Mesenchymal-to-epithelial transition of osteoblasts induced by Fam20c knockout

Geng, Yawei; Zhang, Zhen; Jin, Han; Da, Junlong; Zhang, Kai; Wang, Jianqun; Guo, Yu-Yao; Zhang, Bin; Liu, Ying

doi:10.1007/s13258-021-01170-4

Cited by 3 publications

(8 citation statements)

References 40 publications

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“…These findings align with previous findings showing that Fam20c deficiency in mouse dental mesenchymal cells resulted in suppressed proliferation, mobility, and wound healing (Liu, Zhou, et al., 2018). Similarly, FAM20C knockout in osteoblast cell lines presented reduced cell proliferation, adhesion, migration, and colony (Geng et al., 2022; Liu, Zhang, et al., 2018; Tagliabracci et al., 2015). These data strongly suggest that FAM20A plays a crucial role in regulating the behaviours of dental mesenchymal cells that are essential for their appropriate function and tissue development.…”

Section: Discussionmentioning

confidence: 98%

In‐depth investigation of FAM20A insufficiency effects on deciduous dental pulp cells: Altered behaviours, osteogenic differentiation, and inflammatory gene expression

Sriwattanapong,

Theerapanon,

Khamwachirapitak

et al. 2024

Int Endodontic J

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AimLoss‐of‐function mutations in FAM20A result in amelogenesis imperfecta IG (AI1G) or enamel‐renal syndrome, characterized by hypoplastic enamel, ectopic calcification, and gingival hyperplasia, with some cases reporting spontaneous tooth infection. Despite previous reports on the consequence of FAM20A reduction in gingival fibroblasts and transcriptome analyses of AI1G pulp tissues, suggesting its involvement in mineralization and infection, its role in deciduous dental pulp cells (DDP) remains unreported. The aim of this study was to evaluate the properties of DDP obtained from an AI1G patient, providing additional insights into the effects of FAM20A on the mineralization of DDP.MethodologyDDP were obtained from a FAM20A‐AI1G patient (mutant cells) and three healthy individuals. Cellular behaviours were examined using flow cytometry, MTT, attachment and spreading, colony formation, and wound healing assays. Osteogenic induction was applied to DDP, followed by alizarin red S staining to assess their osteogenic differentiation. The expression of FAM20A‐related genes, osteogenic genes, and inflammatory genes was analysed using real‐time PCR, Western blot, and/or immunolocalization. Additionally, STRING analysis was performed to predict potential protein–protein interaction networks.ResultsThe mutant cells exhibited a significant reduction in FAM20A mRNA and protein levels, as well as proliferation, migration, attachment, and colony formation. However, normal FAM20A subcellular localization was maintained. Additionally, osteogenic/odontogenic genes, OSX, OPN, RUNX2, BSP, and DSPP, were downregulated, along with upregulated ALP. STRING analysis suggested a potential correlation between FAM20A and these osteogenic genes. After osteogenic induction, the mutant cells demonstrated reduced mineral deposition and dysregulated expression of osteogenic genes. Remarkably, FAM20A, FAM20C, RUNX2, OPN, and OSX were significantly upregulated in the mutant cells, whilst ALP, and OCN was downregulated. Furthermore, the mutant cells exhibited a significant increase in inflammatory gene expression, that is, IL‐1β and TGF‐β1, whereas IL‐6 and NFκB1 expression was significantly reduced.ConclusionThe reduction of FAM20A in mutant DDP is associated with various cellular deficiencies, including delayed proliferation, attachment, spreading, and migration as well as altered osteogenic and inflammatory responses. These findings provide novel insights into the biology of FAM20A in dental pulp cells and shed light on the molecular mechanisms underlying AI1G pathology.

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Section: Discussionmentioning

confidence: 98%

In‐depth investigation of FAM20A insufficiency effects on deciduous dental pulp cells: Altered behaviours, osteogenic differentiation, and inflammatory gene expression

Sriwattanapong,

Theerapanon,

Khamwachirapitak

et al. 2024

Int Endodontic J

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“…Immortalized mouse Fam20c f/f osteoblast cells (referred as “OB Fam20c f/f ”) were purchased from Applied Biological Materials Inc. (Lot# T0038). Fam20c -deficient osteoblasts (referred as “OB Fam20c KO ”) were established with infection of Cre recombinase lentivirus as previously published work described [ 28 ]. OB Fam20c f/f and OB Fam20c KO were cultured at 37 °C with 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

“…RNA-seq analysis was as previously reported [ 28 ]. The associated genes in open chromatin regions with enhanced and attenuated ATAC-seq signal overlapped with the up and down differentially expression genes (DEGs) in the transcriptomes, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Zhang et al [ 23 ] analyzed the proteins that interact with FAM20C in the endoplasmic reticulum (ER) and Golgi apparatus through immunoprecipitation and mass spectrometry, revealing that FAM20C could phosphorylate ER oxidoreductin 1α (Ero1α) to maintain the ER redox homeostasis. Besides, our previous research conducted a transcriptional analysis of Fam20c -deficient osteoblasts, indicating the cells undergo mesenchymal to epithelial transformation [ 24 ]. Besides a single sequencing method, the combination of multi-omics analysis provides a more comprehensive picture linking “molecular information” to “phenotype” through functional and signaling networks [ 25 – 27 ].…”

Section: Introductionmentioning

confidence: 99%

“…Here, we use established Fam20c -deficient cells in our previously published work [ 28 ] to conduct a comprehensive omics analysis. To probe the relationship between chromatin, transcriptional and protein changes response to Fam20c ablation in osteoblasts, we performed integrated omics analyses using an assay for transposase-accessible chromatin using sequencing (ATAC-seq), RNA sequencing (RNA-seq), proteomics, and phosphoproteomics.…”

Section: Introductionmentioning

confidence: 99%

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Fam20c regulates the calpain proteolysis system through phosphorylating Calpasatatin to maintain cell homeostasis

Liu

Zhang

et al. 2023

J Transl Med

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Background The family with sequence similarity 20-member C (FAM20C) kinase, a Golgi casein kinase, which is responsible for phosphorylating the majority of the extracellular phosphoproteins within S-x-E/pS motifs, and is fundamentally associated with multiple biological processes to maintain cell proliferation, biomineralization, migration, adhesion, and phosphate homeostasis. In dissecting how FAM20C regulates downstream molecules and potential mechanisms, however, there are multiple target molecules of FAM20C, particularly many phenomena remain elusive, such as changes in cell-autonomous behaviors, incompatibility in genotypes and phenotypes, and others. Methods Here, assay for transposase-accessible chromatin using sequencing (ATAC-seq), RNA sequencing (RNA-seq), proteomics, and phosphoproteomics were performed in Fam20c-dificient osteoblasts and to facilitate an integrated analysis and determine the impact of chromatin accessibility, genomic expression, protein alterations, signaling pathway, and post translational modifcations. Results By combining ATAC-seq and RNA-seq, we identified TCF4 and Wnt signaling pathway as the key regulators in Fam20c-dificient cells. Further, we showed Calpastatin/Calpain proteolysis system as a novel target axis for FAM20C to regulate cell migration and F-actin cytoskeleton by integrated analysis of proteomics and phosphoproteomics. Furthermore, Calpastatin/Calpain proteolysis system could negatively regulate the Wnt signaling pathway. Conclusion These observations implied that Fam20c knockout osteoblasts would cause cell homeostatic imbalance, involving changes in multiple signaling pathways in the conduction system.

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Knocking out FAM20C in pre-osteoblasts leads to up-regulation of osteoclast differentiation to affect long bone development

Jiang,

Liu,

Liu

et al. 2024

Gene

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Mesenchymal-to-epithelial transition of osteoblasts induced by Fam20c knockout

Cited by 3 publications

References 40 publications

In‐depth investigation of FAM20A insufficiency effects on deciduous dental pulp cells: Altered behaviours, osteogenic differentiation, and inflammatory gene expression

In‐depth investigation of FAM20A insufficiency effects on deciduous dental pulp cells: Altered behaviours, osteogenic differentiation, and inflammatory gene expression

Fam20c regulates the calpain proteolysis system through phosphorylating Calpasatatin to maintain cell homeostasis

Knocking out FAM20C in pre-osteoblasts leads to up-regulation of osteoclast differentiation to affect long bone development

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