Regulation of tumor angiogenesis by the microtubule-binding protein CLIP-170

Abstract

Angiogenesis, the expansion of preexisting blood vessels, is a complex process required for tumor growth and metastasis. Although current antiangiogenic strategies have shown promising results in several cancer types, identifi-cation of additional antiangiogenic targets is required to improve the therapeutic response. Herein, we show that the microtubule-binding protein CLIP-170 (cytoplasmic linker protein of 170 kDa) is highly expressed in breast tumor samples and correlates positively with blood vessel density. Depletion of CLIP-170 significantly impaired vascular endothelial tube formation and sprouting in vitro and inhibited breast tumor growth in mice by decreasing tumor vascularization. Our data further show that CLIP-170 is important for the migration but not the proliferation of vascular endothelial cells. In addition, CLIP-170 promotes the polarization of endothelial cells in response to the angiogenic stimulus. These findings thus demonstrate a critical role for CLIP-170 in tumor angiogenesis and suggest its potential as a novel antiangiogenic target

Key words： tumor angiogenesis; endothelial cell; migration; polarization; microtubule

References

[1] Akhmanova, A., and Steinmetz, M.O. (2008). Tracking the ends: a dynamic protein network controls the fate of microtubule tips. Nat Rev Mol Cell Biol 9, 309-322 .10.1038/nrm2369
[2] Bilbe, G., Delabie, J., Bruggen, J., Richener, H., Asselbergs, F.A., Cerletti, N., Sorg, C., Odink, K., Tarcsay, L., Wiesendanger, W., . (1992). Restin: a novel intermediate filament-associated protein highly expressed in the Reed-Sternberg cells of Hodgkin’s disease. EMBO J 11, 2103-2113 .
[3] Bornens, M. (2008). Organelle positioning and cell polarity. Nat Rev Mol Cell Biol 9, 874-886 .10.1038/nrm2524
[4] Bossung, V., and Harbeck, N. (2010). Angiogenesis inhibitors in the management of breast cancer. Curr Opin Obstet Gynecol 22, 79-86 .10.1097/GCO.0b013e328334e462
[5] Carmeliet, P., and Jain, R.K. (2011). Molecular mechanisms and clinical applications of angiogenesis. Nature 473, 298-307 .10.1038/nature10144
[6] Chae, S.S., Paik, J.H., Furneaux, H., and Hla, T. (2004). Requirement for sphingosine 1-phosphate receptor-1 in tumor angiogenesis demonstrated by in vivo RNA interference. J Clin Invest 114, 1082-1089 .
[7] Delabie, J., Shipman, R., Bruggen, J., De Strooper, B.,van Leuven, F., Tarcsay, L., Cerletti, N., Odink, K., Diehl, V., Bilbe, G., . (1992). Expression of the novel intermediate filament-associated protein restin in Hodgkin’s disease and anaplastic large-cell lymphoma. Blood 80, 2891-2896 .
[8] Dong, X., Liu, F., Sun, L., Liu, M., Li, D., Su, D., Zhu, Z., Dong, J.T., Fu, L., and Zhou, J. (2010). Oncogenic function of microtubule endbinding protein 1 in breast cancer. J Pathol 220, 361-369 .
[9] Folkman, J. (2007). Angiogenesis: an organizing principle for drug dis-covery? Nat Rev Drug Discov 6, 273-286 .10.1038/nrd2115
[10] Gao, J., Sun, L., Huo, L., Liu, M., Li, D., and Zhou, J. (2010). CYLD regulates angiogenesis by mediating vascular endothelial cell migration. Blood 115, 4130-4137 .10.1182/blood-2009-10-248526
[11] Gouveia, S.M., and Akhmanova, A. (2010). Cell and molecular biology of microtubule plus end tracking proteins: end binding proteins and their partners. Int Rev Cell Mol Biol 285, 1-74 .10.1016/B978-0-12-381047-2.00001-3
[12] Lamalice, L., Le Boeuf, F., and Huot, J. (2007). Endothelial cell migration during angiogenesis. Circ Res 100, 782-794 .10.1161/01.RES.0000259593.07661.1e
[13] Li, D., Xie, S., Ren, Y., Huo, L., Gao, J., Cui, D., Liu, M., and Zhou, J. (2011). Microtubule-associated deacetylase HDAC6 promotes angiogenesis by regulating cell migration in an EB1-dependent manner. Protein Cell 2, 150-160 .10.1007/s13238-011-1015-4
[14] Li, J., Sun, X., Wang, Z., Chen, L., Li, D., Zhou, J., and Liu, M. (2012). Regulation of vascular endothelial cell polarization and migration by Hsp70/Hsp90-organizing protein. PLoS One 7, e36389.10.1371/journal.pone.0036389
[15] Li, R., and Gundersen, G.G. (2008). Beyond polymer polarity: how the cytoskeleton builds a polarized cell. Nat Rev Mol Cell Biol 9, 860-873 .10.1038/nrm2522
[16] Liu, M., Wang, X., Yang, Y., Li, D., Ren, H., Zhu, Q., Chen, Q., Han, S., Hao, J., and Zhou, J. (2010). Ectopic expression of the microtubuledependent motor protein Eg5 promotes pancreatic tumourigenesis. J Pathol 221, 221-228 .10.1002/path.2706
[17] Mackey, J.R., Kerbel, R.S., Gelmon, K.A., McLeod, D.M., Chia, S.K., Rayson, D., Verma, S., Collins, L.L., Paterson, A.H., Robidoux, A., . (2012). Controlling angiogenesis in breast cancer: a systematic review of anti-angiogenic trials. Cancer Treat Rev 38, 673-688 .10.1016/j.ctrv.2011.12.002
[18] Peng, G., Ren, Y., Sun, X., Zhou, J., and Li, D. (2012). Inhibition of farnesyltransferase reduces angiogenesis by interrupting endothe lial cell migration. Biochem Pharmacol 83, 1374-1382 .10.1016/j.bcp.2012.02.008
[19] Rodriguez, O.C., Schaefer, A.W., Mandato, C.A., Forscher, P., Bement, W.M., and Waterman-Storer, C.M. (2003). Conserved microtubuleactin interactions in cell movement and morphogenesis. Nat Cell Biol 5, 599-609 .10.1038/ncb0703-599
[20] Shi, X., Liu, M., Li, D., Wang, J., Aneja, R., and Zhou, J. (2012). Cep70 contributes to angiogenesis by modulating microtubule rearrangement and stimulating cell polarization and migration. Cell Cycle 11, 1554-1563 .10.4161/cc.19954
[21] Sun, L., Gao, J., Dong, X., Liu, M., Li, D., Shi, X., Dong, J.T., Lu, X., Liu, C., and Zhou, J. (2008). EB1 promotes Aurora-B kinase activity through blocking its inactivation by protein phosphatase 2A. Proc Natl Acad Sci U S A 105, 7153-7158 .10.1073/pnas.0710018105
[22] Sun, X., Li, D., Yang, Y., Ren, Y., Li, J., Wang, Z., Dong, B., Liu, M., and Zhou, J. (2012). Microtubule-binding protein CLIP-170 is a mediator of paclitaxel sensitivity. J Pathol 226, 666-673 .10.1002/path.3026
[23] Suzuki, K., and Takahashi, K. (2008). Regulation of lamellipodia formation and cell invasion by CLIP-170 in invasive human breast cancer cells. Biochem Biophys Res Commun 368, 199-204 .10.1016/j.bbrc.2008.01.069
[24] Vicente-Manzanares, M., and Horwitz, A.R. (2011). Cell migration: an overview. Methods Mol Biol 769, 1-24 .10.1007/978-1-61779-207-6_1
[25] Wang, H., Liu, B., Zhang, C., Peng, G., Liu, M., Li, D., Gu, F., Chen, Q., Dong, J.T., Fu, L., . (2009). Parkin regulates paclitaxel sensitivity in breast cancer via a microtubule-dependent mechanism. J Pathol 218, 76-85 .10.1002/path.2512
[26] Watanabe, T., Noritake, J., and Kaibuchi, K. (2005). Regulation of microtubules in cell migration. Trends Cell Biol 15, 76-83 .10.1016/j.tcb.2004.12.006
[27] Weis, S.M., and Cheresh, D.A. (2011). Tumor angiogenesis: molecular pathways and therapeutic targets. Nat Med 17, 1359-1370 .10.1038/nm.2537