Article Text

Single-cell RNA-seq analysis identifies meniscus progenitors and reveals the progression of meniscus degeneration
  1. Hao Sun1,2,
  2. Xingzhao Wen2,
  3. Hongyi Li2,
  4. Peihui Wu2,
  5. Minghui Gu2,
  6. Xiaoyi Zhao2,
  7. Ziji Zhang2,
  8. Shu Hu2,
  9. Guping Mao2,
  10. Ruofan Ma1,
  11. Weiming Liao2,
  12. Zhiqi Zhang2
  1. 1 Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Guangzhou, China
  2. 2 Department of Joint Surgery, Sun Yat-sen University First Affiliated Hospital, Guangzhou, China
  1. Correspondence to Dr Zhiqi Zhang, Department of Joint Surgery, Sun Yat-sen University First Affiliated Hospital, Guangzhou 510080, China; zhzhiqi{at}mail.sysu.edu.cn; Dr Weiming Liao; liaowmsysu{at}163.com

Abstract

Objectives The heterogeneity of meniscus cells and the mechanism of meniscus degeneration is not well understood. Here, single-cell RNA sequencing (scRNA-seq) was used to identify various meniscus cell subsets and investigate the mechanism of meniscus degeneration.

Methods scRNA-seq was used to identify cell subsets and their gene signatures in healthy human and degenerated meniscus cells to determine their differentiation relationships and characterise the diversity within specific cell types. Colony-forming, multi-differentiation assays and a mice meniscus injury model were used to identify meniscus progenitor cells. We investigated the role of degenerated meniscus progenitor (DegP) cell clusters during meniscus degeneration using computational analysis and experimental verification.

Results We identified seven clusters in healthy human meniscus, including five empirically defined populations and two novel populations. Pseudotime analysis showed endothelial cells and fibrochondrocyte progenitors (FCP) existed at the pseudospace trajectory start. Melanoma cell adhesion molecule ((MCAM)/CD146) was highly expressed in two clusters. CD146+ meniscus cells differentiated into osteoblasts and adipocytes and formed colonies. We identified changes in the proportions of degenerated meniscus cell clusters and found a cluster specific to degenerative meniscus with progenitor cell characteristics. The reconstruction of four progenitor cell clusters indicated that FCP differentiation into DegP was an aberrant process. Interleukin 1β stimulation in healthy human meniscus cells increased CD318+ cells, while TGFβ1 attenuated the increase in CD318+ cells in degenerated meniscus cells.

Conclusions The identification of meniscus progenitor cells provided new insights into cell-based meniscus tissue engineering, demonstrating a novel mechanism of meniscus degeneration, which contributes to the development of a novel therapeutic strategy.

  • cytokines
  • arthritis
  • fibroblasts
https://creativecommons.org/licenses/by/4.0/

This is an open access article distributed in accordance with the Creative Commons Attribution 4.0 Unported (CC BY 4.0) license, which permits others to copy, redistribute, remix, transform and build upon this work for any purpose, provided the original work is properly cited, a link to the licence is given, and indication of whether changes were made. See: https://creativecommons.org/licenses/by/4.0/.

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Footnotes

  • Handling editor Josef S Smolen

  • HS and XW contributed equally.

  • Contributors All listed authors meet the criteria for authorship and have contributed to the study design, data generation, data analysis, manuscript writing and manuscript review.

  • Funding This study was funded by the National Natural Science Foundation of China (no. 81874016, 81672145, 81472101, 81572119) and the Science and Technology Project of Guangzhou City, China (201710010164).

  • Competing interests None declared.

  • Patient consent for publication Not required.

  • Ethics approval Protocol approved by the Ethical Committee of The First Affiliated Hospital of Sun Yat‐sen University.

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

  • Data availability statement Data are available in a public, open access repository. The single-cell RNA-seq data, quality control information and cluster information are available at the NCBI’s Gene Expression Omnibus (GEO) data repository with the accession ID GSE133449.

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