Epigenetic regulatory mechanism of ADAMTS12 expression in osteoarthritis

  • Mol Med. 2023 Jul 3;29(1):86. doi: 10.1186/s10020-023-00661-2.
Shu Yang  1 ,  Xuanping Zhou  2 ,  Zhen Jia  1 ,  Mali Zhang  1 ,  Minghao Yuan  1 ,  Yizhao Zhou  1 ,  Jing Wang  3 ,  Duo Xia  4
Affiliations
  • 1. Department of Orthopedics, Hunan Provincial People's Hospital (The First-affiliated Hospital of Hunan Normal University), No. 61, Jiefang West Road, Furong District, Changsha, Hunan, 410005, People's Republic of China.
  • 2. Department of Orthopedics, The First-affiliated Hospital of Hunan Normal University (Hunan Provincial People's Hospital), Changsha, Hunan, 410005, People's Republic of China.
  • 3. Department of Orthopedics, Hunan Provincial People's Hospital (The First-affiliated Hospital of Hunan Normal University), No. 61, Jiefang West Road, Furong District, Changsha, Hunan, 410005, People's Republic of China. [email protected].
  • 4. Department of Orthopedics, Hunan Provincial People's Hospital (The First-affiliated Hospital of Hunan Normal University), No. 61, Jiefang West Road, Furong District, Changsha, Hunan, 410005, People's Republic of China. [email protected].
PMID: 37400752 DOI: 10.1186/s10020-023-00661-2
Abstract

Background: Osteoarthritis (OA) is a degenerative joint disease with lacking effective prevention targets. A disintegrin and metalloproteinase with thrombospondin motifs 12 (ADAMTS12) is a member of the ADAMTS family and is upregulated in OA pathologic tissues with no fully understood molecular mechanisms.

Methods: The anterior cruciate ligament transection (ACL-T) method was used to establish rat OA models, and interleukin-1 beta (IL-1β) was administered to induce rat chondrocyte inflammation. Cartilage damage was analyzed via hematoxylin-eosin, Periodic Acid-Schiff, safranin O-fast green, Osteoarthritis Research Society International score, and micro-computed tomography assays. Chondrocyte Apoptosis was detected by flow cytometry and TdT dUTP nick-end labeling. Signal transducer and activator of transcription 1 (STAT1), ADAMTS12, and methyltransferase-like 3 (METTL3) levels were detected by immunohistochemistry, quantitative polymerase chain reaction (qPCR), western blot, or immunofluorescence assay. The binding ability was confirmed by chromatin immunoprecipitation-qPCR, electromobility shift assay, dual-luciferase reporter, or RNA immunoprecipitation (RIP) assay. The methylation level of STAT1 was analyzed by MeRIP-qPCR assay. STAT1 stability was investigated by actinomycin D assay.

Results: The STAT1 and ADAMTS12 expressions were significantly increased in the human and rat samples of cartilage injury, as well as in IL-1β-treated rat chondrocytes. STAT1 is bound to the promoter region of ADAMTS12 to activate its transcription. METTL3/ Insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) mediated N6-methyladenosine modification of STAT1 promoted STAT1 mRNA stability, resulting in increased expression. ADAMTS12 expression was reduced and the IL-1β-induced inflammatory chondrocyte injury was attenuated by silencing METTL3. Additionally, knocking down METTL3 in ACL-T-produced OA rats reduced ADAMTS12 expression in their cartilage tissues, thereby alleviating cartilage damage.

Conclusion: METTL3/IGF2BP2 axis increases STAT1 stability and expression to promote OA progression by up-regulating ADAMTS12 expression.

Keywords
ADAMTS12; IGF2BP2; Inflammation; METTL3; Osteoarthritis; STAT1.
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