Abstract
Yes-associated protein 1 (YAP1) is a transcriptional co-activator that is involved in cell proliferation and differentiation. The aim of this study is to investigate the effect of inhibition of YAP1 activity with vertepofin on the differentiation of mesenchymal cells at different time-points. Inhibition of YAP1 from the beginning of culture with verteporfin resulted in the suppression of chondrogenesis and cell aggregation, as well as expression of N-cadherin and fibronectin. Culturing constitutively active YAP1-overexpressing cells for 3 days had no effect on cell aggregation and differentiation and increased the expression of N-cadherin, while culturing these cells for 7 days displayed decreased Alcian blue staining. Inhibition of YAP1 activity with verteporfin at the late stage of each culture with different culture periods did not affect chondrogenesis. When the medium of cells cultured for 3, 6, or 8 days was changed to medium containing verteporfin, no changes in Alcian blue staining were evident in the next 1 to 2 days. Interestingly, in cells grown for 8 days in normal medium and incubated for 2 more days in the presence of verteporfin, Alizarin red S staining and osteopontin expression increased. Collectively, our results revealed that verteporfin regulates cell aggregation during early-stage differentiation and promotes osteogenesis at a late stage by inhibiting YAP1 activity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ahrens PB, Solursh M, Reiter RS (1977) Stage-related capacity for limb chondrogenesis in cell culture. Dev Biol 60:69–82
Aragona M, Panciera T, Manfrin A, Giulitti S, Michielin F, Elvassore N, Dupont S, Piccolo S (2013) A mechanical checkpoint controls multicellular growth through YAP/TAZ regulation by actin-processing factors. Cell 154:1047–1059. https://doi.org/10.1016/j.cell.2013.07.042
Blom AB, van Lent PL, van der Kraan PM, van den Berg WB (2010) To seek shelter from the Wnt in osteoarthritis? Wnt-signaling as a target for osteoarthritis therapy. Curr Drug Targets 11:620–629
DeLise AM, Tuan RS (2002) Analysis of N-cadherin function in limb mesenchymal chondrogenesis in vitro. Dev Dyn 225:195–204
DeLise AM, Fischer L, Tuan RS (2000) Cellular interactions and signaling in cartilage development. Osteoarthr Cartil 8:309–334
Deng Y, Wu A, Li P, Li G, Qin L, Song H, Mak KK (2016) Yap1 regulates multiple steps of chondrocyte differentiation during skeletal development and bone repair. Cell Rep 14:2224–2237. https://doi.org/10.1016/j.celrep.2016.02.021
Deng F, Peng L, Li Z, Tan G, Liang E, Chen S, Zhao X, Zhi F (2018) YAP triggers the Wnt/β-catenin signalling pathway and promotes enterocyte self-renewal, regeneration and tumorigenesis after DSS-induced injury. Cell Death Dis 9:153
Dessau W, Von der Mark H, Von der Mark K, Fischer S (1980) Changes in the patterns of collagens and fibronectin during limb-bud chondrogenesis. J Embryol Exp Morphol 57:51–60
Dupont S, Morsut L, Aragona M, Enzo E, Giulitti S, Cordenonsi M, Zanconato F, Le Digabel J, Forcato M, Bicciato S, Elvassore N, Piccolo S (2011) Role of YAP/TAZ in mechanotransduction. Nature 474:179–183. https://doi.org/10.1038/nature10137
Eales KL, Wilkinson EA, Cruickshank G, Tucker JHR, Tennant DA (2018) Verteporfin selectively kills hypoxic glioma cells through iron-binding and increased production of reactive oxygen species. Sci Rep 8:14358. https://doi.org/10.1038/s41598-018-32727-1
Funck-Brentano T, Bouaziz W, Marty C, Geoffroy V, Hay E, Cohen-Solal M (2014) Dkk-1-mediated inhibition of Wnt signaling in bone ameliorates osteoarthritis in mice. Arthritis Rheumatol 66:3028–3039. https://doi.org/10.1002/art.38799
Halder G, Johnson RL (2011) Hippo signaling: growth control and beyond. Development 138:9–22. https://doi.org/10.1242/dev.045500
Hall BK, Miyake T (2000) All for one and one for all: condensations and the initiation of skeletal development. Bioessays 22:138–147
Imajo M, Miyatake K, Iimura A, Miyamoto A, Nishida E (2012) A molecular mechanism that links Hippo signalling to the inhibition of Wnt/β-catenin signalling. EMBO J 31:1109–1122. https://doi.org/10.1038/emboj.2011.487
Judson RN, Tremblay AM, Knopp P, White RB, Urcia R, De Bari C, Zammit PS, Camargo FD, Wackerhage H (2012) The Hippo pathway member Yap plays a key role in influencing fate decisions in muscle satellite cells. J Cell Sci 125:6009–6019. https://doi.org/10.1242/jcs.109546
Kahles F, Findeisen HM, Bruemmer D (2014) Osteopontin: a novel regulator at the cross roads of inflammation, obesity and diabetes. Mol Metab 3:384–393. https://doi.org/10.1016/j.molmet.2014.03.004
Karystinou A, Roelofs AJ, Neve A, Cantatore FP, Wackerhage H, De Bari C (2015) Yes-associated protein (YAP) is a negative regulator of chondrogenesis in mesenchymal stem cells. Arthritis Res Ther 17:147. https://doi.org/10.1186/s13075-015-0639-9
Konsavage WM Jr, Yochum GS (2013) Intersection of Hippo/YAP and Wnt/β-catenin signaling pathways. Acta Biochim Biophys Sin Shanghai 45:71–79. https://doi.org/10.1093/abbs/gms084
Konsavage WM Jr, Kyler SL, Rennoll SA, Jin G, Yochum GS (2012) Wnt/β-catenin signaling regulates Yes-associated protein (YAP) gene expression in colorectal carcinoma cells. J Biol Chem 287:11730–11739. https://doi.org/10.1074/jbc.M111.327767
Kulyk WM, Upholt WB, Kosher RA (1989) Fibronectin gene expression during limb cartilage differentiation. Development 106:449–455
Lietman C, Wu B, Lechner S, Shinar A, Sehgal M, Rossomacha E, Datta P, Sharma A, Gandhi R, Kapoor M, Young PP (2018) Inhibition of Wnt/β-catenin signaling ameliorates osteoarthritis in a murine model of experimental osteoarthritis. JCI Insight 96308. https://doi.org/10.1172/jci.insight.96308
Liu-Chittenden Y, Huang B, Shim JS, Chen Q, Lee SJ, Anders RA, Liu JO, Pan D (2012) Genetic and pharmacological disruption of the TEAD-YAP complex suppresses the oncogenic activity of YAP. Genes Dev 26:1300–1305
Luyten FP, Tylzanowski P, Lories RJ (2009) Wnt signaling and osteoarthritis. Bone 44:522–527. https://doi.org/10.1016/j.bone.2008.12.006
Piccolo S, Dupont S, Cordenonsi M (2014) The biology of YAP/TAZ: hippo signaling and beyond. Physiol Rev 94:1287–1312. https://doi.org/10.1152/physrev.00005.2014
Ranscht B (1994) Cadherins and catenins: interactions and functions in embryonic development. Curr Opin Cell Biol 6:740–746
Ren F, Zhang L, Jiang J (2010) Hippo signaling regulates Yorkie nuclear localization and activity through 14-3-3 dependent and independent mechanisms. Dev Biol 337:303–312. https://doi.org/10.1016/j.ydbio.2009.10.046
Singh P, Schwarzbauer JE (2014) Fibronectin matrix assembly is essential for cell condensation during chondrogenesis. J Cell Sci 127:4420–4428. https://doi.org/10.1242/jcs.095786
Takács R, Matta C, Somogyi C, Juhász T, Zákány R (2013) Comparative analysis of osteogenic/chondrogenic differentiation potential in primary limb bud-derived and C3H10T1/2 cell line-based mouse micromass cultures. Int J Mol Sci 14:16141–16167. https://doi.org/10.3390/ijms140816141
Wang C, Zhu X, Feng W, Yu Y, Jeong K, Guo W, Lu Y, Mills GB (2016) Verteporfin inhibits YAP function through up-regulating 14-3-3σ sequestering YAP in the cytoplasm. Am J Cancer Res 6:27–37
Wei H, Wang F, Wang Y, Li T, Xiu P, Zhong J, Sun X, Li J (2017) Verteporfin suppresses cell survival, angiogenesis and vasculogenic mimicry of pancreatic ductal adenocarcinoma via disrupting the YAP-TEAD complex. Cancer Sci 108:478–487. https://doi.org/10.1111/cas.13138
Yu FX, Zhao B, Panupinthu N, Jewell JL, Lian I, Wang LH, Zhao J, Yuan H, Tumaneng K, Li H, Fu XD, Mills GB, Guan KL (2012) Regulation of the Hippo-YAP pathway by G-protein-coupled receptor signaling. Cell 150:780–791. https://doi.org/10.1016/j.cell.2012.06.037
Zanconato F, Forcato M, Battilana G, Azzolin L, Quaranta E, Bodega B, Rosato A, Bicciato S, Cordenonsi M, Piccolo S (2015) Genome-wide association between YAP/TAZ/TEAD and AP-1 at enhancers drives oncogenic growth. Nat Cell Biol 17:1218–1227. https://doi.org/10.1038/ncb3216
Zhang X, Ziran N, Goater JJ, Schwarz EM, Puzas JE, Rosier RN, Zuscik M, Drissi H, O'Keefe RJ (2004) Primary murine limb bud mesenchymal cells in long-term culture complete chondrocyte differentiation: TGF-beta delays hypertrophy and PGE2 inhibits terminal differentiation. Bone 34:809–817
Zhang W, Ouyang H, Dass CR, Xu J (2016) Current research on pharmacologic and regenerative therapies for osteoarthritis. Bone Res 4:15040. https://doi.org/10.1038/boneres.2015.40
Zhao B, Wei X, Li W, Udan RS, Yang Q, Kim J, Xie J, Ikenoue T, Yu J, Li L, Zheng P, Ye K, Chinnaiyan A, Halder G, Lai ZC, Guan KL (2007) Inactivation of YAP oncoprotein by the hippo pathway is involved in cell contact inhibition and tissue growth control. Genes Dev 21:2747–2761
Zhao B, Lei QY, Guan KL (2008a) The Hippo-YAP pathway: new connections between regulation of organ size and cancer. Curr Opin Cell Biol 20:638–646. https://doi.org/10.1016/j.ceb.2008.10.001
Zhao B, Ye X, Yu J, Li L, Li W, Li S, Yu J, Lin JD, Wang CY, Chinnaiyan AM, Lai ZC, Guan KL (2008b) TEAD mediates YAP-dependent gene induction and growth control. Genes Dev 22:1962–1971. https://doi.org/10.1101/gad.1664408
Zhong W, Li Y, Li L, Zhang W, Wang S, Zheng X (2013) YAP-mediated regulation of the chondrogenic phenotype in response to matrix elasticity. J Mol Histol 44:587–595. https://doi.org/10.1007/s10735-013-9502-y
Zollinger AJ, Smith ML (2017) Fibronectin, the extracellular glue. Matrix Biol 60-61:27–37. https://doi.org/10.1016/j.matbio.2016.07.011
Acknowledgements
This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education [2017R1D1A3B03027848].
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Jeong, B., Kim, D., Kim, J. et al. Stage differential effects of verteporfin on the differentiation of chick embryo wing bud mesenchymal cells. Biologia 74, 1219–1228 (2019). https://doi.org/10.2478/s11756-019-00282-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.2478/s11756-019-00282-3