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Connective tissue growth factor contributes to joint homeostasis and osteoarthritis severity by controlling the matrix sequestration and activation of latent TGFβ

Tang, Xiaodi ; Muhammad, Hayat ; McLean, Celia ; Miotla-Zarebska, Jadwiga ; Fleming, Jacob ; Didangelos, Athanasios ; Önnerfjord, Patrik LU orcid ; Leask, Andrew ; Saklatvala, Jeremy and Vincent, Tonia L. (2018) In Annals of the Rheumatic Diseases 77(9). p.1372-1380
Abstract

Objectives: One mechanism by which cartilage responds to mechanical load is by releasing heparin-bound growth factors from the pericellular matrix (PCM). By proteomic analysis of the PCM, we identified connective tissue growth factor (CTGF) and here investigate its function and mechanism of action. Methods: Recombinant CTGF (rCTGF) was used to stimulate human chondrocytes for microarray analysis. Endogenous CTGF was investigated by in vitro binding assays and confocal microscopy. Its release from cut cartilage (injury CM) was analysed by Western blot under reducing and non-reducing conditions. A postnatal, conditional CtgfcKO mouse was generated for cartilage injury experiments and to explore the course of osteoarthritis (OA)... (More)

Objectives: One mechanism by which cartilage responds to mechanical load is by releasing heparin-bound growth factors from the pericellular matrix (PCM). By proteomic analysis of the PCM, we identified connective tissue growth factor (CTGF) and here investigate its function and mechanism of action. Methods: Recombinant CTGF (rCTGF) was used to stimulate human chondrocytes for microarray analysis. Endogenous CTGF was investigated by in vitro binding assays and confocal microscopy. Its release from cut cartilage (injury CM) was analysed by Western blot under reducing and non-reducing conditions. A postnatal, conditional CtgfcKO mouse was generated for cartilage injury experiments and to explore the course of osteoarthritis (OA) by destabilisation of the medial meniscus. siRNA knockdown was performed on isolated human chondrocytes. Results: The biological responses of rCTGF were TGFβ dependent. CTGF displaced latent TGFβ from cartilage and both were released on cartilage injury. CTGF and latent TGFβ migrated as a single high molecular weight band under non-reducing conditions, suggesting that they were in a covalent (disulfide) complex. This was confirmed by immunoprecipitation. Using CtgfcKO mice, CTGF was required for sequestration of latent TGFβ in the matrix and activation of the latent complex at the cell surface through TGFβR3. In vivo deletion of CTGF increased the thickness of the articular cartilage and protected mice from OA. Conclusions: CTGF is a latent TGFβ binding protein that controls the matrix sequestration and activation of TGFβ in cartilage. Deletion of CTGF in vivo caused a paradoxical increase in Smad2 phosphorylation resulting in thicker cartilage that was protected from OA.

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author
; ; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
arthritis, chondrocytes, osteoarthritis
in
Annals of the Rheumatic Diseases
volume
77
issue
9
pages
1372 - 1380
publisher
BMJ Publishing Group
external identifiers
  • scopus:85048837141
  • pmid:29925506
ISSN
0003-4967
DOI
10.1136/annrheumdis-2018-212964
language
English
LU publication?
yes
id
58e7259f-95ab-42a2-8564-cca86518d4dc
date added to LUP
2018-07-05 12:49:37
date last changed
2024-04-01 07:57:31
@article{58e7259f-95ab-42a2-8564-cca86518d4dc,
  abstract     = {{<p>Objectives: One mechanism by which cartilage responds to mechanical load is by releasing heparin-bound growth factors from the pericellular matrix (PCM). By proteomic analysis of the PCM, we identified connective tissue growth factor (CTGF) and here investigate its function and mechanism of action. Methods: Recombinant CTGF (rCTGF) was used to stimulate human chondrocytes for microarray analysis. Endogenous CTGF was investigated by in vitro binding assays and confocal microscopy. Its release from cut cartilage (injury CM) was analysed by Western blot under reducing and non-reducing conditions. A postnatal, conditional Ctgf<sup>cKO</sup> mouse was generated for cartilage injury experiments and to explore the course of osteoarthritis (OA) by destabilisation of the medial meniscus. siRNA knockdown was performed on isolated human chondrocytes. Results: The biological responses of rCTGF were TGFβ dependent. CTGF displaced latent TGFβ from cartilage and both were released on cartilage injury. CTGF and latent TGFβ migrated as a single high molecular weight band under non-reducing conditions, suggesting that they were in a covalent (disulfide) complex. This was confirmed by immunoprecipitation. Using Ctgf<sup>cKO</sup> mice, CTGF was required for sequestration of latent TGFβ in the matrix and activation of the latent complex at the cell surface through TGFβR3. In vivo deletion of CTGF increased the thickness of the articular cartilage and protected mice from OA. Conclusions: CTGF is a latent TGFβ binding protein that controls the matrix sequestration and activation of TGFβ in cartilage. Deletion of CTGF in vivo caused a paradoxical increase in Smad2 phosphorylation resulting in thicker cartilage that was protected from OA.</p>}},
  author       = {{Tang, Xiaodi and Muhammad, Hayat and McLean, Celia and Miotla-Zarebska, Jadwiga and Fleming, Jacob and Didangelos, Athanasios and Önnerfjord, Patrik and Leask, Andrew and Saklatvala, Jeremy and Vincent, Tonia L.}},
  issn         = {{0003-4967}},
  keywords     = {{arthritis; chondrocytes; osteoarthritis}},
  language     = {{eng}},
  month        = {{06}},
  number       = {{9}},
  pages        = {{1372--1380}},
  publisher    = {{BMJ Publishing Group}},
  series       = {{Annals of the Rheumatic Diseases}},
  title        = {{Connective tissue growth factor contributes to joint homeostasis and osteoarthritis severity by controlling the matrix sequestration and activation of latent TGFβ}},
  url          = {{http://dx.doi.org/10.1136/annrheumdis-2018-212964}},
  doi          = {{10.1136/annrheumdis-2018-212964}},
  volume       = {{77}},
  year         = {{2018}},
}