FOXC1 negatively regulates BMP-SMAD activity and Id1 expression during osteoblast differentiation

Caddy, Jordan. C.; Luoma, Leiah M.; Berry, Fred B.

doi:10.1101/673889

Cited by 4 publications

(6 citation statements)

References 53 publications

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“…However, there are some opinions against the positive effect of FOXC1 in osteogenesis. Jordan C. Caddy et al described that FOXC1 downregulates BMP4-induced osteoblast differentiation 34 . FOXC1 expression is regulated by microRNAs, such as miR-138-5p, miR-374c and miR-133 35,36 .…”

Section: Discussionmentioning

confidence: 99%

The miR-204-5p/FOXC1/GDF7 axis regulates the osteogenic differentiation of human adipose-derived stem cells via the AKT and p38 signalling pathways

Zhou

Liu

Wang

et al. 2020

Preprint

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Background: Human adipose-derived stem cells (hADSCs) are stem cells with the potential to differentiate in multiple directions. MiR-204-5p is expressed at low levels during the osteogenic differentiation of hADSCs, and its specific regulatory mechanism remains unclear. Here, we aimed to explore the function and possible molecular mechanism of miR-204-5p in the osteogenic differentiation of hADSCs.Methods: The expression patterns of miR-204-5p, Runx2, alkaline phosphatase (ALP), osteocalcin (OCN), forkhead box C1 (FOXC1) and growth differentiation factor 7 (GDF7) in hADSCs during osteogenesis were detected by qRT-PCR. Then, ALP and alizarin red staining (ARS) were used to detect osteoblast activitie and mineral deposition. Western blotting was conducted to confirm the protein levels. The regulatory relationship among miR-204-5p, FOXC1 and GDF7 was verified by dual-luciferase activity and chromatin immunoprecipitation (ChIP) assays.Results: MiR-204-5p expression was downregulated in hADSC osteogenesis, and overexpression of miR-204-5p suppressed osteogenic differentiation. Furthermore, the levels of FOXC1 and GDF7 were decreased in the miR-204-5p mimics group, which indicates that miR-204-5p overexpression suppresses the expression of FOXC1 and GDF7 by binding to their 3'-untranslated regions (UTRs). Overexpression of FOXC1 or GDF7 improved the inhibition of osteogenic differentiation of hADSCs induced by the miR-204-5p mimics. Moreover, FOXC1 was found to bind to the promoter of miR-204-5p and GDF7, promote the deacetylation of miR-204-5p and reduce the expression of miR-204-5p, thus promoting the expression of GDF7 during osteogenic differentiation. GDF7 induced hADSC osteogenesis differentiation by activating the AKT and P38 signalling pathways.Conclusions: Our results demonstrated that the miR-204-5p/FOXC1/GDF7 axis regulates the osteogenic differentiation of hADSCs via the AKT and p38 signalling pathways. This study further revealed the regulatory mechanism of hADSC differentiation from the perspective of miRNA regulation.

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Section: Discussionmentioning

confidence: 99%

The miR-204-5p/FOXC1/GDF7 axis regulates the osteogenic differentiation of human adipose-derived stem cells via the AKT and p38 signalling pathways

Zhou

Liu

Wang

et al. 2020

Preprint

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“…However, there are some opinions against the positive effect of FOXC1 in osteogenesis. Jordan C. Caddy et al described that FOXC1 would down-regulate suppresses BMP4-induced osteoblast differentiation 34 . And it has been proved that FOXC1 expression is regulated by microRNA, such as miR-138-5p, miR-374c and miR-133 35,36 .…”

Section: Discussionmentioning

confidence: 99%

MiR-204-5p/FOXC1/GDF7 Axis Regulates Osteogenic Differentiation of Human Adipose-Derived Stem Cells Via the AKT and p38 Signaling Pathways

Zhou

Liu

Wang

et al. 2020

Preprint

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Background: Human adipose-derived stem cells (hADSCs) are stem cells with the potential to differentiate in multiple directions. MiR-204-5p is poorly expressed during osteogenic differentiation of hADSCs, and its specific regulatory mechanism remains unclear. Here, we aimed to explore the function and possible molecular mechanism of miR-204-5p involved in the osteogenic differentiation of hADSCs. Methods: The expression pattern of miR-204-5p, Runx2, Alkaline phosphatase (ALP), Osteocalcin (OCN), and Forkhead box C1(FOXC1) and growth differentiation factor 7(GDF7) in hADSCs during osteogenesis were detected by qRT-PCR. Then, ALP and alizarin red staining (ARS) were used to detect the activity of osteoblasts and mineral deposition. Western blot was conducted to confirm the protein levels. The regulation relationship among miR-204-5p, FOXC1 and GDF7 was verified by double luciferase activity and CHIP experiment.Results: First, miR-204-5p expression was down-regulated and overexpressed miR-204-5p suppressed the osteogenic differentiation. Furthermore, the levels of FOXC1 and GDF7 were decreased in the miR-204-5p mimics group, which indicate that overexpressed miR-204-5p would suppress the expression of FOXC1 and GDF7 through binding the 3’UTR region each. Overexpression of FOXC1 or GDF7 could improve the inhibition of osteogenic differentiation of hADSCs induced by the miR-204-5p mimics. Moreover, FOXC1 could bind to the promoter of miR-204-5p and GDF7, promote the deacetylation of miR-204-5p and reduced the expression of miR-204-5p, thus promoting the expression of GDF7 during osteogenic differentiation. GDF7 could induce hADSCs osteogenesis differentiation by activating the AKT and P38 signaling pathways. Conclusions: Our results demonstrated that miR-204-5p/FOXC1/GDF7 axis regulates osteogenic differentiation of hADSCs via the AKT and p38 signaling pathways. This study further understood the regulatory mechanism of hADSCs differentiation balance from the perspective of miRNAs regulation.

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“…RNA (500 ng) was then reverse transcribed to cDNAs using the QuantiTect reverse transcription kit (Qiagen). qRT-PCR analysis was performed as described previously (39) (40). All qRT-PCR experiments were J o u r n a l P r e -p r o o f performed with at least three biological replicates and each contained three technical replicates.…”

Section: Rna-isolation and Qrt-pcrmentioning

confidence: 99%

Loss of Foxc1 and Foxc2 function in chondroprogenitor cells disrupts endochondral ossification

Almubarak

Lavy

Srnic

et al. 2021

Journal of Biological Chemistry

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Endochondral ossification initiates the growth of the majority of the mammalian skeleton and is tightly controlled through gene regulatory networks. The forkhead box transcription factors Foxc1 and Foxc2 regulate aspects of osteoblast function in the formation of the skeleton, but their roles in chondrocytes to control endochondral ossification are less clear. Here, we demonstrate that Foxc1 expression is directly regulated by the activity of SRY (sex-determining region Y)-box 9, one of the earliest transcription factors to specify the chondrocyte lineage. Moreover, we demonstrate that elevated expression of Foxc1 promotes chondrocyte differentiation in mouse embryonic stem cells and loss of Foxc1 function inhibits chondrogenesis in vitro . Using chondrocyte-targeted deletion of Foxc1 and Foxc2 in mice, we reveal a role for these factors in chondrocyte differentiation in vivo . Loss of both Foxc1 and Foxc2 caused a general skeletal dysplasia predominantly affecting the vertebral column. The long bones of the limbs were smaller, mineralization was reduced, and organization of the growth plate was disrupted; in particular, the stacked columnar organization of the proliferative chondrocyte layer was reduced in size and cell proliferation was decreased. Differential gene expression analysis indicated disrupted expression patterns of chondrogenesis and ossification genes throughout the entire process of endochondral ossification in chondrocyte-specific Foxc1/Foxc2 KO embryos. Our results suggest that Foxc1 and Foxc2 are required for normal chondrocyte differentiation and function, as loss of both genes results in disorganization of the growth plate, reduced chondrocyte proliferation, and delays in chondrocyte hypertrophy that prevents ossification of the skeleton.

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FOXC1 negatively regulates BMP-SMAD activity and Id1 expression during osteoblast differentiation

Cited by 4 publications

References 53 publications

The miR-204-5p/FOXC1/GDF7 axis regulates the osteogenic differentiation of human adipose-derived stem cells via the AKT and p38 signalling pathways

The miR-204-5p/FOXC1/GDF7 axis regulates the osteogenic differentiation of human adipose-derived stem cells via the AKT and p38 signalling pathways

MiR-204-5p/FOXC1/GDF7 Axis Regulates Osteogenic Differentiation of Human Adipose-Derived Stem Cells Via the AKT and p38 Signaling Pathways

Loss of Foxc1 and Foxc2 function in chondroprogenitor cells disrupts endochondral ossification

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