1. Stem Cells and Regenerative Medicine

Mechanical stimulation promotes enthesis injury repair by mobilizing Prrx1+ cells via ciliary TGF-β signaling

  1. Han Xiao
  2. Tao Zhang
  3. Chang Jun Li
  4. Yong Cao
  5. Lin Feng Wang
  6. Hua Bin Chen
  7. Sheng Can Li
  8. Chang Biao Guan
  9. Jian Zhong Hu
  10. Di Chen
  11. Can Chen  Is a corresponding author
  12. Hong Bin Lu  Is a corresponding author
  1. Xiangya Hospital Central South University, China
  2. Chinese Academy of Sciences, China
https://doi.org/10.7554/eLife.73614
Share this article
Cite this article
  1. Han Xiao
  2. Tao Zhang
  3. Chang Jun Li
  4. Yong Cao
  5. Lin Feng Wang
  6. Hua Bin Chen
  7. Sheng Can Li
  8. Chang Biao Guan
  9. Jian Zhong Hu
  10. Di Chen
  11. Can Chen
  12. Hong Bin Lu
(2022)
Mechanical stimulation promotes enthesis injury repair by mobilizing Prrx1+ cells via ciliary TGF-β signaling
eLife 11:e73614.
https://doi.org/10.7554/eLife.73614
  2. Download BibTeX
  3. Download .RIS

Abstract

Proper mechanical stimulation can improve rotator cuff enthesis injury repair. However, the underlying mechanism of mechanical stimulation promoting injury repair is still unknown. In this study, we found that Prrx1+ cell was essential for murine rotator cuff enthesis development identified by single-cell RNA sequence and involved in the injury repair. Proper mechanical stimulation could promote the migration of Prrx1+ cells to enhance enthesis injury repair. Meantime, TGF-β signaling and primary cilia played an essential role in mediating mechanical stimulation signaling transmission. Proper mechanical stimulation enhanced the release of active TGF-β1 to promote migration of Prrx1+ cells. Inhibition of TGF-β signaling eliminated the stimulatory effect of mechanical stimulation on Prrx1+ cell migration and enthesis injury repair. In addition, knockdown of Pallidin to inhibit TGF-βR2 translocation to the primary cilia or deletion of Ift88 in Prrx1+ cells also restrained the mechanics-induced Prrx1+ cells migration. These findings suggested that mechanical stimulation could increase the release of active TGF-β1 and enhance the mobilization of Prrx1+ cells to promote enthesis injury repair via ciliary TGF-β signaling.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for Figures 1 and 2. Figure 3 - Source Data, Figure 4 - Source Data, Figure 5 - Source Data, Figure 6 - Source, Figure 7 - Source Data, Figure 8 - Source Data contain the numerical data used to generate the figures.

Article and author information

Author details

  1. Han Xiao

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

  2. Tao Zhang

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

  3. Chang Jun Li

    Department of Endocrinology, Xiangya Hospital Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.

  4. Yong Cao

    Department of Spine Surgery, Xiangya Hospital Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.

  5. Lin Feng Wang

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

  6. Hua Bin Chen

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

  7. Sheng Can Li

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

  8. Chang Biao Guan

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

  9. Jian Zhong Hu

    Department of Spine Surgery, Xiangya Hospital Central South University, Changsha, China
    Competing interests
    The authors declare that no competing interests exist.

  10. Di Chen

    Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China
    Competing interests
    The authors declare that no competing interests exist.

  11. Can Chen

    Department of Orthopedic, Xiangya Hospital Central South University, Changsha, China
    For correspondence
    chencan@csu.edu.cn
    Competing interests
    The authors declare that no competing interests exist.

  12. Hong Bin 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 (No. 81730068)

  • Hong Bin Lu

Major Science and technology progect of Changsha Science and Technology Bureau (No. 41965)

  • Hong Bin 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. Fayez Safadi, Northeast Ohio Medical University, United States

Ethics

Animal experimentation: All animal care protocols and experiments in this study were reviewed and approved by the Animal Care and Use Committees of the Laboratory Animal Research Center of our institute. All mice were maintained in the specific pathogen-free facility of the Laboratory Animal Research Center.

Version history

  1. Received: September 5, 2021
  2. Preprint posted: September 22, 2021 (view preprint)
  3. Accepted: April 26, 2022
  4. Accepted Manuscript published: April 27, 2022 (version 1)
  5. Version of Record published: May 11, 2022 (version 2)

Copyright

© 2022, Xiao 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.

Metrics

  • 1,101
    views
  • 285
    downloads
  • 11
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Han Xiao
  2. Tao Zhang
  3. Chang Jun Li
  4. Yong Cao
  5. Lin Feng Wang
  6. Hua Bin Chen
  7. Sheng Can Li
  8. Chang Biao Guan
  9. Jian Zhong Hu
  10. Di Chen
  11. Can Chen
  12. Hong Bin Lu
(2022)
Mechanical stimulation promotes enthesis injury repair by mobilizing Prrx1+ cells via ciliary TGF-β signaling
eLife 11:e73614.
https://doi.org/10.7554/eLife.73614

Share this article

https://doi.org/10.7554/eLife.73614

Categories and tags

Research organism

Further reading

    1. Cell Biology
    2. Stem Cells and Regenerative Medicine

    Differential regulation by CD47 and thrombospondin-1 of extramedullary erythropoiesis in mouse spleen

    Rajdeep Banerjee, Thomas J Meyer ... David D Roberts
    Research Article

    Extramedullary erythropoiesis is not expected in healthy adult mice, but erythropoietic gene expression was elevated in lineage-depleted spleen cells from Cd47−/− mice. Expression of several genes associated with early stages of erythropoiesis was elevated in mice lacking CD47 or its signaling ligand thrombospondin-1, consistent with previous evidence that this signaling pathway inhibits expression of multipotent stem cell transcription factors in spleen. In contrast, cells expressing markers of committed erythroid progenitors were more abundant in Cd47−/− spleens but significantly depleted in Thbs1−/− spleens. Single-cell transcriptome and flow cytometry analyses indicated that loss of CD47 is associated with accumulation and increased proliferation in spleen of Ter119−CD34+ progenitors and Ter119+CD34− committed erythroid progenitors with elevated mRNA expression of Kit, Ermap, and Tfrc. Induction of committed erythroid precursors is consistent with the known function of CD47 to limit the phagocytic removal of aged erythrocytes. Conversely, loss of thrombospondin-1 delays the turnover of aged red blood cells, which may account for the suppression of committed erythroid precursors in Thbs1−/− spleens relative to basal levels in wild-type mice. In addition to defining a role for CD47 to limit extramedullary erythropoiesis, these studies reveal a thrombospondin-1-dependent basal level of extramedullary erythropoiesis in adult mouse spleen.

    1. Developmental Biology
    2. Stem Cells and Regenerative Medicine

    Endothelial cell signature in muscle stem cells validated by VEGFA-FLT1-AKT1 axis promoting survival of muscle stem cell

    Mayank Verma, Yoko Asakura ... Atsushi Asakura
    Research Article Updated

    Endothelial and skeletal muscle lineages arise from common embryonic progenitors. Despite their shared developmental origin, adult endothelial cells (ECs) and muscle stem cells (MuSCs; satellite cells) have been thought to possess distinct gene signatures and signaling pathways. Here, we shift this paradigm by uncovering how adult MuSC behavior is affected by the expression of a subset of EC transcripts. We used several computational analyses including single-cell RNA-seq (scRNA-seq) to show that MuSCs express low levels of canonical EC markers in mice. We demonstrate that MuSC survival is regulated by one such prototypic endothelial signaling pathway (VEGFA-FLT1). Using pharmacological and genetic gain- and loss-of-function studies, we identify the FLT1-AKT1 axis as the key effector underlying VEGFA-mediated regulation of MuSC survival. All together, our data support that the VEGFA-FLT1-AKT1 pathway promotes MuSC survival during muscle regeneration, and highlights how the minor expression of select transcripts is sufficient for affecting cell behavior.

    1. Biochemistry and Chemical Biology
    2. Stem Cells and Regenerative Medicine

    Targeted protein degradation systems to enhance Wnt signaling

    Parthasarathy Sampathkumar, Heekyung Jung ... Yang Li
    Research Article

    Molecules that facilitate targeted protein degradation (TPD) offer great promise as novel therapeutics. The human hepatic lectin asialoglycoprotein receptor (ASGR) is selectively expressed on hepatocytes. We have previously engineered an anti-ASGR1 antibody-mutant RSPO2 (RSPO2RA) fusion protein (called SWEETS) to drive tissue-specific degradation of ZNRF3/RNF43 E3 ubiquitin ligases, which achieved hepatocyte-specific enhanced Wnt signaling, proliferation, and restored liver function in mouse models, and an antibody–RSPO2RA fusion molecule is currently in human clinical trials. In the current study, we identified two new ASGR1- and ASGR1/2-specific antibodies, 8M24 and 8G8. High-resolution crystal structures of ASGR1:8M24 and ASGR2:8G8 complexes revealed that these antibodies bind to distinct epitopes on opposing sides of ASGR, away from the substrate-binding site. Both antibodies enhanced Wnt activity when assembled as SWEETS molecules with RSPO2RA through specific effects sequestering E3 ligases. In addition, 8M24-RSPO2RA and 8G8-RSPO2RA efficiently downregulate ASGR1 through TPD mechanisms. These results demonstrate the possibility of combining different therapeutic effects and degradation mechanisms in a single molecule.