Transcriptional regulation of FRZB in chondrocytes by Osterix and Msx2
Transcriptional regulation of FRZB in chondrocytes by Osterix and Msx2
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DOI：
10.1007/s00774-022-01345-3
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发表时间：
2022-06-28
影响因子：
3.3
通讯作者：
Nishimura，Riko
中科院分区：
文献类型：
--
作者：
Yagi，Hiroko;Takahata，Yoshifumi;Nishimura，Riko
骨关节炎是一种常见的关节疾病，可导致膝关节和髋关节周围的关节软骨破坏和严重炎症。然而，迄今为止，由于潜在的分子机制复杂，骨关节炎的有效治疗试剂尚未开发出来。最近的遗传学发现表明，Wnt拮抗剂卷曲相关蛋白B （FRZB）是骨关节炎的潜在治疗靶点。因此，本研究旨在研究rzbin软骨细胞的转录调控。材料与方法采用RT-qPCR方法检测小鼠关节软骨细胞和SW1353软骨细胞系中frzb / frzb的表达。用腺病毒和慢病毒分别进行过表达和敲低实验。荧光素酶报告和染色质免疫沉淀法测定转录调节。蛋白-蛋白相互作用通过共免疫沉淀分析确定。结果frzb蛋白在软骨中高度表达，尤其是在关节软骨细胞中。白细胞介素-1α显著降低了与软骨破坏相关的关节软骨细胞中frz的表达，并增加了带有血小板反应蛋白1型基序（Adamts） 4和adamts5表达的ADAM金属肽酶。骨形态发生蛋白2 （Bone morphogenetic protein 2, BMP2）通过Smad信号增加SW1353细胞中frzbe1的表达。Osterix和msh homeobox 2 （Msx2）都是BMP2的下游转录因子，它们诱导frzbe1的表达并上调其启动子活性。共免疫沉淀结果显示Osterix与Msx2之间存在物理相互作用。敲低Osterix或Msx2均可抑制bmp2依赖性frzbein的表达。染色质免疫沉淀显示Osterix和Msx2与frzb基因启动子直接相关。结论BMP2通过Osterix和Msx2调控frzbein的表达。
IntroductionOsteoarthritis is a common joint disease that causes destruction of articular cartilage and severe inflammation surrounding knee and hip joints. However, to date, effective therapeutic reagents for osteoarthritis have not been developed because the underlying molecular mechanisms are complex. Recent genetic findings suggest that a Wnt antagonist, frizzled-related protein B (FRZB), is a potential therapeutic target for osteoarthritis. Therefore, this study aimed to examine the transcriptional regulation ofFRZBin chondrocytes.Materials and methodsFrzb/FRZBexpression was assessed by RT-qPCR analyses in murine articular chondrocytes and SW1353 chondrocyte cell line. Overexpression and knockdown experiments were performed using adenovirus and lentivirus, respectively. Luciferase-reporter and chromatin immunoprecipitation assays were performed for determining transcriptional regulation. Protein–protein interaction was determined by co-immunoprecipitation analysis.ResultsFrzbwas highly expressed in cartilages, especially within articular chondrocytes. Interleukin-1α markedly reducedFrzbexpression in articular chondrocytes in association with cartilage destruction and increases in ADAM metallopeptidase with thrombospondin type 1 motif (Adamts) 4 andAdamts5expression. Bone morphogenetic protein 2 (BMP2) increasedFRZBexpression in SW1353 cells through Smad signaling. Osterix and msh homeobox 2 (Msx2), both of which function as downstream transcription factors of BMP2, inducedFRZBexpression and upregulated its promoter activity. Co-immunoprecipitation results showed a physical interaction between Osterix and Msx2. Knockdown of either Osterix or Msx2 inhibited BMP2-dependentFRZBexpression. Chromatin immunoprecipitation indicated a direct association of Osterix and Msx2 with theFRZBgene promoter.ConclusionThese results suggest that BMP2 regulatesFRZBexpression through Osterix and Msx2.