Bruton tyrosine kinase (Btk) suppresses osteoblastic differentiation

Kaneshiro, Shoichi; Ebina, Kosuke; Shi, Kenrin; Yoshida, Kiyoshi; Otsuki, Dai; Yoshikawa, Hideki; Higuchi, Chikahisa

doi:10.1007/s00774-014-0612-8

Cited by 4 publications

(4 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…LFM is a potent and selective inhibitor of Bruton’s tyrosine kinase (Btk) [ 51 , 52 , 53 ]. Btk suppresses the osteogenic differentiation of MC3T3-E1 cells, primary calvarial osteoblasts, and bone marrow stromal ST2 cells [ 54 ], which supports the present results. Btk regulates osteoblastic differentiation through the MAPK, NF-κB, and protein kinase C (PKC) α signaling pathways [ 54 ].…”

Section: Discussionsupporting

confidence: 90%

“…Btk suppresses the osteogenic differentiation of MC3T3-E1 cells, primary calvarial osteoblasts, and bone marrow stromal ST2 cells [ 54 ], which supports the present results. Btk regulates osteoblastic differentiation through the MAPK, NF-κB, and protein kinase C (PKC) α signaling pathways [ 54 ]. In addition, Btk is a negative regulator of Wnt–β-catenin signaling in B cells [ 55 ].…”

Section: Discussionsupporting

confidence: 90%

See 1 more Smart Citation

Cell-Based Double-Screening Method to Identify a Reliable Candidate for Osteogenesis-Targeting Compounds

et al. 2022

View full text Add to dashboard Cite

Small-molecule compounds strongly affecting osteogenesis can form the basis of effective therapeutic strategies in bone regenerative medicine. A cell-based high-throughput screening system might be a powerful tool for identifying osteoblast-targeting candidates; however, this approach is generally limited with using only one molecule as a cell-based sensor that does not always reflect the activation of the osteogenic phenotype. In the present study, we used the MC3T3-E1 cell line stably transfected with the green fluorescent protein (GFP) reporter gene driven by a fragment of type I collagen promoter (Col-1a1GFP-MC3T3-E1) to evaluate a double-screening system to identify osteogenic inducible compounds using a combination of a cell-based reporter assay and detection of alkaline phosphatase (ALP) activity. Col-1a1GFP-MC3T3-E1 cells were cultured in an osteogenic induction medium after library screening of 1280 pharmacologically active compounds (Lopack1280). After 7 days, GFP fluorescence was measured using a microplate reader. After 14 days of osteogenic induction, the cells were stained with ALP. Library screening using the Col-1a1/GFP reporter and ALP staining assay detected three candidates with significant osteogenic induction ability. Furthermore, leflunomide, one of the three detected candidates, significantly promoted new bone formation in vivo. Therefore, this double-screening method could identify candidates for osteogenesis-targeting compounds more reliably than conventional methods.

show abstract

Section: Discussionsupporting

confidence: 90%

Section: Discussionsupporting

confidence: 90%

Cell-Based Double-Screening Method to Identify a Reliable Candidate for Osteogenesis-Targeting Compounds

et al. 2022

View full text Add to dashboard Cite

show abstract

“…Integration of methylation and gene expression data corresponding to the Homeobox and bone formation-related genes revealed that both DNA hyper-and hypo-methylation events were associated with both gene downregulation and upregulation ( Figure 2C). Specifically, hypermethylated genes that showed a reduced expression in patient MSCs include positive regulators of OB differentiation such as RUNX2 or NRP2 34 ( Figure 2C) In contrast, negative regulators of osteogenesis such as SFRP2 35 or NFATC2 36 were hypomethylated and consequently upregulated in patient MSCs. In all, these factors could potentially contribute to impaired osteoblastogenesis associated to bone disease in MM and this is summarized in Supplementary Table S7).…”

Section: Aberrant Dna Methylation Is Associated With Differential Hommentioning

confidence: 99%

Targeting aberrant DNA methylation in mesenchymal stromal cells as a treatment for myeloma bone disease

García-Gómez

Rodríguez-Ubreva

et al. 2019

Preprint

View full text Add to dashboard Cite

and agarciag@carrerasresearch.org Running title: Reverting DNA methylation to treat MM bone disease Abbreviations: mesenchymal stromal cells, osteoblast, multiple myeloma, DNA methylation Abstract word count: 212 Word count: 3999 Figure count: 5 Reference count: 60 ABSTRACT Multiple myeloma (MM) progression and myeloma-associated bone disease (MBD) are highly dependent on the bone marrow (BM) microenvironment, in particular on mesenchymal stromal cells (MSCs). MSCs from MM patients exhibit an abnormal transcriptional profile, suggesting that epigenetic alterations could be governing the tumor-promoting functions of MSCs and their prolonged osteoblast (OB) suppression in MM. In this study, we analyzed the DNA methylome of BM-derived MSCs from patients with monoclonal gammopathy of undetermined significance, smoldering myeloma and symptomatic MM at diagnosis in comparison with their normal counterparts. DNA methylation alterations were found at each of the myeloma stage in association with deregulated expression levels of Homeobox genes involved in osteogenic differentiation. Moreover, these DNA methylation changes were recapitulated in vitro by exposing MSCs from healthy individuals to MM plasma cells. Pharmacological targeting of DNMTs and G9a with the dual inhibitor CM-272, reverted the expression of aberrantly methylated osteogenic regulators and promoted OB differentiation of MSCs from myeloma patients. Most importantly, in a mouse model of bone marrow-disseminated MM, administration of CM-272 prevented tumor-associated bone loss and reduced tumor burden. Our results demonstrated that not only was aberrant DNA methylation a main contributor to bone formation impairment found in MM patients, but also its targeting by CM-272 was able to reverse MM-associated bone loss.

show abstract

“…The results of this study indicate that Btk enhanced the differentiation of osteoclasts in an inflammatory environment, which was mainly achieved by regulating the expression of PLC γ 2 and NFATc-1. Previous studies have also shown that in addition to the regulation of osteoclast differentiation, Btk can also regulate MAPK, NF- κ B, and PKC α signaling to suppress osteoblastic differentiation [31].…”

Section: Discussionmentioning

confidence: 99%

Role of the Btk-PLCγ2 Signaling Pathway in the Bone Destruction of Apical Periodontitis

Wang

Zhang

Dong

et al. 2019

Mediators of Inflammation

View full text Add to dashboard Cite

Chronic apical periodontitis is characterized by alveolar bone absorption in the apical region and is the result of the participation of various inflammatory mediators. Studies have shown that the Bruton tyrosine kinase- (Btk-) phospholipase Cγ2 (PLCγ2) signaling pathway plays an important role in bone absorption, but it is unknown whether it plays a role in apical periodontitis bone destruction. Therefore, this study verified the role of Btk and PLCγ2 in bone resorption of apical periodontitis by in vivo and in vitro experiments. In the in vivo experiment, a mice model of apical periodontitis was established; apical bone resorption was confirmed by the numbers of osteoclasts and HE staining. Btk, PLCγ2, and nuclear factor of activated T-cells 1 (NFATc-1) were detected by immunohistochemical staining. In the in vitro experiment, lipopolysaccharides (LPS) were used to stimulate osteoclast precursor cell RAW264.7 to establish an inflammatory microenvironment and detect osteoclast differentiation. By silencing Btk, the expression of Btk, PLCγ2, and NFATc-1 was detected by real-time qPCR and Western blot, and osteoclastogenesis was detected by enzyme histochemical staining to further confirm the role of Btk in bone resorption. It was found that the expression of Btk, PLCγ2, and NFATc-1 changed significantly with the progression of inflammation and bone destruction, indicating that Btk and PLCγ2 may be involved in the progression of inflammation in apical periodontitis and bone absorption. In vitro experiments confirmed that the differentiation of osteoclasts and the expression of PLCγ2 and NFATc-1 were significantly inhibited after silencing Btk expression, but osteoclast precursor cells could be differentiated due to the proinflammatory factor lipopolysaccharide. This study demonstrates that Btk and PLCγ2 are key factors involved in the apical inflammatory response and bone destruction.

show abstract

Bruton tyrosine kinase (Btk) suppresses osteoblastic differentiation

Cited by 4 publications

References 27 publications

Cell-Based Double-Screening Method to Identify a Reliable Candidate for Osteogenesis-Targeting Compounds

Cell-Based Double-Screening Method to Identify a Reliable Candidate for Osteogenesis-Targeting Compounds

Targeting aberrant DNA methylation in mesenchymal stromal cells as a treatment for myeloma bone disease

Role of the Btk-PLCγ2 Signaling Pathway in the Bone Destruction of Apical Periodontitis

Contact Info

Product

Resources

About