ZNF281 Knockdown Induced Osteogenic Differentiation of Human Multipotent Stem Cells in Vivo and in Vitro

Abstract

ZNF281 is one of the core transcription factors in embryonic stem cells (ESCs) and has activation and repression roles in the transcription of ESC genes. A known target molecule of Zfp281 (the mouse homologue of ZNF281) is Nanog. However, NANOG is not expressed in most human multipotent stem cells (hMSCs). Here, we investigated the roles of ZNF281 with a gain- and loss-of-function study. The knockdown of ZNF281 in vivo and in vitro resulted in spontaneous osteochondrogenic differentiation and reduced the proliferation of hMSCs, as determined by cell morphology and molecular markers. When ZNF281-knockdown hMSCs were subcutaneously implanted into mice along with β-tricalcium phosphate (β-TCP), many cells were converted into osteoblasts within 4 weeks. In contrast, the overexpression of ZNF281 in hMSCs resulted in accelerated proliferation. The expression pattern of ZNF281 correlated well with the expression of β-CATENIN during differentiation and in the gain/loss-of-function study in hMSCs. The binding of ZNF281 to the promoter region of β-CATENIN was observed using a chromatin immunoprecipitation (ChIP) assay. In conclusion, we propose that ZNF281 plays an important role in the maintenance and osteogenic differentiation of stem cells via the transcriptional regulation of genes including β-CATENIN.

SNAIL and miR-34a feed-forward regulation of ZNF281/ZBP99 promotes epithelial-mesenchymal transition

“…However, ZNF281 may also promote β‐catenin/TCF4 activity by mediating the loss of E‐cadherin expression, which is known to inhibit nuclear localization and activity of β‐catenin by recruiting it to the cell membrane (Sadot et al, 1998; Orsulic et al, 1999). Furthermore, our results are consistent with the reported correlation of ZNF281 and β‐catenin expression in hMSCs (Seo et al, 2013). We found that ZNF281 induces expression of the prognostic stem‐cell markers CD133 and LGR5.…”

“…The enhancement of β‐catenin activity by ZNF281 may contribute to ZNF281‐induced stemness, since WNT signalling is necessary to maintain stem cells in the intestinal crypts (Pinto and Clevers, 2005; Brabletz et al, 2005b; Scoville et al, 2008). Interestingly, ZNF281 directly binds to the β ‐catenin promoter in human multipotent stem cells (hMSCs) (Seo et al, 2013). In addition, we found that ZNF281 directly represses Axin2 , a negative regulator of the WNT pathway (Lustig et al, 2002).…”

Transcription Factor ZBP-89 Drives a Feedforward Loop of β-Catenin Expression in Colorectal Cancer

“…Moreover, quantitative ChIP analysis revealed that ZBP-99 occupies the promoter of the Wnt target gene LGR5 , which ultimately increases β-catenin activity and the acquisition of stem cell traits (38). ZBP-99 binds the proximal CTNNB1 promoter and induces CTNNB1 mRNA in osteogenic stem cells, demonstrating that a ZBP-89 family member directly binds and regulates CTNNB1 gene expression (40). Thus, ZBP-99, a ZBP-89 family member, is able to indirectly regulate β-catenin activity in CRC cell lines, and directly bind to the CTNNB1 promoter in specific cell types, suggesting that ZBP-89 might also regulate CTNNB1 gene expression.…”

MicroRNA-188 regulates age-related switch between osteoblast and adipocyte differentiation

“…We used miRanda (19), PicTar (20), and TargetScan (21) to predict the targets of miR-188. Among the predicted genes, we chose 6 for further analysis-histone deacetylase 9 (Hdac9), RPTORindependent companion of MTOR complex 2 (Rictor), phosphatase and tensin homolog (Pten), zinc finger protein 281 (Znf281), GLIS family zinc finger 3 (Glis3), and ephrin B2 (Efnb2)-which had been reported to participate in bone metabolism (22)(23)(24)(25)(26)(27)(28). Overexpression or inhibition of miR-188 changed endogenous levels of HDAC9 and RICTOR protein, but not the others ( Figure 8A).…”