Single-Cell RNA Sequencing Reveals Transcriptional Changes in the Cartilage of Subchondral Insufficiency Fracture of the Knee

J Inflamm Res. 2022 Nov 5:15:6105-6112. doi: 10.2147/JIR.S385648. eCollection 2022.

Abstract

Purpose: Subchondral insufficiency fracture of the knee (SIFK) is a common cause of knee joint pain that mainly afflicts the elderly. Until now, how a sudden insufficiency fracture of subchondral bone affects the transcriptomic profiles of cartilage in SIFK and OA patients are largely unknown.

Methods: Single-cell RNA sequencing (scRNA-seq) was used to identify various cell subsets and evaluate transcriptomic differences in cartilage of SIFK and OA patients. In addition, the above findings were confirmed by histological evaluation and immunohistochemical (IHC) staining.

Results: We found that the transcriptomic profiles of cartilage in the SIFK patient was completely different from those of normal and OA patients. Accordingly, several novel cell clusters with activation of hypoxia and endochondral ossification signaling were identified in the SIFK cartilage. Chondrocyte trajectories analysis and IHC staining revealed that transcription factors including TCF4 were found to be highly up-regulated during the occurrence of SIFK, which might drive the reactive formation of cartilage and fibrous tissue and the activation of endochondral ossification.

Conclusion: This is the first report to elucidate the transcriptomic alterations and distinct cell type subpopulations in the cartilage of SIFK and OA by the use of scRNA-seq, which provides a new insight in the understanding of the initiation and progression of SIFK.

Keywords: SIFK; TCF4; cartilage; hypoxia; scRNA.

Grants and funding

This work was supported by the National Key R&D Program of China (2021YFA1102600), the National Natural Science Foundation of China (82072470, 81871809, 81602360, 81672224, 82103781), the Natural Science Foundation of Guangdong Province (2021A1515012154, 2019A1515011082, 2017A030313665), the Guangzhou Science and Technology Project (202102010069, 201707010493), and the Macau Foundation for Development of Science and Technology (0029/2019/A).