Single-cell RNA sequencing reveals cellular and molecular heterogeneity in fibrocartilaginous enthesis formation

  1. Tao Zhang
  2. Liyang Wan
  3. Han Xiao
  4. Linfeng Wang
  5. Jianzhong Hu
  6. Hongbin Lu  Is a corresponding author
  1. Xiangya Hospital Central South University, China

Abstract

The attachment site of the rotator cuff (RC) is a classic fibrocartilaginous enthesis, which is the junction between bone and tendon with typical characteristics of a fibrocartilage transition zone. Enthesis development has historically been studied with lineage tracing of individual genes selected a priori, which does not allow for the determination of single-cell landscapes yielding mature cell types and tissues. Here, in together with open source GSE182997 datasets (3 sample) provided by Fang et al, we applied Single-cell RNA sequencing (scRNA-seq) to delineate the comprehensive postnatal RC enthesis growth and the temporal atlas from as early as postnatal day 1 up to postnatal week 8. And we furtherly performed single cell spatial transcriptomic sequencing on postnatal day 1 mice enthesis, in order to deconvoluted bone-tendon junction (BTJ) chondrocytes onto spatial spots. In summary, we deciphered the cellular heterogeneity and the molecular dynamics during fibrocartilage differentiation. Combined with current spatial transcriptomic data, our results provide a transcriptional resource that will support future investigations of enthesis development at the mechanistic level and may shed light on the strategies for enhanced RC healing outcomes.

Data availability

All single-cell datasets created during this study are publicly available at the Gene Expression Omnibus (GSE223751). Any additional information required to re-analyze the data in the paper is available from the corresponding author upon request.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Tao Zhang

    Department of Sports Medicine, Xiangya Hospital Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Liyang Wan

    Department of Sports Medicine, Xiangya Hospital Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2194-1080
  3. Han Xiao

    Department of Sports Medicine, Xiangya Hospital Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Linfeng Wang

    Department of Sports Medicine, Xiangya Hospital Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Jianzhong Hu

    Department of Sports Medicine, Xiangya Hospital Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.
  6. Hongbin Lu

    Department of Sports Medicine, Xiangya Hospital Central South University, Changsha, China
    For correspondence
    hongbinlu@hotmail.com
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7749-3593

Funding

National Natural Science Foundation of China (82230085)

  • Hongbin Lu

National Natural Science Foundation of China (82272572)

  • Hongbin Lu

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Murim Choi, Seoul National University, Republic of Korea

Ethics

Animal experimentation: All animal experimental protocols were approved by the Animal Ethics Committee of Central South University (No. 2022020058).

Version history

  1. Received: December 30, 2022
  2. Preprint posted: February 3, 2023 (view preprint)
  3. Accepted: September 10, 2023
  4. Accepted Manuscript published: September 12, 2023 (version 1)
  5. Version of Record published: September 21, 2023 (version 2)

Copyright

© 2023, Zhang et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Tao Zhang
  2. Liyang Wan
  3. Han Xiao
  4. Linfeng Wang
  5. Jianzhong Hu
  6. Hongbin Lu
(2023)
Single-cell RNA sequencing reveals cellular and molecular heterogeneity in fibrocartilaginous enthesis formation
eLife 12:e85873.
https://doi.org/10.7554/eLife.85873

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https://doi.org/10.7554/eLife.85873

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