Tankyrase inhibitor XAV-939 enhances osteoblastogenesis and mineralization of human skeletal (mesenchymal) stem cells

Abstract

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.

Pharmacological Manipulation of Wnt/β-Catenin Signaling Pathway in Human Neural Precursor Cells Alters Their Differentiation Potential and Neuronal Yield

“…Taking advantage of our in vitro drug screening platform of self-renewable hNPCs, we also asked whether CHIR or XAV might be cytotoxic, by measuring cell proliferation and survival. Both drugs were used in this study at the same concentration or lower as in similar studies employing human or murine ESCs, hiPSCs and other types of stem cells ( De Kumar et al, 2017 ; Malleske et al, 2018 ; Shafa et al, 2018 ; Gomez et al, 2019 ; Hamad et al, 2019 ; Qiu et al, 2019 ; Shin et al, 2019 ; Almasoud et al, 2020 ; Bataille et al, 2020 ; Bejoy et al, 2020 ; Govarthanan et al, 2020 ; Han et al, 2020 ; Kanagaki et al, 2020 ; Leigh et al, 2020 ; Yang J. et al, 2020 ; Yang Y. et al, 2020 ; Ren et al, 2021 ; Wang et al, 2021 ). When stem cells differentiate into more mature cell types, their proliferation potential is gradually reduced, until cells reach a post-mitotic fate.…”

“…CHIR has been used to promote neural and neuronal differentiation in embryonic stem cells and induced pluripotent stem cells ( De Kumar et al, 2017 ; Shafa et al, 2018 ; Gomez et al, 2019 ; Qiu et al, 2019 ; Shin et al, 2019 ; Bejoy et al, 2020 ) and in several different types of neural precursors ( Yang Y. et al, 2020 ; Ren et al, 2021 ; Wang et al, 2021 ), as well as to induce trans- differentiation into neural lineages from non-neural cell types, such as skin ( Bataille et al, 2020 ; Yang J. et al, 2020 ) and mesenchymal cells ( Govarthanan et al, 2020 ). XAV has been used to promote differentiation of pluripotent stem cells into lung cells ( Malleske et al, 2018 ; Kanagaki et al, 2020 ), to increase osteogenic differentiation of mesenchymal stem cells ( Almasoud et al, 2020 ; Han et al, 2020 ), and to promote generation of cardiomyocytes ( Hamad et al, 2019 ; Leigh et al, 2020 ).…”

Possible Action of Olaparib for Preventing Invasion of Oral Squamous Cell Carcinoma In Vitro and In Vivo

“…Regarding osteoblastogenesis, a critical role of PARP1 was also previously reported. Our previous study demonstrated that PARP inhibitor PJ34 attenuated mesenchymal osteoblast differentiation via the BMP-2 signaling pathway in vitro [ 53 ], which suggested a progressive effect on osteoclastogenesis; furthermore, another study showed that tankyrase inhibitor XAV-939 enhances osteoblastogenesis and mineralization of human skeletal mesenchymal stem cells via TNF, NF-κB, and STAT signaling [ 54 ]. Thus, the effect of olaparib is suggested to be pivotal on other pathways, or this role could be due to the differences in the derived cell lines.…”

JAK2 Inhibition by Fedratinib Reduces Osteoblast Differentiation and Mineralisation of Human Mesenchymal Stem Cells

“…Small molecule inhibitors targeting specific intracellular signalling pathways are currently used in vitro as a biochemical tool to identify the molecular mechanisms involved in stem cell differentiation into osteoblast lineages [ 15 , 20 , 22 , 24 , 25 , 26 , 27 ]. Here, we identified a small molecule Fedratinib, a potent JAK2 inhibitor [ 28 ], through a small molecule library screen [ 15 ], as a powerful inhibitor of osteoblastic differentiation of hMSC-TERT cells.…”