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METTL3 Regulates the m6A Modification of NEK7 to Inhibit the Formation of Osteoarthritis
Xiaochuan Xiong1, Hao Xiong1, Jun Peng1
1Department of Orthopaedics, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai, China.
Objective:
Osteoarthritis (OA) is a common degenerative joint disease. The occurrence of OA slowly destroys the soft tissue structure of the patient's joint. Severe cases could lead to disability. Current studies had shown that inhibition of chondrocytes pyroptosis could slow down the progression of OA. Our work aimed to explore the specific mechanisms and ways of regulating this process.
Design:
In this work, the level of N6-methyladenosine (m6A) in clinical tissues was detected by ribonucleic acid (RNA) m6A dot blot. qRT-PCR (quantitative real-time polymerase chain reaction) was used to detect the messenger RNA (mRNA) expression level of m6A modified enzyme in clinical tissues. MTT (3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromid) and flow cytometry were used to detect the effect of sh-METTL3 (methyltransferase like 3) and NIMA-related kinase 7 (NEK7) transfection on chondrocytes pyroptosis in OA. Western blot was used to detect the protein expression levels of pyroptosis-related proteins. ELISA (enzyme-linked immunosorbent assay) was used to measure the protein concentration of inflammatory cytokines. The SRAMP online database was used to predict the m6A site of NEK7. HE staining was used to assess the progression of OA in mice.
Results:
The level of m6A in clinical samples of OA patients was higher, and METTL3 was significantly higher expressed in clinical samples of OA patients. We provided evidence that low expression of METTL3 inhibited chondrocytes pyroptosis. In addition, Rescue experiments and in vivo experiments had shown that METTL3 in combination with NEK7 inhibited the progression of OA by promoting chondrocytes pyroptosis.
Conclusions:
METTL3 regulates m6A modification of NEK7 and inhibits OA progression.
Insights
This study reveals that higher N6-methyladenosine (m6A) levels and METTL3 expression are linked to osteoarthritis (OA). METTL3, by modifying NEK7, inhibits chondrocyte pyroptosis, thereby slowing OA progression.
Area of Science:
- Biochemistry
- Molecular Biology
- Pathology
Background:
- Osteoarthritis (OA) is a degenerative joint disease characterized by progressive destruction of joint soft tissues, potentially leading to disability.
- Inhibiting chondrocytes pyroptosis has emerged as a promising strategy to slow OA progression.
- Understanding the regulatory mechanisms of chondrocytes pyroptosis is crucial for developing effective OA treatments.
Purpose of the Study:
- To investigate the role of N6-methyladenosine (m6A) modification in osteoarthritis (OA).
- To explore the specific mechanisms by which METTL3 and NEK7 influence chondrocytes pyroptosis in OA.
- To identify potential therapeutic targets for OA by regulating chondrocytes pyroptosis.
Main Methods:
- Detection of m6A levels and METTL3 expression in clinical OA tissues using RNA m6A dot blot and qRT-PCR.
- Assessment of chondrocytes pyroptosis using MTT assays and flow cytometry following sh-METTL3 and NEK7 transfection.
- Western blot analysis of pyroptosis-related proteins and ELISA for inflammatory cytokines.
- Prediction of m6A sites on NEK7 using the SRAMP database and evaluation of OA progression in mice via HE staining.
Main Results:
- Elevated m6A levels and significantly higher METTL3 expression were observed in clinical OA samples.
- Low expression of METTL3 was found to inhibit chondrocytes pyroptosis.
- Rescue and in vivo experiments demonstrated that METTL3, in conjunction with NEK7, inhibits OA progression by promoting chondrocytes pyroptosis.
Conclusions:
- METTL3 plays a critical role in regulating m6A modification.
- METTL3-mediated m6A modification of NEK7 inhibits chondrocytes pyroptosis and consequently slows OA progression.
- Targeting the METTL3-NEK7 pathway presents a potential therapeutic strategy for osteoarthritis.
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