2BP2 expression was positively linear with HIF-1α in the GSE114007 dataset (p = 0.0485, r = 0.323) (Fig. S13C).

To further validate this supposition, we investigated the expression of HIF-1α in shIGF2BP2 chondrocytes by immunofluorescence and WB. As shown in Fig. 5A, HIF-1α expression was decreased and relocated from the nucleus to the cytoplasm due to IGF2BP2 knockdown (Fig. S13D). Consistently, WB analysis showed decreased protein levels of HIF-1α after IGF2BP2 knockdown in chondrocytes (Fig. 5B and Fig. S14A).

Fig. 5.

Fig. 5

Open in a new tab

HIF-1α preserves mitophagy and inhibits senescence induced by IGF2BP2 loss. (A) Representative immunofluorescence images of HIF-1α in chondrocytes with shIGF2BP2 treatment. (B) Western blotting analysis of HIF-1α in chondrocytes of the shNC and shIGF2BP2 groups. n  =  3 per group. (C) Representative images of the MitoTracker™ Red fluorescence in the Control, shNC and shIGF2BP2 groups. (D) Western blotting analysis of PINK1 and Parkin in the shNC and shIGF2BP2 groups; n = 3 per group. (E-G) Rescue experiments were performed in C28/I2 chondrocytes to validate the relationship between IGF2BP2 and HIF-1α, accompanied by markers of ECM deposition, senescence and the PINK1/Parkin pathway. pCDNA3.1-HIF1A-3flag was transfected into shIGF2BP2 chondrocytes. After 48 h, the related genes were analyzed

As HIF-1α has been reported to act as a protector against OA progression by promoting mitophagy in OA cartilage [38], we then employed mitochondrial staining with MitoTracker™ Red to evaluate the morphology of mitochondria. In shIGF2BP2 chondrocytes, we observed that mitochondria exhibited decreased volume and increased sphericity, which contrasted with the normal rod-shape and regular arrangement of cristae detected in Control and shNC chondrocytes (Fig. 5C). Subsequently, the mitophagy markers PTEN Induced Kinase 1 (PINK1) and Parkin were assessed in shIGF2BP2 chondrocytes. As expected, PINK1 and Parkin were both downregulated due to loss of IGF2BP2 (Fig. 5D and Fig. S14B, C).

We then examined the effect of HIF-1α overexpression in shIGF2BP2 chondrocytes. As shown in Fig. 5E and F, IGF2BP2 loss induced decreases in ECM-related markers (COL Ⅱ, SOX9 and ACAN), downregulation of G1/S checkpoint-related proteins (CyclinD1, CDK4 and CDK6), and increases in expression of senescence markers (p53, p21 and p16) and SASP markers (MMP3, IL-1β and IL-6). The effects triggered by IGF2BP2 knockdown were partly reversed by HIF-1α overexpression (Fig. S15, 16).

Notably, reduced expression of PINK1 and Parkin were rescued by HIF-1α supplementation in IGF2BP2-deficient chondrocytes, indicating that HIF-1α may restore mitophagy impaired by IGF2BP2 loss (Fig. 5G and Fig. S17). Collectively, these results suggested that HIF-1α overexpression protected against chondrocyte senescence and ECM degradation caused by IGF2BP2 loss.

IGF2BP2 deficiency promotes inflammation and disrupts mismatch repair

To better understand cellular changes triggered by IGF2BP2 deficiency in chondrocytes, TMT proteomics combined with RNA-seq were applied to the C28/I2 cells transfected with shNC and shIGF2BP2. Combined analysis of the transcriptome and proteome indicated that there was a total of 268 common mapped pathways among 314 gene-related mapped pathways and 286 protein-related mapped pathways. As shown in the bubble chart of enriched KEGG pathways of common differentially-expressed substances (Fig. 6A), the IL-17 signaling pathway, cell cycle and glycolysis pathways were found to be enriched in both transcriptome and proteome. Notably, the pathway enrichment of glycolysis suggested that IGF2BP2 deficiency might disrupt the key enzymes of glycolysis at the transcriptional level. KEGG analysis revealed the involvement of several senescence-related pathways, including cell cycle and p53 signaling pathways, and several inflammatory signaling and inflammation factor signaling pathways, such as the MAPK signaling pathway, NF-κB signaling pathway, JAK-STAT signaling pathway, Toll-like receptor (TLR) signaling pathway and tumor necrosis factor (TNF) signaling pathway (Fig. 6B). Consistently, WB analysis showed that the protein level of NF-κB p65 was increased after knockdown of IGF2BP2 (Fig. S18).

Fig. 6.

Fig. 6

Open in a new tab

IGF2BP2 deficiency promotes inflammation and disrupts mismatch repair. (A) Bubble chart of common significantly-regulated pathways in both proteomics and transcriptomics. (B) Network showing the connections between enriched KEGG pathways that were common in both proteomics and transcriptomics. The size of the nodes reflect the term significance. (C) The results of GSEA pathway enrichment analysis of IL-17 signaling, Calcium signaling pathway and mismatch repair in both proteomics and transcriptomics. (D) The expression heatmap of key molecular constituents of the IL-17 signaling pathway and Mismatch Repair pathway in the shIGF2BP2 group versus shNC group

Further, integrated proteomics and transcriptomics GSEA revealed conserved pathway dysregulation in shIGF2BP2 cells, with marked activation of IL-17 signaling pathway and Calcium signaling pathway and suppression of mismatch repair across both omics layers (Fig. 6C). Heatmap analysis revealed coordinated expression patterns across functionally related genes and regulatory proteins associated with IL-17 signaling and mismatch repair pathways (including NFKB1, MAPK4, CASP8, ANAPC5, IKBKG, MSH2 and MSH3) exhibited the same regulation tendency in response to treatments with shNC and shIGF2BP2 (Fig. 6D).

Results of previous studies suggest that IL-17 signaling activates inflammatory transcription factors to induce gene expression via the transcription factor NF-κB and activation of the MAPK pathways [39, 40], which are in accord with our RNA-seq findings.

Mismatch repair (MMR), essential to correcting mistakes made by polymerases during DNA replication, loses efficiency with age, which accentuates the accumulation of genomic damage and the ectopic accumulation of DNA in the cytosol [41, 42].

Emerging roles of Calcium signaling pathway in OA progression has attracted attention [43]. For instance, abnormal activation of piezo type mechanosensitive ion channel (Piezo1), which causes Ca2+ flow into cells under stress stimulation, is reported to be closely associated with cartilage damage [44, 45]. Besides, inositol 1,4,5-trisphosphate receptor, type 2 (ITPR2) is also reported to promote cellular senescence through facilitating mitochondrial calcium accumulation and mitochondria–endoplasmic reticulum (ER) contacts [46].

Our data revealed that the inflammatory and senescence responses of chondrocytes under IGF2BP2 deficiency may be characterized by induction of the IL-17 signaling and Calcium signaling pathway and downregulated MMR mechanism.

Igf2bp2 deficiency promotes cartilage degeneration in vivo

As a degenerative joint disease, the pathological characteristics of OA include progressive loss of articular cartilage, osteophyte formation, synovial inflammation and remodeling of subchondral bone. Gross articular cartilage loss is the primary pathological feature of OA [47]. Articular chondrocytes are the only cell type responsible for producing ECM and sustaining the homeostasis of articular cartilage. As a model for an inducible chondrocyte-specific gene depletion, Col2a1-CreERT mice have been extensively used to investigate the functions of regulatory molecules governing the stability and integrity of articular cartilage [48]. To isolate cartilage-specific effects, we generated tamoxifen-inducible chondrocyte-targeted Igf2bp2 knockout mice (hereafter referred to as Col2a1-CreERT; Igf2bp2flox/flox mice) (Fig. S19A). Specifically, Igf2bp2flox/flox mice were first crossed with the Col2a1-CreERT strain to generate Col2a1-CreERT; Igf2bp2flox/+ mice. The Col2a1-CreERT; Igf2bp2flox/+ mice were then crossed with Igf2bp2flox/flox mice to obtain homozygous Col2a1-CreERT; Igf2bp2flox/flox mice (Fig. S19B). The genotypes were validated by PCR analyses (Fig. S19C).

Five successive doses of Tamoxifen were injected over one week, beginning once the mice reached 10-weeks-old, to induce Cre-mediated recombination in cartilage. Ten weeks after Tamoxifen injection, the femoral condyles of Wild Type mice, Igf2bp2flox/flox mice and Col2a1-CreERT; Igf2bp2flox/flox mice were harvested. Safranin O/Fast Green staining was employed to examine degradation in the cartilage of femoral condyles. We found clear layers of condylar cartilage structure, smooth and intact surface, and homogeneous distribution of proteoglycans in the articular surface of Wild Type and Igf2bp2flox/flox groups. By contrast, the Col2a1-CreERT; Igf2bp2flox/flox mice developed cartilage degeneration with burr-like changes, defective articular surfaces and reduced proteoglycan staining, leading to an elevated OARSI histological score (Fig. 7 and Fig. S20). Collectively, these results suggested that IGF2BP2 deficiency in chondrocytes may exacerbate cartilage degeneration.

Fig. 7.

Fig. 7

Open in a new tab

Igf2bp2 deficiency promotes cartilage degeneration in vivo. Representative images of Safranin O/Fast Green staining of femoral condyles from Igf2bp2flox/flox mice, Col2a1-CreERT;Igf2bp2flox/flox mice and Wild Type mice (n = 4 per group).

In conclusion, we present evidence suggesting that IGF2BP2 deficiency plays a role in OA cartilage degeneration by promoting senescence and metabolic reprogramming (Fig. 8). We identified that IGF2BP2 was significantly downregulated in osteoarthritis chondrocytes. Transcriptomics, proteomics and metabolomics analyses were used to investigate the potential regulatory mechanisms of IGF2BP2 on regulating cartilage homeostasis. In vitro, IGF2BP2 knockdown in chondrocytes exacerbated senescence and decreased ECM deposition, whereas supplementation of HIF-1α had a therapeutic effect, indicating the essential role of the IGF2BP2-HIF-1α axis in maintaining cartilage homeostasis. Moreover, IGF2BP2 deficiency impaired chondrocyte mitochondrial function through downregulating the mitophagy-related PINK1/Parkin pathway. Using Col2a1-CreERT; Igf2bp2flox/flox mice as an animal model, we found that IGF2BP2 deficiency in chondrocytes is a sufficient condition for the occurrence of cartilage degeneration.

Fig. 8.

Fig. 8

Open in a new tab

Schematic illustration of IGF2BP2 deficiency inducing chondrocyte senescence and cartilage degeneration by targeting mitochondrial function. Illustration credit: Lina Cao

Discussion

In this study, we focused on the function of the m6A reader IGF2BP2 in chondrocyte senescence and cartilage degeneration. We observed downregulation of IGF2BP2 in osteoarthritic chondrocytes, with parallel downregulation of HIF-1α - a key transcriptional regulator of mitochondrial metabolism. Mechanistically, knockdown of IGF2BP2 decreased the expression of HIF-1α, which further caused mitochondrial dysfunction and resulted in chondrocyte senescence. In addition, mice with chondrocyte-specific Igf2bp2-knockout (Col2a1-CreERT; Igf2bp2flox/flox) showed intensified cartilage degeneration in vivo.

Chondrocytes, the only resident cell type in mature cartilage, have a pivotal role in regulating both the catabolic and anabolic factors, maintaining a delicate balance between ECM synthesis and degradation [5]. Multiple factors, include aging, trauma, wear, and obesity, can disrupt the homeostasis of ECM anabolism and catabolism, which ultimately contribute to the progressive degeneration of cartilage due to its limited self-repair capacity [6]. Currently, few treatments possess the ability to reverse such an ECM imbalance [3]. Hence, it is essential to comprehend the mechanisms underlying ECM degeneration in cartilage to identify new therapeutic targets and develop effective drugs that slow the progression of cartilage degeneration. Our results showed a significant decrease in IGF2BP2 mRNA level in the cartilage tissues of OA patients. Results from the C28/I2 cell line indicated that IGF2BP2 deficiency leads to ECM degradation accompanied by senescence and mitochondrial dysfunction. These results suggest that downregulation of IGF2BP2 is an important factor leading to a series of downstream abnormalities in chondrocytes. Given the diversified biological role of IGF2BP2, exploration of its downstream mechanisms deserves further investigation in this study.

Senescence, one of the hallmarks of aging, is a cell state characterized by a prolonged and irreversible cell-cycle withdrawal with secretion of SASP and deregulated metabolism [7]. It is highly probable that the thread tying cellular senescence and OA pathology together is the accumulation of senescent chondrocytes coupled with gradual changes in cellular metabolism and function, all of which contribute to loss of cartilage homeostasis [10]. In this study, we observed cell cycle arrest and cellular senescence in shIGF2BP2 chondrocytes. As accumulating evidence has highlighted the correlation of IGF2BP2 inhibition with senescence and inflammation-related diseases [20, 21, 49, 50], it is worth investigating what underlies the loss of IGF2BP2 and its link with senescence. Regarding cellular senescence, previous study shows IGF2BP2 suppresses premature senescence of human bone marrow mesenchymal stem cells by stabilizing m6A-modified MIS12 mRNA, and cellular senescence exacerbates OA progression [20]. For deregulated metabolism and mitochondrial homeostasis, studies demonstrate IGF2BP2 depends on METTL14 to stabilize lncRNA TUG1 and regulate PGC1-α, a key regulator of mitochondrial function, while mitochondrial imbalance contributes to chondrocyte dysfunction in OA [51]. These previous studies may suggest the potential of investigating IGF2BP2’s role in OA through these targets.

Physiologically, chondrocytes exist at low oxygen tension, ranging from 10% at the surface to less than 1% in the deep zones. HIF-1α plays a key role in facilitating metabolic adaptation to the hypoxic environment and maintaining the homeostasis of articular cartilage [50]. Numerous researchers have reported that activation of HIF-1α maintains anaerobic glycolysis and mitochondrial function of chondrocytes via mitophagy [52] and regulates the chondrocyte ECM environment, resulting in promotion of ECM synthesis and delay of cartilage degradation [53]. In addition, emerging evidence also indicated that HIF-1α is involved in the regulation of senescence [54].

In our study, we first observed the downregulation of glycolysis and upregulation of OXPHOS protein level by metabolomics and proteomics, accompanied by decreased mitochondrial membrane potential and increased levels of ROS. We then identified downregulation of the HIF-1α signaling pathway in shIGF2BP2 chondrocytes, as revealed by KEGG analysis. As HIF-1α, considered to be the regulator of anaerobic glycolysis and mitochondrial function in chondrocytes, has also been reported to delay the onset of senescence, we proposed that HIF-1α could link senescence, mitochondrial dysfunction, and cartilage degeneration. We then analyzed the RNA-seq data of human OA chondrocytes (GSE114007) and found that the level of HIF-1α in chondrocytes was positively correlated with the level of IGF2BP2. The results of immunofluorescence and WB confirmed the absence of HIF-1α in shIGF2BP2 chondrocytes. Next, we demonstrated that overexpression of HIF-1α rescued the effect of IGF2BP2 deficiency on ECM degradation in chondrocytes, accompanied by downregulation of senescence-related biomarkers.

Moreover, the absence of IGF2BP2 compromises mitophagy by downregulating the expression of PINK1/Parkin. Parkin is an E3 ubiquitin ligase, which is essential for mitochondrial quality control in chondrocytes by recruiting depolarized mitochondria and removing impaired mitochondria. Studies have confirmed that mitophagy is related to senescence and serves as a therapeutic target for OA [55]. Our results showed a decrease in PINK1/Parkin expression in shIGF2BP2 chondrocytes at protein level and revealed that overexpression of HIF-1α reverses this downregulation.

Simultaneously, our GSEA analysis of differential expressed genes and proteins revealed a pronounced enrichment of inflammatory-associated pathways, specifically the IL-17 signaling pathway, in the shIGF2BP2 group. The IL-17 signaling pathway is involved in the maintenance of innate and adaptive immunity, and dysregulated production of IL-17 has been shown to promote OA development from multiple perspectives, promoting OA progression by regulating chondrocyte autophagy and senescence and cartilage matrix degradation, while inducing synovial inflammation by promoting the release of inflammatory cytokines and adipocytokines [56]. Our results suggest that inhibition of IGF2BP2 might promote activity of the IL-17 signaling pathway and induce senescence-related gene expression via the transcription factor NF-κB and activation of mitogen-activated protein kinase (MAPK) pathways [39]. In this study, we observed a reduction in the levels of mismatch repair, which suggested IGF2BP2 deficiency might accelerate the progression of cellular senescence by disturbing genome stability.

Age-dependent OA is associated with factors thought to contribute to cartilage degeneration, including reduced chondrocyte proliferation, increased apoptosis, and decreased production of ECM components [56]. Here, we constructed the Col2a1-CreERT; Igf2bp2flox/flox mouse model, which was associated with a significant expression of SASP and enhancement of ECM degradation, indicating that the Col2a1-CreERT; Igf2bp2flox/flox mouse model might mimic senescence-related OA.

While our multi-omics approach identified key pathways, the precise m6A-dependent targets of IGF2BP2 in chondrocytes remain undefined. The reliance on a single mouse model limited generalizability to other OA subtypes. Furthermore, the interplay between IGF2BP2-regulated mitophagy and IL-17 signaling warrants deeper investigation, as inflammatory crosstalk may amplify senescence phenotypes. Notably, a limitation of our multi-omics bioinformatics analysis stems from the relatively small sample size, which may compromise statistical power and restrict the generalizability of our findings. Moreover, the depth of cross-omics integrative analysis remains limited, with insufficient exploration of inter-layer molecular regulatory networks. Additionally, our interpretations rely partially on computational annotations, necessitating experimental validation to confirm causal relationships between the identified omics signatures.

In conclusion, our study reveals the essential role of IGF2BP2 as an m6A-binding protein in the pathogenesis of cartilage degeneration through downregulation of HIF-1α and extends the potential role of the PINK1/Parkin signaling pathway, which leads to the onset of senescence and mitochondrial dysfunction, in the progression of articular cartilage loss.

Resource availability

Lead contact

Further information and requests for resources and reagents should be directed to and will be fulfilled by the lead contact, Shang-Chun Guo (scguo@shsmu.edu.cn).

Materials availability

All related information or materials generated in this study are available upon reasonable request.

Data and code availability

The published article includes all data generated in this study.

Methods

All methods can be found in the accompanying Transparent Methods supplemental file.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary Material 1 (4.4MB, docx)
Supplementary Material 2 (324.3KB, csv)
Supplementary Material 3 (80.9KB, csv)

Acknowledgements

We extend our gratitude to the Animal Experimental Center of Shanghai Sixth People’s Hospital for assistance. We wish to acknowledge current and previous members of our laboratory. In addition, we thank Beyotime Biotechnology, Cyagen Biosciences, NewHelix Biotech, OE Biotech, Solarbio, and Servicebio for providing technical support. We sincerely thank Lina Cao for the design of the schematic diagram. Some illustrations were created with BioRender.com.

Authors’ contributions

All authors approved the final submitted manuscript. Zhi-Han Shen: Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization, Writing – Original Draft. Xu-Ran Li: Investigation, Methodology, Validation, Writing – Original Draft. Jian Lu: Resources, Writing – Original Draft. Po-Lin Liu: Data curation, Investigation, Methodology. Shu-Hang He: Investigation, Methodology, Validation. Qing-Song Deng: Investigation, Methodology, Validation. Ang Wang: Resources. Ren Zhang: Resources. Shi-Cong Tao: Conceptualization, Funding acquisition, Project administration, Supervision, Writing – Review & Editing. Shang-Chun Guo: Conceptualization, Funding acquisition, Supervision, Writing – Review & Editing.

Funding

The present study was supported by the National Natural Science Foundation of China (grant numbers 82072530, 82372566, and 82472595), Shanghai “Rising Stars of Medical Talent” Youth Development Program (Youth Medical Talents–Specialist Program), Shanghai Jiao Tong University School of Medicine “Two-Hundred Talent” Program (2022-017), Shanghai Sixth People’s Hospital Excellent Young Scientist Development Program (ynyq202101) and Shanghai Xuhui District Medical Research Project (grant numbers SHXH202322).

Declarations

Competing interests

Authors declare that they have no competing interests.

Footnotes

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Zhi-Han Shen, Xu-Ran Li and Jian Lu contributed equally to this work.

Contributor Information

Shi-Cong Tao, Email: sctao@shsmu.edu.cn.

Shang-Chun Guo, Email: scguo@shsmu.edu.cn.

References

  • 1.Steinmetz JD et al (2023) Global, regional, and national burden of osteoarthritis, 1990–2020 and projections to 2050: a systematic analysis for the Global Burden of Disease Study 2021. Lancet Rheumatol 5 e508–e522. 10.1016/S2665-9913(23)00163-7 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2.Yao Q et al (2023) Osteoarthritis: pathogenic signaling pathways and therapeutic targets. Signal Transduct Target Therapy 8:56. 10.1038/s41392-023-01330-w [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Bruno MC et al (2022) Injectable Drug Delivery Systems for Osteoarthritis and Rheumatoid Arthritis. ACS Nano 16:19665–19690. 10.1021/acsnano.2c06393 [DOI] [PubMed] [Google Scholar]
  • 4.Guo X et al (2023) Regeneration of articular cartilage defects: Therapeutic strategies and perspectives. J Tissue Eng 14:20417314231164765. 10.1177/20417314231164765 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Duan W et al (2021) Combination of chondrocytes and chondrons improves extracellular matrix production to promote the repairs of defective knee cartilage in rabbits. J Orthop Translation 28:47–54. 10.1016/j.jot.2021.01.004 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Michelacci YM, Baccarin RYA, Rodrigues NNP (2023) Chondrocyte Homeostasis and Differentiation: Transcriptional Control and Signaling in Healthy and Osteoarthritic Conditions. Life (Basel) 13. 10.3390/life13071460 [DOI] [PMC free article] [PubMed]
  • 7.Roelofs AJ, De Bari C (2023) Osteoarthritis year in review 2023: Biology. Osteoarthritis and Cartilage. 10.1016/j.joca.2023.11.002 [DOI] [PubMed]
  • 8.Gorgoulis V et al (2019) Cellular Senescence: Defining a Path Forward. Cell 179:813–827. 10.1016/j.cell.2019.10.005 [DOI] [PubMed] [Google Scholar]
  • 9.Watson E, Mikos AG (2023) Advances in In Vitro and In Vivo Bioreactor-Based Bone Generation for Craniofacial Tissue Engineering. BME Front 4:0004. 10.34133/bmef.0004 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Coryell PR, Diekman BO, Loeser RF (2021) Mechanisms and therapeutic implications of cellular senescence in osteoarthritis. Nat Rev Rheumatol 17:47–57. 10.1038/s41584-020-00533-7 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Faust HJ et al (2020) IL-17 and immunologically induced senescence regulate response to injury in osteoarthritis. J Clin Invest 130:5493–5507. 10.1172/jci134091 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Liu W et al (2023) FTO-mediated m6A demethylation of pri-miR-3591 alleviates osteoarthritis progression. Arthritis Res Therapy 25:53. 10.1186/s13075-023-03035-5 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Zhang C et al (2024) POP1 Facilitates Proliferation in Triple-Negative Breast Cancer via m6A-Dependent Degradation of CDKN1A mRNA. Res (Wash D C) 7:0472. 10.34133/research.0472 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Wu Z et al (2023) m(6)A epitranscriptomic regulation of tissue homeostasis during primate aging. Nat Aging 3:705–721. 10.1038/s43587-023-00393-2 [DOI] [PubMed] [Google Scholar]
  • 15.Ding M et al (2024) Changes in M6A methylation: A key factor in the vicious cycle of flora -gut aging. Ageing Res Rev 98:102351. 10.1016/j.arr.2024.102351 [DOI] [PubMed] [Google Scholar]
  • 16.Shen D et al (2022) Detailed resume of RNA m6A demethylases. Acta Pharm Sinica B 12:2193–2205. 10.1016/j.apsb.2022.01.003 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Oerum S, Meynier V, Catala M, Tisné C (2021) A comprehensive review of m6A/m6Am RNA methyltransferase structures. Nucleic Acids Res 49:7239–7255. 10.1093/nar/gkab378 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Zhen D et al (2020) m(6)A Reader: Epitranscriptome Target Prediction and Functional Characterization of N (6)-Methyladenosine (m(6)A) Readers. Front Cell Dev Biol 8:741. 10.3389/fcell.2020.00741 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Zhang Q et al (2024) N6-Methyladenosine in Cell-Fate Determination of BMSCs: From Mechanism to Applications. Res (Wash D C) 7:0340. 10.34133/research.0340 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Wu Z et al (2020) METTL3 counteracts premature aging via m6A-dependent stabilization of MIS12 mRNA. Nucleic Acids Res 48:11083–11096. 10.1093/nar/gkaa816 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Ma XX et al (2024) Dual Role of IGF2BP2 in Osteoimmunomodulation during Periodontitis. J Dent Res 103:208–217. 10.1177/00220345231216115 [DOI] [PubMed] [Google Scholar]
  • 22.Chen L, Liu J, Rao Z (2024) FTO-overexpressing extracellular vesicles from BM-MSCs reverse cellular senescence and aging to ameliorate osteoarthritis by modulating METTL3/YTHDF2-mediated RNA m6A modifications. Int J Biol Macromol 278:134600. 10.1016/j.ijbiomac.2024.134600 [DOI] [PubMed] [Google Scholar]
  • 23.Zeng Z et al (2023) The m6A reader YTHDF2 alleviates the inflammatory response by inhibiting IL-6R/JAK2/STAT1 pathway-mediated high-mobility group box-1 release. Burns Trauma 11:tkad023. 10.1093/burnst/tkad023 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Wan L et al (2022) Role of m6A modification and novel circ_0066715/ miR-486-5p/ ETS1 axis in rheumatoid arthritis macrophage polarization progression. Aging 14:10009–10026. 10.18632/aging.204439 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25.Nan Y et al (2024) IGF2BP2 regulates the inflammation of fibroblast-like synoviocytes via GSTM5 in rheumatoid arthritis. Cell Death Discov 10:215. 10.1038/s41420-024-01988-3 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Suo M et al (2022) Age-dependent effects of Igf2bp2 on gene regulation, function, and aging of hematopoietic stem cells in mice. Blood 139:2653–2665. 10.1182/blood.2021012197 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.McAinsh AD, Kops GJPL (2023) Principles and dynamics of spindle assembly checkpoint signalling. Nat Rev Mol Cell Biol 24:543–559. 10.1038/s41580-023-00593-z [DOI] [PubMed] [Google Scholar]
  • 28.Lara-Gonzalez P, Westhorpe FG, Taylor SS (2012) The spindle assembly checkpoint. Curr Biol 22:R966–980. 10.1016/j.cub.2012.10.006 [DOI] [PubMed] [Google Scholar]
  • 29.Kumari R, Jat P (2021) Mechanisms of Cellular Senescence: Cell Cycle Arrest and Senescence Associated Secretory Phenotype. Front Cell Dev Biol 9:645593. 10.3389/fcell.2021.645593 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Zhang R, Chen W, Adams PD (2007) Molecular Dissection of Formation of Senescence-Associated Heterochromatin Foci. Mol Cell Biol 27:2343–2358. 10.1128/MCB.02019-06 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Wang B, Han J, Elisseeff JH, Demaria M (2024) The senescence-associated secretory phenotype and its physiological and pathological implications. Nat Rev Mol Cell Biol. 10.1038/s41580-024-00727-x [DOI] [PubMed] [Google Scholar]
  • 32.Debacq-Chainiaux F, Erusalimsky JD, Campisi J, Toussaint O (2009) Protocols to detect senescence-associated beta-galactosidase (SA-βgal) activity, a biomarker of senescent cells in culture and in vivo. Nat Protoc 4:1798–1806. 10.1038/nprot.2009.191 [DOI] [PubMed] [Google Scholar]
  • 33.Hernandez-Segura A, Nehme J, Demaria M (2018) Hallmarks of Cellular Senescence. Trends Cell Biol 28:436–453. 10.1016/j.tcb.2018.02.001 [DOI] [PubMed] [Google Scholar]
  • 34.Shi D et al (2023) Photoactivatable senolysis with single-cell resolution delays aging. Nat Aging 3:297–312. 10.1038/s43587-023-00360-x [DOI] [PubMed] [Google Scholar]
  • 35.Alessio N et al (2021) Different Stages of Quiescence, Senescence, and Cell Stress Identified by Molecular Algorithm Based on the Expression of Ki67, RPS6, and Beta-Galactosidase Activity. Int J Mol Sci 22. 10.3390/ijms22063102 [DOI] [PMC free article] [PubMed]
  • 36.Zhao RZ, Jiang S, Zhang L, Yu ZB (2019) Mitochondrial electron transport chain, ROS generation and uncoupling (Review). Int J Mol Med 44:3–15. 10.3892/ijmm.2019.4188 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37.Mobasheri A et al (2017) The role of metabolism in the pathogenesis of osteoarthritis. Nat Rev Rheumatol 13:302–311. 10.1038/nrrheum.2017.50 [DOI] [PubMed] [Google Scholar]
  • 38.Hu S et al (2020) Stabilization of HIF-1α alleviates osteoarthritis via enhancing mitophagy. Cell Death Dis 11:481. 10.1038/s41419-020-2680-0 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Li X, Bechara R, Zhao J, McGeachy MJ, Gaffen SL (2019) IL-17 receptor–based signaling and implications for disease. Nat Immunol 20:1594–1602. 10.1038/s41590-019-0514-y [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Amatya N, Garg AV, Gaffen SL (2017) IL-17 Signaling: The Yin and the Yang. Trends Immunol 38:310–322. 10.1016/j.it.2017.01.006 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41.López-Otín C, Blasco MA, Partridge L, Serrano M, Kroemer G (2023) Hallmarks of aging: An expanding universe. Cell 186:243–278. 10.1016/j.cell.2022.11.001 [DOI] [PubMed] [Google Scholar]
  • 42.Rona G et al (2024) CDK-independent role of D-type cyclins in regulating DNA mismatch repair. Mol Cell 84:1224–1242e1213. 10.1016/j.molcel.2024.02.010 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Zhou R, Fu W, Vasylyev D, Waxman SG, Liu CJ (2024) Ion channels in osteoarthritis: emerging roles and potential targets. Nat Rev Rheumatol 20:545–564. 10.1038/s41584-024-01146-0 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44.Gan D et al (2024) Piezo1 activation accelerates osteoarthritis progression and the targeted therapy effect of artemisinin. J Adv Res 62:105–117. 10.1016/j.jare.2023.09.040 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 45.Wang S et al (2022) Mechanical overloading induces GPX4-regulated chondrocyte ferroptosis in osteoarthritis via Piezo1 channel facilitated calcium influx. J Adv Res 41:63–75. 10.1016/j.jare.2022.01.004 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 46.Ziegler DV et al (2021) Calcium channel ITPR2 and mitochondria-ER contacts promote cellular senescence and aging. Nat Commun 12:720. 10.1038/s41467-021-20993-z [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47.He Y et al (2020) Pathogenesis of Osteoarthritis: Risk Factors, Regulatory Pathways in Chondrocytes, and Experimental Models. Biology (Basel) 9. 10.3390/biology9080194 [DOI] [PMC free article] [PubMed]
  • 48.Chen M, Li S, Xie W, Wang B, Chen D (2014) Col2CreER(T2), a mouse model for a chondrocyte-specific and inducible gene deletion. Eur Cell Mater 28:236–245. 10.22203/ecm.v028a16 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 49.Zhao Y et al (2024) IGF2BP2-Shox2 axis regulates hippocampal-neuronal senescence to alleviate microgravity-induced recognition disturbance. iScience 27:109917. 10.1016/j.isci.2024.109917 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 50.Wang X et al (2021) The m6A Reader IGF2BP2 Regulates Macrophage Phenotypic Activation and Inflammatory Diseases by Stabilizing TSC1 and PPARγ. Adv Sci (Weinh) 8:2100209. 10.1002/advs.202100209 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 51.Zhu Y et al (2024) M6A RNA Methylation-Mediated TUG1 Stability Maintains Mitochondrial Homeostasis during Kidney Aging by Epigenetically Regulating PGC1-α Expression. Antioxid Redox Signal 41:993–1013. 10.1089/ars.2024.0631 [DOI] [PubMed] [Google Scholar]
  • 52.Zeng CY, Wang XF, Hua FZ (2022) HIF-1α in Osteoarthritis: From Pathogenesis to Therapeutic Implications. Front Pharmacol 13:927126. 10.3389/fphar.2022.927126 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 53.Hu S et al (2020) Stabilization of HIF-1α alleviates osteoarthritis via enhancing mitophagy. Cell Death Dis 11:481. 10.1038/s41419-020-2680-0 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 54.Zheng B et al (2024) Simultaneously Modulating HIF-1α and HIF-2α and Optimizing Macrophage Polarization through the Biomimetic Gene Vector toward the Treatment of Osteoarthritis. Biomater Res 28:0059. 10.34133/bmr.0059 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 55.Gao H et al (2023) Role of hypoxia in cellular senescence. Pharmacol Res 194:106841. 10.1016/j.phrs.2023.106841 [DOI] [PubMed] [Google Scholar]
  • 56.Fang G et al (2023) FUNDC1/PFKP-mediated mitophagy induced by KD025 ameliorates cartilage degeneration in osteoarthritis. Mol Ther 31:3594–3612. 10.1016/j.ymthe.2023.10.016 [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supplementary Material 1 (4.4MB, docx)
Supplementary Material 2 (324.3KB, csv)
Supplementary Material 3 (80.9KB, csv)

Data Availability Statement

The published article includes all data generated in this study.

Articles from Cellular and Molecular Life Sciences: CMLS are provided here courtesy of Springer

RESOURCES