TY - JOUR
T1 - CCN4/WISP-1 positively regulates chondrogenesis by controlling TGF-β3 function
AU - Yoshioka, Yuya
AU - Ono, Mitsuaki
AU - Maeda, Azusa
AU - Kilts, Tina M.
AU - Hara, Emilio Satoshi
AU - Khattab, Hany
AU - Ueda, Junji
AU - Aoyama, Eriko
AU - Oohashi, Toshitaka
AU - Takigawa, Masaharu
AU - Young, Marian F.
AU - Kuboki, Takuo
N1 - Funding Information:
This study was supported in part by JSPS KAKENHI Grant Numbers # 23890122 and # 26861639 and the Intramural Program of the NIH, NIDCR.
Publisher Copyright:
© 2015 Elsevier Inc.
PY - 2016/2/1
Y1 - 2016/2/1
N2 - The CCN family of proteins plays important roles in development and homeostasis of bone and cartilage. To understand the role of CCN4 in chondrogenesis, human bone marrow stromal cells (hBMSCs) were transduced with CCN4 adenovirus (adCCN4) or siRNA to CCN4 (siCCN4) in the presence or absence of transforming growth factor-β3 (TGF-β3). Overexpression of CCN4 enhanced TGF-β3-induced SMAD2/3 phosphorylation and chondrogenesis of hBMSCs in an in vitro assay using a micromass culture model. On the other hand, knockdown of CCN4 inhibited the TGF-β3-induced SMAD2/3 phosphorylation and synthesis of cartilage matrix in micromass cultures of hBMSCs. Immunoprecipitation-western blot analysis revealed that CCN4 bound to TGF-β3 and regulated the ability of TGF-β3 to bind to hBMSCs. In vivo analysis confirmed there was a significant decrease in the gene expression levels of chondrocyte markers in cartilage samples from Ccn4-knock out (KO) mice, compared to those from wild type (WT) control. In order to investigate the regenerative properties of the articular cartilage in Ccn4-KO mice, articular cartilage defects were surgically performed in the knee joints of young mice, and the results showed that the cartilage was partially repaired in WT mice, but not in Ccn4-KO mice. In conclusion, these results show, for the first time, that CCN4 has a positive influence on chondrogenic differentiation by modulating the effects of TGF-β3.
AB - The CCN family of proteins plays important roles in development and homeostasis of bone and cartilage. To understand the role of CCN4 in chondrogenesis, human bone marrow stromal cells (hBMSCs) were transduced with CCN4 adenovirus (adCCN4) or siRNA to CCN4 (siCCN4) in the presence or absence of transforming growth factor-β3 (TGF-β3). Overexpression of CCN4 enhanced TGF-β3-induced SMAD2/3 phosphorylation and chondrogenesis of hBMSCs in an in vitro assay using a micromass culture model. On the other hand, knockdown of CCN4 inhibited the TGF-β3-induced SMAD2/3 phosphorylation and synthesis of cartilage matrix in micromass cultures of hBMSCs. Immunoprecipitation-western blot analysis revealed that CCN4 bound to TGF-β3 and regulated the ability of TGF-β3 to bind to hBMSCs. In vivo analysis confirmed there was a significant decrease in the gene expression levels of chondrocyte markers in cartilage samples from Ccn4-knock out (KO) mice, compared to those from wild type (WT) control. In order to investigate the regenerative properties of the articular cartilage in Ccn4-KO mice, articular cartilage defects were surgically performed in the knee joints of young mice, and the results showed that the cartilage was partially repaired in WT mice, but not in Ccn4-KO mice. In conclusion, these results show, for the first time, that CCN4 has a positive influence on chondrogenic differentiation by modulating the effects of TGF-β3.
KW - Articular cartilage
KW - Bone marrow stromal cell (BMSC)
KW - CCN4/WISP-1
KW - Chondrogenesis
KW - TGF-β3
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U2 - 10.1016/j.bone.2015.11.007
DO - 10.1016/j.bone.2015.11.007
M3 - Article
C2 - 26555637
AN - SCOPUS:84947777296
SN - 8756-3282
VL - 83
SP - 162
EP - 170
JO - Bone
JF - Bone
ER -
