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Kdm6b regulates cartilage development and homeostasis through anabolic metabolism
  1. Jun Dai1,2,
  2. Dongsheng Yu1,2,3,
  3. Yafei Wang1,2,
  4. Yishan Chen1,2,
  5. Heng Sun1,2,
  6. Xiaolei Zhang1,2,
  7. Shouan Zhu1,2,
  8. Zongyou Pan1,2,
  9. Boon Chin Heng4,
  10. Shufang Zhang1,2,
  11. Hongwei Ouyang1,2,3,5,6
  1. 1 Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Zhejiang, China
  2. 2 Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, School of Medicine, Zhejiang University, Zhejiang, China
  3. 3 Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China
  4. 4 Endodontology, Faculty of Dentistry, The University of Hong Kong, Pokfulam, Hong Kong
  5. 5 State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China
  6. 6 China Orthopedic Regenerative Medicine Group, Hangzhou, China
  1. Correspondence to Professor Hongwei Ouyang, Dr Li Dak Sum & Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang Provincial, Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, Zhejiang University, 866 Yu Hang Tang Road, Hangzhou 310000, China; hwoy{at}zju.edu.cn

Abstract

Objectives Epigenetic mechanisms have been reported to play key roles in chondrogenesis and osteoarthritis (OA) development. Here, we sought to identify specific histone demethylases that are involved and delineate the underlying mechanisms.

Methods We screened the expression of 17 distinct histone demethylases by quantitative real time PCR (qRT-PCR) during chondrogenic differentiation of C3H10T1/2 cells. The role of Kdm6b in cartilage development was then analysed with transgenic Col2a1-CreERT2;Kdm6bf/f . RNA-Seq was applied to explore the underlying changes in chondrocytes upon knockdown of Kdm6b. Experimental OA in mice was induced by destabilisation of the medial meniscus in C57BL/6J (wild type, Kdm6bf/f and Col2a1-CreERT2;Kdm6bf/f ) mice, either with intra-articular injection of shKdm6b lentivirus or after tamoxifen treatment. Mouse joints and human cartilage samples were used for histological analysis.

Results Kdm6b expression was significantly increased during cartilage development. Col2a1-CreERT2;Kdm6bf/f mice displayed obvious skeletal abnormalities at E16.5 and E18.5 with intraperitoneal injection of tamoxifen at E12.5. RNA-Seq and qRT-PCR analyses revealed decreased expression of chondrocyte anabolic genes in Col2a1-CreERT2;Kdm6bf/f chondrocytes. The histological OA score was significantly higher in mice injected with Kdm6b short hairpin RNA lentivirus. Col2a1-CreERT2;Kdm6bf/f mice exhibited accelerated OA development at 8 and 12 weeks following surgical induction. The number of Kdm6b-positive chondrocytes was lower in both mice and human OA cartilage samples.

Conclusions These findings indicate that knockdown of Kdm6b in chondrocytes leads to abnormal cartilage development and accelerated OA progression via inhibition of the anabolic metabolism of chondrocytes. Understanding the epigenetic mechanism of joint cartilage development and homeostasis would be useful for development of new therapeutic modalities for OA.

  • Chondrocytes
  • Knee Osteoarthritis
  • Osteoarthritis

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Footnotes

  • Handling editor Tore K Kvien

  • JD and DY contributed equally

  • Contributors JD, DY, SZhu and HO designed research; JD, DY, YW and YC performed research; JD, DY, YW, HS, XZ, ZP, SZha and HO analysed data; JD, DY, BCH and HO wrote the paper.

  • Funding This work was supported by the National Key Research and Development Program of China (2016YFB0700804), NSFC grants (81630065, GZ1094, 81472115) and the key scientific and technological innovation team of Zhejiang Province (2013TD11).

  • Competing interests None declared.

  • Patient consent Obtained.

  • Ethics approval Zhejiang University Ethics Committee.

  • Provenance and peer review Not commissioned; externally peer reviewed.

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