Summary
Objectives
ANP32A is a key protector of cartilage health, via preventing oxidative stress and
Wnt hyper-activation. We aimed to unravel how ANP32A is regulated in cartilage.
Methods
A bioinformatics pipeline was applied to identify regulators of ANP32A. Pathways of interest were targeted to study their impact on ANP32A in in vitro cultures of the human chondrocyte C28/I2 cell-line and primary human articular chondrocytes
(hACs) from up to five different donors, using Wnt-activator CHIR99021, hypoxia-mimetic
IOX2 and a hypoxia chamber. ANP32A was evaluated using real-time quantitative polymerase
chain reaction (RT-qPCR) and Western blot. In vivo, the effect of hypoxia was examined by immunohistochemistry in mice injected intra-articularly
with IOX2 after destabilization of the medial meniscus. Effects of Wnt hyper-activation
were investigated using Frzb-knockout mice and wild-type mice treated intra-articularly with CHIR99021. Wnt inhibition
effects were assessed upon intra-articular injection of XAV939.
Results
The hypoxia and Wnt signaling pathways were identified as networks controlling ANP32A
expression. In vitro and in vivo experiments demonstrated increases in ANP32A upon hypoxic conditions (1.3-fold in
hypoxia in C28/I2 cells with 95% confidence interval (CI) [1.11–1.54] and 1.90-fold
in hACs [95% CI: 1.56–2] and 1.67-fold in ANP32A protein levels after DMM surgery
with IOX2 injections [95% CI: 1.33–2.08]). Wnt hyper-activation decreased ANP32A in
chondrocytes in vitro (1.23-fold decrease [95% CI: 1.02–1.49]) and in mice (1.45-fold decrease after CHIR99021
injection [95% CI: 1.22–1.72] and 1.41-fold decrease in Frzb-knockout mice [95% CI: 1.00–1.96]). Hypoxia and Wnt modulated ataxia-telangiectasia
mutated serine/threonine kinase (ATM), an ANP32A target gene, in hACs (1.89-fold increase [95% CI: 1.38–2.60] and 1.41-fold
decrease [95% CI: 1.02–1.96]).
Conclusions
Maintaining hypoxia and limiting Wnt activation sustain ANP32A and protect against osteoarthritis.
Keywords
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Article info
Publication history
Published online: November 09, 2022
Accepted:
October 21,
2022
Received:
June 30,
2022
Identification
Copyright
© 2022 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.
