Stem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh, 11461, Kingdom of Saudi Arabia.
College of Medicine, Alfaisal University, Riyadh, 11533, Kingdom of Saudi Arabia.
Sci Rep. 2020 Oct 7;10(1):16746. doi: 10.1038/s41598-020-73439-9.
Tankyrase is part of poly (ADP-ribose) polymerase superfamily required for numerous cellular and molecular processes. Tankyrase inhibition negatively regulates Wnt pathway. Thus, Tankyrase inhibitors have been extensively investigated for the treatment of clinical conditions associated with activated Wnt signaling such as cancer and fibrotic diseases. Moreover, Tankyrase inhibition has been recently reported to upregulate osteogenesis through the accumulation of SH3 domain-binding protein 2, an adaptor protein required for bone metabolism. In this study, we investigated the effect of Tankyrase inhibition in osteoblast differentiation of human skeletal (mesenchymal) stem cells (hMSCs). A Tankyrase inhibitor, XAV-939, identified during a functional library screening of small molecules. Alkaline phosphatase activity and Alizarin red staining were employed as markers for osteoblastic differentiation and in vitro mineralized matrix formation, respectively. Global gene expression profiling was performed using the Agilent microarray platform. XAV-939, a Tankyrase inhibitor, enhanced osteoblast differentiation of hBMSCs as evidenced by increased ALP activity, in vitro mineralized matrix formation, and upregulation of osteoblast-related gene expression. Global gene expression profiling of XAV-939-treated cells identified 847 upregulated and 614 downregulated mRNA transcripts, compared to vehicle-treated control cells. It also points towards possible changes in multiple signaling pathways, including TGFβ, insulin signaling, focal adhesion, estrogen metabolism, oxidative stress, RANK-RANKL (receptor activator of nuclear factor κB ligand) signaling, Vitamin D synthesis, IL6, and cytokines and inflammatory responses. Further bioinformatic analysis, employing Ingenuity Pathway Analysis identified significant enrichment in XAV-939-treated cells of functional categories and networks involved in TNF, NFκB, and STAT signaling. We identified a Tankyrase inhibitor (XAV-939) as a powerful enhancer of osteoblastic differentiation of hBMSC that may be useful as a therapeutic option for treating conditions associated with low bone formation.
Tankyrase 属于多聚 ADP-核糖聚合酶超家族的一员，在许多细胞和分子过程中发挥作用。Tankyrase 的抑制作用可负向调节 Wnt 通路。因此，Tankyrase 抑制剂已被广泛研究用于治疗与激活的 Wnt 信号相关的临床病症，如癌症和纤维性疾病。此外，最近有报道称，Tankyrase 抑制作用可通过积累 SH3 结构域结合蛋白 2 来上调成骨作用，SH3 结构域结合蛋白 2 是一种参与骨代谢的衔接蛋白。在本研究中，我们研究了 Tankyrase 抑制剂对人成骨细胞（间充质）干细胞（hMSCs）成骨分化的影响。XAV-939 是在小分子功能文库筛选中发现的一种 Tankyrase 抑制剂。碱性磷酸酶活性和茜素红染色分别作为成骨分化和体外矿化基质形成的标志物。使用安捷伦微阵列平台进行全基因表达谱分析。Tankyrase 抑制剂 XAV-939 增强了 hBMSCs 的成骨分化，表现为碱性磷酸酶活性增加、体外矿化基质形成增加以及成骨相关基因表达上调。与对照组相比，XAV-939 处理细胞的全基因表达谱分析鉴定出 847 个上调和 614 个下调的 mRNA 转录本。这也表明可能发生了多种信号通路的变化，包括 TGFβ、胰岛素信号、焦点黏附、雌激素代谢、氧化应激、RANK-RANKL（核因子 κB 配体受体激活剂）信号、维生素 D 合成、IL6 和细胞因子及炎症反应。进一步的生物信息学分析，采用 Ingenuity Pathway Analysis 发现 XAV-939 处理细胞中显著富集了与 TNF、NFκB 和 STAT 信号相关的功能类别和网络。我们鉴定出一种 Tankyrase 抑制剂（XAV-939），它可增强 hBMSC 的成骨分化，可能是治疗与低骨形成相关疾病的一种有前途的治疗选择。
