FOXO1 transcription factor regulates chondrogenic differentiation through transforming growth factor β1 signaling

Ichiro Kurakazu, Yukio Akasaki, Mitsumasa Hayashida, Hidetoshi Tsushima, Norio Goto, Takuya Sueishi, Masakazu Toya, Masanari Kuwahara, Ken Okazaki, Tomas Duffy, Martin K. Lotz, Yasuharu Nakashima

Research output: Contribution to journalArticle

Abstract

The forkhead box O (FOXO) proteins are transcription factors involved in the differentiation of many cell types. Type II collagen (Col2) Cre-Foxo1-knockout and Col2-Cre-Foxo1,3,4 triple-knockout mice exhibit growth plate malformation. Moreover, recent studies have reported that in some cells, the expressions and activities of FOXOs are promoted by transforming growth factor β1 (TGFβ1), a growth factor playing a key role in chondrogenic differentiation. Here, using a murine chondrogenic cell line (ATDC5), mouse embryos, and human mesenchymal stem cells, we report the mechanisms by which FOXOs affect chondrogenic differentiation. FOXO1 expression increased along with chondrogenic differentiation, and FOXO1 inhibition suppressed chondrogenic differentiation. TGFβ1/ SMAD signaling promoted expression and activity of FOXO1. In ATDC5, FOXO1 knockdown suppressed expression of sexdetermining region Y box 9 (Sox9), a master regulator of chondrogenic differentiation, resulting in decreased collagen type II α1 (Col2a1) and aggrecan (Acan) expression after TGFβ1 treatment. On the other hand, chemical FOXO1 inhibition suppressed Col2a1 and Acan expression without suppressing Sox9. To investigate the effects of FOXO1 on chondrogenic differentiation independently of SOX9, we examined FOXO1's effects on the cell cycle. FOXO1 inhibition suppressed expression of p21 and cell-cycle arrest in G0/G1 phase. Conversely, FOXO1 overexpression promoted expression of p21 and cell-cycle arrest. FOXO1 inhibition suppressed expression of nascent p21 RNAby TGFβ1, andFOXO1bound the p21 promoter. p21 inhibition suppressed expression of Col2a1 and Acan during chondrogenic differentiation. These results suggest that FOXO1 is necessary for not only SOX9 expression, but also cellcycle arrest during chondrogenic differentiation via TGFβ1 signaling.

Original languageEnglish
Pages (from-to)17555-17569
Number of pages15
JournalJournal of Biological Chemistry
Volume294
Issue number46
DOIs
Publication statusPublished - Jan 1 2019

Fingerprint

Transforming Growth Factors
Transcription Factors
Aggrecans
Cells
Collagen Type II
Cell Cycle Checkpoints
Forkhead Transcription Factors
Cell Cycle Resting Phase
Growth Plate
G1 Phase
Collagen Type I
Stem cells
Mesenchymal Stromal Cells
Knockout Mice
Cell Differentiation
Intercellular Signaling Peptides and Proteins
Cell Cycle
Embryonic Structures
Cell Line
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

FOXO1 transcription factor regulates chondrogenic differentiation through transforming growth factor β1 signaling. / Kurakazu, Ichiro; Akasaki, Yukio; Hayashida, Mitsumasa; Tsushima, Hidetoshi; Goto, Norio; Sueishi, Takuya; Toya, Masakazu; Kuwahara, Masanari; Okazaki, Ken; Duffy, Tomas; Lotz, Martin K.; Nakashima, Yasuharu.

In: Journal of Biological Chemistry, Vol. 294, No. 46, 01.01.2019, p. 17555-17569.

Research output: Contribution to journalArticle

Kurakazu, Ichiro ; Akasaki, Yukio ; Hayashida, Mitsumasa ; Tsushima, Hidetoshi ; Goto, Norio ; Sueishi, Takuya ; Toya, Masakazu ; Kuwahara, Masanari ; Okazaki, Ken ; Duffy, Tomas ; Lotz, Martin K. ; Nakashima, Yasuharu. / FOXO1 transcription factor regulates chondrogenic differentiation through transforming growth factor β1 signaling. In: Journal of Biological Chemistry. 2019 ; Vol. 294, No. 46. pp. 17555-17569.
@article{7cf6945944a547babedd06d21e449a2d,
title = "FOXO1 transcription factor regulates chondrogenic differentiation through transforming growth factor β1 signaling",
abstract = "The forkhead box O (FOXO) proteins are transcription factors involved in the differentiation of many cell types. Type II collagen (Col2) Cre-Foxo1-knockout and Col2-Cre-Foxo1,3,4 triple-knockout mice exhibit growth plate malformation. Moreover, recent studies have reported that in some cells, the expressions and activities of FOXOs are promoted by transforming growth factor β1 (TGFβ1), a growth factor playing a key role in chondrogenic differentiation. Here, using a murine chondrogenic cell line (ATDC5), mouse embryos, and human mesenchymal stem cells, we report the mechanisms by which FOXOs affect chondrogenic differentiation. FOXO1 expression increased along with chondrogenic differentiation, and FOXO1 inhibition suppressed chondrogenic differentiation. TGFβ1/ SMAD signaling promoted expression and activity of FOXO1. In ATDC5, FOXO1 knockdown suppressed expression of sexdetermining region Y box 9 (Sox9), a master regulator of chondrogenic differentiation, resulting in decreased collagen type II α1 (Col2a1) and aggrecan (Acan) expression after TGFβ1 treatment. On the other hand, chemical FOXO1 inhibition suppressed Col2a1 and Acan expression without suppressing Sox9. To investigate the effects of FOXO1 on chondrogenic differentiation independently of SOX9, we examined FOXO1's effects on the cell cycle. FOXO1 inhibition suppressed expression of p21 and cell-cycle arrest in G0/G1 phase. Conversely, FOXO1 overexpression promoted expression of p21 and cell-cycle arrest. FOXO1 inhibition suppressed expression of nascent p21 RNAby TGFβ1, andFOXO1bound the p21 promoter. p21 inhibition suppressed expression of Col2a1 and Acan during chondrogenic differentiation. These results suggest that FOXO1 is necessary for not only SOX9 expression, but also cellcycle arrest during chondrogenic differentiation via TGFβ1 signaling.",
author = "Ichiro Kurakazu and Yukio Akasaki and Mitsumasa Hayashida and Hidetoshi Tsushima and Norio Goto and Takuya Sueishi and Masakazu Toya and Masanari Kuwahara and Ken Okazaki and Tomas Duffy and Lotz, {Martin K.} and Yasuharu Nakashima",
year = "2019",
month = "1",
day = "1",
doi = "10.1074/jbc.RA119.009409",
language = "English",
volume = "294",
pages = "17555--17569",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "46",

}

TY - JOUR

T1 - FOXO1 transcription factor regulates chondrogenic differentiation through transforming growth factor β1 signaling

AU - Kurakazu, Ichiro

AU - Akasaki, Yukio

AU - Hayashida, Mitsumasa

AU - Tsushima, Hidetoshi

AU - Goto, Norio

AU - Sueishi, Takuya

AU - Toya, Masakazu

AU - Kuwahara, Masanari

AU - Okazaki, Ken

AU - Duffy, Tomas

AU - Lotz, Martin K.

AU - Nakashima, Yasuharu

PY - 2019/1/1

Y1 - 2019/1/1

N2 - The forkhead box O (FOXO) proteins are transcription factors involved in the differentiation of many cell types. Type II collagen (Col2) Cre-Foxo1-knockout and Col2-Cre-Foxo1,3,4 triple-knockout mice exhibit growth plate malformation. Moreover, recent studies have reported that in some cells, the expressions and activities of FOXOs are promoted by transforming growth factor β1 (TGFβ1), a growth factor playing a key role in chondrogenic differentiation. Here, using a murine chondrogenic cell line (ATDC5), mouse embryos, and human mesenchymal stem cells, we report the mechanisms by which FOXOs affect chondrogenic differentiation. FOXO1 expression increased along with chondrogenic differentiation, and FOXO1 inhibition suppressed chondrogenic differentiation. TGFβ1/ SMAD signaling promoted expression and activity of FOXO1. In ATDC5, FOXO1 knockdown suppressed expression of sexdetermining region Y box 9 (Sox9), a master regulator of chondrogenic differentiation, resulting in decreased collagen type II α1 (Col2a1) and aggrecan (Acan) expression after TGFβ1 treatment. On the other hand, chemical FOXO1 inhibition suppressed Col2a1 and Acan expression without suppressing Sox9. To investigate the effects of FOXO1 on chondrogenic differentiation independently of SOX9, we examined FOXO1's effects on the cell cycle. FOXO1 inhibition suppressed expression of p21 and cell-cycle arrest in G0/G1 phase. Conversely, FOXO1 overexpression promoted expression of p21 and cell-cycle arrest. FOXO1 inhibition suppressed expression of nascent p21 RNAby TGFβ1, andFOXO1bound the p21 promoter. p21 inhibition suppressed expression of Col2a1 and Acan during chondrogenic differentiation. These results suggest that FOXO1 is necessary for not only SOX9 expression, but also cellcycle arrest during chondrogenic differentiation via TGFβ1 signaling.

AB - The forkhead box O (FOXO) proteins are transcription factors involved in the differentiation of many cell types. Type II collagen (Col2) Cre-Foxo1-knockout and Col2-Cre-Foxo1,3,4 triple-knockout mice exhibit growth plate malformation. Moreover, recent studies have reported that in some cells, the expressions and activities of FOXOs are promoted by transforming growth factor β1 (TGFβ1), a growth factor playing a key role in chondrogenic differentiation. Here, using a murine chondrogenic cell line (ATDC5), mouse embryos, and human mesenchymal stem cells, we report the mechanisms by which FOXOs affect chondrogenic differentiation. FOXO1 expression increased along with chondrogenic differentiation, and FOXO1 inhibition suppressed chondrogenic differentiation. TGFβ1/ SMAD signaling promoted expression and activity of FOXO1. In ATDC5, FOXO1 knockdown suppressed expression of sexdetermining region Y box 9 (Sox9), a master regulator of chondrogenic differentiation, resulting in decreased collagen type II α1 (Col2a1) and aggrecan (Acan) expression after TGFβ1 treatment. On the other hand, chemical FOXO1 inhibition suppressed Col2a1 and Acan expression without suppressing Sox9. To investigate the effects of FOXO1 on chondrogenic differentiation independently of SOX9, we examined FOXO1's effects on the cell cycle. FOXO1 inhibition suppressed expression of p21 and cell-cycle arrest in G0/G1 phase. Conversely, FOXO1 overexpression promoted expression of p21 and cell-cycle arrest. FOXO1 inhibition suppressed expression of nascent p21 RNAby TGFβ1, andFOXO1bound the p21 promoter. p21 inhibition suppressed expression of Col2a1 and Acan during chondrogenic differentiation. These results suggest that FOXO1 is necessary for not only SOX9 expression, but also cellcycle arrest during chondrogenic differentiation via TGFβ1 signaling.

UR - http://www.scopus.com/inward/record.url?scp=85075089909&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85075089909&partnerID=8YFLogxK

U2 - 10.1074/jbc.RA119.009409

DO - 10.1074/jbc.RA119.009409

M3 - Article

C2 - 31601652

AN - SCOPUS:85075089909

VL - 294

SP - 17555

EP - 17569

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 46

ER -