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SERPINA3 is a marker of cartilage differentiation and is essential for the expression of extracellular matrix genes during early chondrogenesis

Lookup NU author(s): Dr Matthew Barter, Dr Sarah RiceORCiD, Hua Lin, Dr Adrian Falconer, Dr Jamie Soul, Dr Maria Arques Mengual, Emeritus Professor Nick Europe-Finner, Emeritus Professor Drew Rowan, Professor David YoungORCiD, Dr David Wilkinson

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

© 2024Serine proteinase inhibitors (serpins) are a family of structurally similar proteins which regulate many diverse biological processes from blood coagulation to extracellular matrix (ECM) remodelling. Chondrogenesis involves the condensation and differentiation of mesenchymal stem cells (MSCs) into chondrocytes which occurs during early development. Here, and for the first time, we demonstrate that one serpin, SERPINA3 (gene name SERPINA3, protein also known as alpha-1 antichymotrypsin), plays a critical role in chondrogenic differentiation. We observed that SERPINA3 expression was markedly induced at early time points during in vitro chondrogenesis. We examined the expression of SERPINA3 in human cartilage development, identifying significant enrichment of SERPINA3 in developing cartilage compared to total limb, which correlated with well-described markers of cartilage differentiation. When SERPINA3 was silenced using siRNA, cartilage pellets were smaller and contained lower proteoglycan as determined by dimethyl methylene blue assay (DMMB) and safranin-O staining. Consistent with this, RNA sequencing revealed significant downregulation of genes associated with cartilage ECM formation perturbing chondrogenesis. Conversely, SERPINA3 silencing had a negligible effect on the gene expression profile during osteogenesis suggesting the role of SERPINA3 is specific to chondrocyte differentiation. The global effect on cartilage formation led us to investigate the effect of SERPINA3 silencing on the master transcriptional regulator of chondrogenesis, SOX9. Indeed, we observed that SOX9 protein levels were markedly reduced at early time points suggesting a role for SERPINA3 in regulating SOX9 expression and activity. In summary, our data support a non-redundant role for SERPINA3 in enabling chondrogenesis via regulation of SOX9 levels.


Publication metadata

Author(s): Barter MJ, Turner DA, Rice SJ, Hines M, Lin H, Falconer AMD, McDonnell E, Soul J, Arques MDC, Europe-Finner GN, Rowan AD, Young DA, Wilkinson DJ

Publication type: Article

Publication status: Published

Journal: Matrix Biology

Year: 2024

Volume: 133

Pages: 33-42

Print publication date: 01/11/2024

Online publication date: 02/08/2024

Acceptance date: 30/07/2024

Date deposited: 27/08/2024

ISSN (print): 0945-053X

ISSN (electronic): 1569-1802

Publisher: Elsevier B.V.

URL: https://doi.org/10.1016/j.matbio.2024.07.004

DOI: 10.1016/j.matbio.2024.07.004

Data Access Statement: RNA-seq data are available from GEO accession (GSE247491 and GSE247528). Code for bioinformatic analysis is available in the link provided in Experimental Procedures.

PubMed id: 39097037


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Funding

Funder referenceFunder name
22418
22615
Dunhill Medical Trust
MR/P020941/1
JGW Patterson Foundation
JXR10641
MRC-Arthritis Research UK Centre for Integrated Research into Musculoskeletal Ageing (CIMA)
R476/0516Dunhill Medical Trust
RGS\R1\221186
Royal Society
Versus Arthritis

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