Abstract
Thrombospondin-1 (TSP-1) is a naturally occurring inhibitor of angiogenesis that limits vessel density in normal tissues and curtails tumor growth. Here, we show that the inhibition of angiogenesis in vitro and in vivo and the induction of apoptosis by thrombospondin-1 all required the sequential activation of CD36, p59fyn, caspase-3 like proteases and p38 mitogen-activated protein kinases. We also detected increased endothelial cell apoptosis in situ at the margins of tumors in mice treated with thrombospondin-1. These results indicate that thrombospondin-1, and possibly other broad-spectrum natural inhibitors of angiogenesis, act in vivo by inducing receptor-mediated apoptosis in activated microvascular endothelial cells.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Hanahan, D. & Folkman, J. Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell 86, 353–364 (1996).
Bouck, N., Stellmach, V. & Hsu, S. How tumors become angiogenic. Adv. Cancer Res. 69, 135–174 ( 1996).
Volpert, O.V., Lawler, J. & Bouck, N.P. A human fibrosarcoma inhibits systemic angiogenesis and the growth of experimental metastases via thrombospondin-1. Proc. Natl. Acad. Sci. USA 95, 6343–6348 (1998)
O'Reilly, M.S. et al. Angiostatin: a novel angiogenesis inhibitor that mediates the suppression of metastases by a Lewis lung carcinoma. Cell 79, 315–328 (1994).
O'Reilly, M.S. et al. Endostatin: an endogenous inhibitor of angiogenesis and tumor growth. Cell 88, 277–285 (1997).
Boehm, T., Folkman, J., Browder, T. & O'Reilly, M.S. Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance. Nature 390, 404–407 ( 1997).
Guo, N., Krutzsch, H.C., Inman, J.K. & Roberts, D.D. Thrombospondin 1 and type I repeat peptides of thrombospondin 1 specifically induce apoptosis of endothelial cells. Cancer Res. 57, 1735–1742 (1997).
Claesson-Welsh, L. et al. Angiostatin induces endothelial cell apoptosis and activation of focal adhesion kinase independently of the integrin-binding motif RGD. Proc. Natl. Acad. Sci. USA 95, 5579– 5583 (1998).
Lucas, R. et al. Multiple forms of angiostatin induce apoptosis in endothelial cells. Blood 92, 4730–4741 (1998).
Dhanabal, M., et al. Endostatin induces endothelial cell apoptosis. J. Biol. Chem. 274, 11721–11726 (1999).
Yue, T.L. et al. 2-Methoxyestradiol, an endogenous estrogen metabolite, induces apoptosis in endothelial cells and inhibits angiogenesis: possible role for stress-activated protein kinase signaling pathway and Fas expression. Mol. Pharmacol. 51, 951–962 (1992).
DiPietro, L.A. in Regulation of Angiogenesis (eds. Goldberg. I.D. & Rosen, E.M.) 295–313 (Birkhauser Verlag, Basel, Switzerland, 1997).
Roberts, D.D. Regulation of tumor growth and metastasis by thrombospondin-1. FASEB J. 10, 1183–1191 ( 1996).
Dawson, D.W. & Bouck, N. in Antiangiogenic Agents in Cancer (ed. Teicher, B.A.) 185–204 (Humana, Totowa, New Jersey, 1998).
Crawford, S.E. et al. Thrombospondin-1 is a major activator of TGF-β1 in vivo. Cell 93, 1159– 1170 (1998).
Weinstat-Saslow, D.L. et al. Transfection of thrombospondin 1 complementary DNA into a human breast carcinoma cell line reduces primary tumor growth, metastatic potential, and angiogenesis. Cancer Res. 54, 6504–6511 (1994).
Castle, V.P., Dixit, V.M. & Polverini, P.J. Thrombospondin-1 suppresses tumorigenesis and angiogenesis in serum-and anchorage-independent NIH 3T3 cells. Lab. Invest. 77, 51–61 (1997).
Bleuel, K., Popp, S., Fusenig, N.E., Stanbridge, E.J. & Boukamp, P. Tumor suppression in human skin carcinoma cells by chromosome 15 transfer or thrombospondin-1 overexpression through halted tumor vascularization . Proc. Natl. Acad. Sci. USA 96, 2065– 2070 (1999).
Volpert, O.V. et al. Inhibition of angiogenesis by thrombospondin-2. Biochem. Biophys. Res. Comm. 217, 326– 332 (1995).
Dawson, D.W. et al. CD36 mediates the in vitro inhibitory effects of thrombospondin-1 on endothelial cells. J. Cell Biol. 138, 707–717 (1997).
Huang, M.M., Bolen, J.B., Barnwell, J.W., Shattil, S.J. & Brugge, J.S. Membrane glycoprotein IV (CD36) is physically associated with the Fyn, Lyn, and Yes protein-tyrosine kinases in human platelets. Proc. Natl. Acad. Sci. USA 88, 7844–7848 (1991).
Bull, H.A., Brickell, P.M. & Dowd, P.M. Src-related protein tyrosine kinases are physically associated with the surface antigen CD36 in human dermal microvascular endothelial cells. FEBS Lett. 351, 41– 44 (1994).
Tolsma, S.S. et al. Peptides derived from two separate domains of the matrix protein thrombospondin-1 have anti-angiogenic activity. J. Cell Biol. 122, 497–511 (1993).
Cosulich, S. & Clarke, P. Apoptosis: does stress kill?. Curr. Biol. 6, 1586–1588 (1996).
Le-Niculescu, H. et al. Withdrawal of survival factors results in activation of the JNK pathway in neuronal cells leading to Fas ligand induction and cell death . Mol. Cell. Biol. 19, 751– 763 (1999).
Verheij, M. et al. Requirement for ceramide-initiated SAPK/JNK signaling in stress-induced apoptosis. Nature 380, 75– 79 (1996).
Ichijo, H. et al. Induction of apoptosis by ASK1, a mammalian MAPKKK that activates SAPK/JNK and p38 signaling pathways. Science 275, 90–94 (1997).
Kumar, S. et al. Activation of the HIV-1 long terminal repeat by cytokines and environmental stress requires an active CSBP/p38 MAP kinase. J. Biol. Chem. 271, 30864–30869 (1996).
Polverini, P.J., Bouck, N.P. & Rastinejad, F. Assay and purification of naturally occurring inhibitor of angiogenesis. Meth. Enzymol. 198, 440 –450 (1991).
Schultz-Cherry, S. & Murphy-Ullrich, J.E. Thrombospondin causes activation of latent transforming growth factor-beta secreted by endothelial cells by a novel mechanism. J. Cell Biol. 122, 923–932 (1993).
Storgard, C.M. et al. Decreased angiogenesis and arthritic disease in rabbits treated with an αvβ3 antagonist. J. Clin. Invest. 103, 47–54 (1999).
Jacobson, M.D., Weil, M. & Raff, M.C. Programmed cell death in animal development. Cell 88, 347–354 ( 1997).
Dragovich, T., Rudin, C.M. & Thompson, C.B. Signal transduction pathways that regulate cell survival and cell death. Oncogene 17, 3207– 3213 (1998).
Nor, J.E., Christensen, J., Mooney, D.J. & Polverini, P.J. Vascular endothelial growth factor (VEGF)-mediated angiogenesis is associated with enhanced endothelial cell survival and induction of Bcl-2 expression . Am J. Pathol. 154, 375– 384 (1999).
Benjamin, L.E., Golijanin, D., Itin, A., Pode, D. & Keshet, E. Selective ablation of immature blood vessels in established human tumors follows vascular endothelial growth factor withdrawal. J. Clin. Invest. 103, 159–165 (1999).
Gupta, K. et al. VEGF prevents apoptosis of human microvascular endothelial cells via opposing effects on MAPK/ERK and SAPK/JNK signaling. Exp. Cell Res. 247, 495–504 ( 1999).
Erpel, T. & Courtneidge, S.A. Src family protein tyrosine kinases and cellular signal transduction pathways. Curr. Opin. Cell Biol. 7, 176–182 ( 1995).
Miyakawa, T. et al. Fyn-kinase as a determinant of ethanol sensitivity: relation to NMDA-receptor function. Science 278, 698–701 (1997).
Grant, S.G.N. et al. Impaired long-term potentiation, spatial learning, and hippocampal development in fyn mutant mice. Science 258, 1903–1910 (1992).
Juo, P., et al. Fas activation of the p38 mitogen-activated protein kinase signalling pathway requires ICE/CED-3 family proteases. Mol. Cell. Biol. 17, 24–35 (1997).
Cardone, M.H., Salvesen, G.S., Widmann, C., Johnson, G. & Frisch, S.M. The regulation of anoikis: MEKK-1 activation requires cleavage by caspases. Cell 90, 315–323 (1997).
Kothakota, S. et al. Caspase-3-generated fragment of gelsolin: effector of morphological change in apoptosis. Science 278, 294– 298 (1997).
Salazar, R., Bell, S.E. & Davis, G.E. Coordinate induction of the actin cytoskeletal regulatory proteins gelsolin, vasodilator-stimulated phosphoprotein, and profilin during capillary morphogenesis in vitro. Exp. Cell Res. 249, 22–32 (1999).
Brooks, P.C. et al. Integrin alphaV beta3 antagonists promote tumor regression by inducing apoptosis of angiogenic blood vessels. Cell 79, 1157–1164 (1994).
Stromblad, S., Becker, J.C., Yebra, M., Brooks, P.C. & Cheresh, D.A. Suppression of p53 activity and p21WAF1/CIP1 expression by vascular cell integrin alphaVbeta3 during angiogenesis. J. Clin. Invest. 98, 426–433 ( 1996).
Streilein, J.W. Unraveling immune privilege. Science 270, 1158–1159 (1995).
Schlaepfer, D.D. & Hunter, T. Evidence for in vivo phosphorylation of the Grb2 SH2-domain binding site on focal adhesion kinase by Src-family protein-tyrosine kinases. Mol. Cell. Biol. 16, 5623–5633 ( 1996).
Febbraio, M., Abumrad, N.A., Haijar, D.P., Sharma, K., Pearce, S.F.A. & Silverstein, R.L. A null mutation in murine CD36 reveals an important role in fatty acid and lipoprotein metabolism. J. Biol. Chem. 274, 19055–19062 (1999).
Acknowledgements
We thank G. Soff (Northwestern University) and M. O'Reilly (Harvard University), for supplying active angiostatin, X. Huang and L. Huang for technical assistance, and the National Cancer Institute for support through grants CA52750 and CA64239. B.J. acknowledges support from a NATO fellowship and also thanks F. Dolfi, J. M. Redondo and P. Gómez del Arco for their advice on kinase assays, D. Dawson for discussions, and A. Munoz for his support.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Jiménez, B., Volpert, O., Crawford, S. et al. Signals leading to apoptosis-dependent inhibition of neovascularization by thrombospondin-1. Nat Med 6, 41–48 (2000). https://doi.org/10.1038/71517
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/71517
This article is cited by
-
Thrombospondin 1 enhances systemic inflammation and disease severity in acute-on-chronic liver failure
BMC Medicine (2024)
-
Disclosing the molecular profile of the human amniotic mesenchymal stromal cell secretome by filter-aided sample preparation proteomic characterization
Stem Cell Research & Therapy (2023)
-
Alleviating hypoxia to improve cancer immunotherapy
Oncogene (2023)
-
Exosomes derived from stem cells of human deciduous exfoliated teeth inhibit angiogenesis in vivo and in vitro via the transfer of miR-100-5p and miR-1246
Stem Cell Research & Therapy (2022)
-
METTL14 promotes prostate tumorigenesis by inhibiting THBS1 via an m6A-YTHDF2-dependent mechanism
Cell Death Discovery (2022)