ZBP1 (DAI/DLM-1) promotes osteogenic differentiation while inhibiting adipogenic differentiation in mesenchymal stem cells through a positive feedback loop of Wnt/β-catenin signaling

Abstract

The lineage specification of mesenchymal stem/stromal cells (MSCs) is tightly regulated by a wide range of factors. Recently, the versatile functions of ZBP1 (also known as DAI or DLM-1) have been reported in the blood circulation and immune systems. However, the biological function of ZBP1 during the lineage specification of MSCs is still unknown. In the present study, we found that ZBP1 was upregulated during osteogenesis but downregulated during adipogenesis in mouse bone marrow-derived MSCs (mBMSCs). ZBP1 was highly expressed in osteoblasts but expressed at a relatively low level in marrow adipocytes. Knockdown of ZBP1 inhibited alkaline phosphataseactivity, extracellular matrix mineralization, and osteogenesis-related gene expression in vitro and reduced ectopic bone formation in vivo. Knockdown of ZBP1 also promoted adipogenesis in MSCs in vitro. Conversely, the overexpression of ZBP1 increased the osteogenesis but suppressed the adipogenesis of MSCs. When the expression of ZBP1 was rescued, the osteogenic capacity of ZBP1-depleted mBMSCs was restored at both the molecular and phenotypic levels. Furthermore, we demonstrated that ZBP1, a newly identified target of Wnt/β-catenin signaling, was required for β-catenin translocation into nuclei. Collectively, our results indicate that ZBP1 is a novel regulator of bone and fat transdifferentiation via Wnt/β-catenin signaling.

References

[1] Nombela-Arrieta C, Ritz J, Silberstein LE. The elusive nature and function of mesenchymal stem cells. Mol. Cell Biol., 2011, 12:126-131

[2] Koc ON et al. Bone marrow-derived mesenchymal stem cells remain host-derived despite successful hematopoietic engraftment after allogeneic transplantation in patients with lysosomal and peroxisomal storage diseases. Exp. Hematol., 1999, 27:1675-1681

[3] Lee OK et al. Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood, 2004, 103:1669-1675

[4] Lee RH et al. Characterization and expression analysis of mesenchymal stem cells from human bone marrow and adipose tissue. Cell Physiol. Biochem., 2004, 14:311-324

[5] Kim N, Cho SG. Clinical applications of mesenchymal stem cells. Korean J. Intern. Med., 2013, 28:387-402

[6] Wang S, Qu X, Zhao RC. Clinical applications of mesenchymal stem cells. J. Hematol. Oncol., 2012, 5:19

[7] Neve A, Corrado A, Cantatore FP. Osteoblast physiology in normal and pathological conditions. Cell Tissue Res., 2011, 343:289-302

[8] Deng P, Chen Q-M, Hong C, Wang C-Y. Histone methyltransferases and demethylases: regulators in balancing osteogenic and adipogenic differentiation of mesenchymal stem cells. Int. J. Oral. Sci., 2015, 7:197-204

[9] Liu W et al. GDF11 decreases bone mass by stimulating osteoclastogenesis and inhibiting osteoblast differentiation. Nat. Commun., 2016, 7

[10] Krishnan V, Bryant HU, MacDougald OA. Regulation of bone mass by Wnt signaling. J. Clin. Investig., 2006, 116:1202-1209

[11] Hay E et al. Interaction between LRP5 and Frat1 mediates the activation of the wnt canonical pathway. J. Biol. Chem., 2005, 280:13616-13623

[12] Levy L., Wei Y., Labalette C., Wu Y., Renard C.-A., Buendia M. A., Neuveut C.. Acetylation of  -Catenin by p300 Regulates  -Catenin-Tcf4 Interaction. Molecular and Cellular Biology, 2004, 24 8 3404-3414

[13] Bennett CN et al. Regulation of osteoblastogenesis and bone mass by Wnt10b. Proc. Natl Acad. Sci. USA, 2005, 102:3324-3329

[14] Jackson A et al. Gene array analysis of Wnt-regulated genes in C3H10T1/2 cells. Bone, 2005, 36:585-598

[15] Gaur T et al. Canonical WNT signaling promotes osteogenesis by directly stimulating Runx2 gene expression. J. Biol. Chem., 2005, 280:33132-33140

[16] Rodríguez-Carballo E et al. Conserved regulatory motifs in osteogenic gene promoters integrate cooperative effects of canonical Wnt and BMP pathways. J. Bone Miner. Res., 2011, 26:718-729

[17] Liu B, Wu S, Han L, Zhang C. beta-catenin signaling induces the osteoblastogenic differentiation of human pre-osteoblastic and bone marrow stromal cells mainly through the upregulation of osterix expression. Int. J. Mol. Med., 2015, 36:1572-1582

[18] Ross SE et al. Inhibition of adipogenesis by Wnt signaling. Science, 2000, 289:950-953

[19] Day TF, Guo X, Garrett-Beal L, Yang Y. Wnt/beta-catenin signaling in mesenchymal progenitors controls osteoblast and chondrocyte differentiation during vertebrate skeletogenesis. Dev. Cell, 2005, 8:739-750

[20] Schwartz T, Behlke J, Lowenhaupt K, Heinemann U, Rich A. Structure of the DLM-1–Z-DNA complex reveals a conserved family of Z-DNA-binding proteins. Nat. Struct. Biol., 2001, 8:761-765

[21] Fu Y et al. Cloning of DLM-1, a novel gene that is up-regulated in activated macrophages, using RNA differential display. Gene, 1999, 240:157-163

[22] Rothenburg S, Schwartz T, Koch-Nolte F, Haag F. Complex regulation of the human gene for the Z-DNA binding protein DLM-1. Nucleic Acids Res., 2002, 30:993-1000

[23] Yue F et al. A comparative encyclopedia of DNA elements in the mouse genome. Nature, 2014, 515:355-364

[24] Takaoka A et al. DAI (DLM-1/ZBP1) is a cytosolic DNA sensor and an activator of innate immune response. Nature, 2007, 448:501-505

[25] Kesavardhana S et al. ZBP1/DAI ubiquitination and sensing of influenza vRNPs activate programmed cell death. J. Exp. Med, 2017, 214:12

[26] Huang DW, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat. Protoc., 2008, 4:44

[27] Subramanian A et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl Acad. Sci. USA, 2005, 102:15545-15550

[28] Mathelier A et al. JASPAR 2016: a major expansion and update of the open-access database of transcription factor binding profiles. Nucleic Acids Res., 2016, 44:D110-D115

[29] Zhao X et al. Cysteine dioxygenase type 1 inhibits osteogenesis by regulating Wnt signaling in primary mouse bone marrow stromal cells. Sci. Rep., 2016, 6

[30] Bianco P et al. The meaning, the sense and the significance: translating the science of mesenchymal stem cells into medicine. Nat. Med., 2013, 19:35-42

[31] DeFilippis VR, Alvarado D, Sali T, Rothenburg S, Früh K. Human cytomegalovirus induces the interferon response via the DNA sensor ZBP1. J. Virol., 2010, 84:585-598

[32] Kuriakose T, Kanneganti T-D. ZBP1: innate sensor regulating cell death and inflammation. Trends Immunol., 2018, 39:123-134

[33] Beresford JN, Bennett JH, Devlin C, Leboy PS, Owen ME. Evidence for an inverse relationship between the differentiation of adipocytic and osteogenic cells in rat marrow stromal cell cultures. J. Cell Sci., 1992, 102:341-351

[34] Rebsamen M et al. DAI/ZBP1 recruits RIP1 and RIP3 through RIP homotypic interaction motifs to activate NF-kappaB. EMBO Rep., 2009, 10:916-922

[35] Jonason JH, Xiao G, Zhang M, Xing L, Chen D. Post-translational regulation of Runx2 in bone and cartilage. J. Dent. Res., 2009, 88:693-703

[36] Nakashima K et al. The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell, 2002, 108:17-29

[37] Wölfl S, Wittig B, Dorbic T, Rich A. Identification of processes that influence negative supercoiling in the human c-myc gene. Biochim. Biophys. Acta, Gene Struct. Expr., 1997, 1352:213-221

[38] Wittig B, Wölfl S, Dorbic T, Vahrson W, Rich A. Transcription of human c-myc in permeabilized nuclei is associated with formation of Z-DNA in three discrete regions of the gene. EMBO J., 1992, 11:4653-4663

[39] Wölfl S, Martinez C, Rich A, Majzoub JA. Transcription of the human corticotropin-releasing hormone gene in NPLC cells is correlated with Z-DNA formation. Proc. Nat. Acad. Sci. USA, 1996, 93:3664-3668

[40] Oh DB, Kim YG, Rich A. Z-DNA-binding proteins can act as potent effectors of gene expression in vivo. Proc. Nat. Acad. Sci. USA, 2002, 99:16666-16671

[41] Soleimani M, Nadri S. A protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow. Nat. Protoc., 2009, 4:102-106

[42] Deng P et al. Cysteine dioxygenase type 1 promotes adipogenesis via interaction with peroxisome proliferator-activated receptor gamma. Biochem. Biophys. Res. Commun., 2015, 458:123-127

[43] Detre S, Saclani Jotti G, Dowsett M. A “quickscore” method for immunohistochemical semiquantitation: validation for oestrogen receptor in breast carcinomas. J. Clin. Pathol., 1995, 48:876-878

[44] Deng P et al. AFF4 promotes tumorigenesis and tumor-initiation capacity of head and neck squamous cell carcinoma cells by regulating SOX2. Carcinogenesis, 2018, 39:937-947

[45] Li J et al. KDM3 epigenetically controls tumorigenic potentials of human colorectal cancer stem cells through Wnt/β-catenin signalling. Nat. Commun., 2017, 8

[46] Deng P, Zhou C, Alvarez R, Hong C, Wang CY. Inhibition of IKK/NF-kappaB signaling enhances differentiation of mesenchymal stromal cells from human embryonic stem cells. Stem Cell Rep., 2016, 6:456-465

Funding

National Natural Science Foundation of China (National Science Foundation of China)(81600877)

China Postdoctoral Science Foundation(2016M600745)