Top

Published in:

01-02-2019 | Original Paper

The calcium-binding type III repeats domain of thrombospondin-2 binds to fibroblast growth factor 2 (FGF2)

Authors: Marco Rusnati, Patrizia Borsotti, Elisabetta Moroni, Chiara Foglieni, Paola Chiodelli, Laura Carminati, Denise Pinessi, Douglas S. Annis, Giulia Paiardi, Antonella Bugatti, Alessandro Gori, Renato Longhi, Dorina Belotti, Deane F. Mosher, Giorgio Colombo, Giulia Taraboletti

Published in: Angiogenesis | Issue 1/2019

Abstract

Thrombospondin (TSP)-1 and TSP-2 share similar structures and functions, including a remarkable antiangiogenic activity. We have previously demonstrated that a mechanism of the antiangiogenic activity of TSP-1 is the interaction of its type III repeats domain with fibroblast growth factor-2 (FGF2), affecting the growth factor bioavailability and angiogenic activity. Since the type III repeats domain is conserved in TSP-2, this study aimed at investigating whether also TSP-2 retained the ability to interact with FGF2. The FGF2 binding properties of TSP-1 and TSP-2 and their recombinant domains were analyzed by solid-phase binding and surface plasmon resonance assays. TSP-2 bound FGF2 with high affinity (Kd = 1.3 nM). TSP-2/FGF2 binding was inhibited by calcium and heparin. The FGF2-binding domain of TSP-2 was located in the type III repeats and the minimal interacting sequence was identified as the GVTDEKD peptide in repeat 3C, corresponding to KIPDDRD, the active sequence of TSP-1. A second putative FGF2 binding sequence was also identified in repeat 11C of both TSPs. Computational docking analysis predicted that both the TSP-2 and TSP-1-derived heptapeptides interacted with FGF2 with comparable binding properties. Accordingly, small molecules based on the TSP-1 active sequence blocked TSP-2/FGF2 interaction. Binding of TSP-2 to FGF2 impaired the growth factor ability to interact with its cellular receptors, since TSP-2-derived fragments prevented the binding of FGF2 to both heparin (used as a structural analog of heparan sulfate proteoglycans) and FGFR-1. These findings identify TSP-2 as a new FGF2 ligand that shares with TSP-1 the same molecular requirements for interaction with the growth factor and a comparable capacity to block FGF2 interaction with proangiogenic receptors. These features likely contribute to TSP-2 antiangiogenic and antineoplastic activity, providing the rationale for future therapeutic applications.

Available only for authorised users

Carlson CB, Lawler J, Mosher DF (2008) Structures of thrombospondins. Cell Mol Life Sci 65:672–686CrossRefPubMedPubMedCentral

Adams JC, Lawler J (2011) The thrombospondins. Cold Spring Harb Perspect Biol 3:a009712. https://doi.org/10.1101/cshperspect.a009712 CrossRefPubMedPubMedCentral

Armstrong LC, Bornstein P (2003) Thrombospondins 1 and 2 function as inhibitors of angiogenesis. Matrix Biol 22:63–71CrossRefPubMed

Lawler PR, Lawler J (2012) Molecular basis for the regulation of angiogenesis by thrombospondin-1 and -2. Cold Spring Harb Perspect Med 2:a006627. https://doi.org/10.1101/cshperspect.a006627 CrossRefPubMedPubMedCentral

Volpert OV, Tolsma SS, Pellerin S et al (1995) Inhibition of angiogenesis by thrombospondin-2. Biochem Biophys Res Commun 217:326–332. https://doi.org/10.1006/bbrc.1995.2780 CrossRefPubMed

Panetti TS, Chen H, Misenheimer TM et al (1997) Endothelial cell mitogenesis induced by LPA: inhibition by thrombospondin-1 and thrombospondin-2. J Lab Clin Med 129:208–216CrossRefPubMed

Armstrong LC, Björkblom B, Hankenson KD et al (2002) Thrombospondin 2 inhibits microvascular endothelial cell proliferation by a caspase-independent mechanism. Mol Biol Cell 13:1893–1905CrossRefPubMedPubMedCentral

Kyriakides TR, Zhu YH, Smith LT et al (1998) Mice that lack thrombospondin 2 display connective tissue abnormalities that are associated with disordered collagen fibrillogenesis, an increased vascular density, and a bleeding diathesis. J Cell Biol 140:419–430CrossRefPubMedPubMedCentral

Kyriakides TR, Tam JW, Bornstein P (1999) Accelerated wound healing in mice with a disruption of the thrombospondin 2 gene. J Invest Dermatol 113:782–787. https://doi.org/10.1046/j.1523-1747.1999.00755.x CrossRefPubMed

10.

Hawighorst T, Velasco P, Streit M et al (2001) Thrombospondin-2 plays a protective role in multistep carcinogenesis: a novel host anti-tumor defense mechanism. EMBO J 20:2631–2640. https://doi.org/10.1093/emboj/20.11.2631 CrossRefPubMedPubMedCentral

11.

Kunstfeld R, Hawighorst T, Streit M et al (2014) Thrombospondin-2 overexpression in the skin of transgenic mice reduces the susceptibility to chemically induced multistep skin carcinogenesis. J Dermatol Sci 74:106–115. https://doi.org/10.1016/j.jdermsci.2014.01.002 CrossRefPubMedPubMedCentral

12.

Sun R, Wu J, Chen Y et al (2014) Down regulation of Thrombospondin2 predicts poor prognosis in patients with gastric cancer. Mol Cancer 13:225. https://doi.org/10.1186/1476-4598-13-225 CrossRefPubMedPubMedCentral

13.

Tokunaga T, Nakamura M, Oshika Y et al (1999) Thrombospondin 2 expression is correlated with inhibition of angiogenesis and metastasis of colon cancer. Br J Cancer 79:354–359. https://doi.org/10.1038/sj.bjc.6690056 CrossRefPubMedPubMedCentral

14.

Oshika Y, Masuda K, Tokunaga T et al (1998) Thrombospondin 2 gene expression is correlated with decreased vascularity in non-small cell lung cancer. Clin Cancer Res 4:1785–1788PubMed

15.

Dvorkina M, Nieddu V, Chakelam S et al (2016) A Promyelocytic leukemia protein-thrombospondin-2 axis and the risk of relapse in neuroblastoma. Clin Cancer Res 22:3398–3409. https://doi.org/10.1158/1078-0432.CCR-15-2081 CrossRefPubMed

16.

Chijiwa T, Abe Y, Ikoma N et al (2009) Thrombospondin 2 inhibits metastasis of human malignant melanoma through microenvironment-modification in NOD/SCID/gammaCnull (NOG) mice. Int J Oncol 34:5–13PubMed

17.

Streit M, Riccardi L, Velasco P et al (1999) Thrombospondin-2: a potent endogenous inhibitor of tumor growth and angiogenesis. Proc Natl Acad Sci USA 96:14888–14893CrossRefPubMed

18.

Simantov R, Febbraio M, Silverstein RL (2005) The antiangiogenic effect of thrombospondin-2 is mediated by CD36 and modulated by histidine-rich glycoprotein. Matrix Biol 24:27–34. https://doi.org/10.1016/j.matbio.2004.11.005 CrossRefPubMed

19.

Isenberg JS, Annis DS, Pendrak ML et al (2009) Differential interactions of thrombospondin-1, -2, and -4 with CD47 and effects on cGMP signaling and ischemic injury responses. J Biol Chem 284:1116–1125. https://doi.org/10.1074/jbc.M804860200 CrossRefPubMedPubMedCentral

20.

Calabro NE, Kristofik NJ, Kyriakides TR (2014) Thrombospondin-2 and extracellular matrix assembly. Biochim Biophys Acta 1840:2396–2402. https://doi.org/10.1016/j.bbagen.2014.01.013 CrossRefPubMedPubMedCentral

21.

Resovi A, Pinessi D, Chiorino G, Taraboletti G (2014) Current understanding of the thrombospondin-1 interactome. Matrix Biol 37:83–91. https://doi.org/10.1016/j.matbio.2014.01.012 CrossRefPubMed

22.

Margosio B, Marchetti D, Vergani V et al (2003) Thrombospondin 1 as a scavenger for matrix-associated fibroblast growth factor 2. Blood 102:4399–4406CrossRefPubMed

23.

Margosio B, Rusnati M, Bonezzi K et al (2008) Fibroblast growth factor-2 binding to the thrombospondin-1 type III repeats, a novel antiangiogenic domain. Int J Biochem Cell Biol 40:700–709CrossRefPubMed

24.

Taraboletti G, Belotti D, Borsotti P et al (1997) The 140-kilodalton antiangiogenic fragment of thrombospondin-1 binds to basic fibroblast growth factor. Cell Growth Differ 8:471–479PubMed

25.

Colombo G, Margosio B, Ragona L et al (2010) Non-peptidic thrombospondin-1 mimics as fibroblast growth factor-2 inhibitors: an integrated strategy for the development of new antiangiogenic compounds. J Biol Chem 285:8733–8742. https://doi.org/10.1074/jbc.M109.085605 CrossRefPubMedPubMedCentral

26.

Annis DS, Murphy-Ullrich JE, Mosher DF (2006) Function-blocking antithrombospondin-1 monoclonal antibodies. J Thromb Haemost 4:459–468CrossRefPubMedPubMedCentral

27.

Mosher DF, Huwiler KG, Misenheimer TM, Annis DS (2002) Expression of recombinant matrix components using baculoviruses. Methods Cell Biol 69:69–81CrossRefPubMed

28.

Amblard M, Fehrentz J-A, Martinez J, Subra G (2006) Methods and protocols of modern solid phase peptide synthesis. Mol Biotechnol 33:239–254. https://doi.org/10.1385/MB:33:3:239 CrossRefPubMed

29.

Foglieni C, Pagano K, Lessi M et al (2016) Integrating computational and chemical biology tools in the discovery of antiangiogenic small molecule ligands of FGF2 derived from endogenous inhibitors. Sci Rep 6:23432. https://doi.org/10.1038/srep23432 CrossRefPubMedPubMedCentral

30.

Pagano K, Torella R, Foglieni C et al (2012) Direct and allosteric inhibition of the FGF2/HSPGs/FGFR1 ternary complex formation by an antiangiogenic, thrombospondin-1-mimic small molecule. PLoS ONE 7:e36990. https://doi.org/10.1371/journal.pone.0036990 CrossRefPubMedPubMedCentral

31.

Misenheimer TM, Mosher DF (2005) Biophysical characterization of the signature domains of thrombospondin-4 and thrombospondin-2. J Biol Chem 280:41229–41235. https://doi.org/10.1074/jbc.M504696200 CrossRefPubMedPubMedCentral

32.

Misenheimer TM, Hannah BL, Annis DS, Mosher DF (2003) Interactions among the three structural motifs of the C-terminal region of human thrombospondin-2. Biochemistry 42:5125–5132CrossRefPubMed

33.

Carlson CB, Bernstein DA, Annis DS et al (2005) Structure of the calcium-rich signature domain of human thrombospondin-2. Nat Struct Mol Biol 12:910–914CrossRefPubMedPubMedCentral

34.

Kvansakul M, Adams JC, Hohenester E (2004) Structure of a thrombospondin C-terminal fragment reveals a novel calcium core in the type 3 repeats. Embo J 23:1223–1233CrossRefPubMedPubMedCentral

35.

Hannah BL, Misenheimer TM, Annis DS, Mosher DF (2003) A polymorphism in thrombospondin-1 associated with familial premature coronary heart disease causes a local change in conformation of the Ca²⁺-binding repeats. J Biol Chem 278:8929–8934CrossRefPubMed

36.

Hannah BL, Misenheimer TM, Pranghofer MM, Mosher DF (2004) A polymorphism in thrombospondin-1 associated with familial premature coronary artery disease alters Ca²⁺ binding. J Biol Chem 279:51915–51922CrossRefPubMed

37.

Giacomini A, Chiodelli P, Matarazzo S et al (2016) Blocking the FGF/FGFR system as a “two-compartment” antiangiogenic/antitumor approach in cancer therapy. Pharmacol Res 107:172–185. https://doi.org/10.1016/j.phrs.2016.03.024 CrossRefPubMed

38.

Chiodelli P, Bugatti A, Urbinati C, Rusnati M (2015) Heparin/Heparan sulfate proteoglycans glycomic interactome in angiogenesis: biological implications and therapeutical use. Molecules 20:6342–6388. https://doi.org/10.3390/molecules20046342 CrossRefPubMedPubMedCentral

39.

Rusnati M, Presta M (2015) Angiogenic growth factors interactome and drug discovery: the contribution of surface plasmon resonance. Cytokine Growth Factor Rev 26:293–310. https://doi.org/10.1016/j.cytogfr.2014.11.007 CrossRefPubMed

40.

Rusnati M, Bugatti A, Mitola S et al (2009) Exploiting surface plasmon resonance (SPR) technology for the identification of fibroblast growth factor-2 (FGF2) antagonists endowed with antiangiogenic activity. Sensors (Basel) 9:6471–6503. https://doi.org/10.3390/s90806471 CrossRef

41.

Muppala S, Frolova E, Xiao R et al (2015) Proangiogenic properties of thrombospondin-4. Arterioscler Thromb Vasc Biol 35:1975–1986. https://doi.org/10.1161/ATVBAHA.115.305912 CrossRefPubMedPubMedCentral

42.

Annis DS, Gunderson KA, Mosher DF (2007) Immunochemical analysis of the structure of the signature domains of thrombospondin-1 and thrombospondin-2 in low calcium concentrations. J Biol Chem 282:27067–27075. https://doi.org/10.1074/jbc.M703804200 CrossRefPubMed

43.

Turner N, Grose R (2010) Fibroblast growth factor signalling: from development to cancer. Nat Rev Cancer 10:116–129. https://doi.org/10.1038/nrc2780 CrossRefPubMed

44.

Wesche J, Haglund K, Haugsten EM (2011) Fibroblast growth factors and their receptors in cancer. Biochem J 437:199–213. https://doi.org/10.1042/BJ20101603 CrossRef

45.

Stenina-Adognravi O (2014) Invoking the power of thrombospondins: regulation of thrombospondins expression. Matrix Biol 37:69–82. https://doi.org/10.1016/j.matbio.2014.02.001 CrossRefPubMed

46.

Tooney PA, Sakai T, Sakai K et al (1998) Restricted localization of thrombospondin-2 protein during mouse embryogenesis: a comparison to thrombospondin-1. Matrix Biol 17:131–143CrossRefPubMed

47.

Kyriakides TR, Maclauchlan S (2009) The role of thrombospondins in wound healing, ischemia, and the foreign body reaction. J Cell Commun Signal 3:215–225. https://doi.org/10.1007/s12079-009-0077-z CrossRefPubMedPubMedCentral

48.

Agah A, Kyriakides TR, Lawler J, Bornstein P (2002) The lack of thrombospondin-1 (TSP1) dictates the course of wound healing in double-TSP1/TSP2-null mice. Am J Pathol 161:831–839. https://doi.org/10.1016/S0002-9440(10)64243-5 CrossRefPubMedPubMedCentral

49.

Murphy-Ullrich JE, Suto MJ (2017) Thrombospondin-1 regulation of latent TGF-β activation: a therapeutic target for fibrotic disease. Matrix Biol. https://doi.org/10.1016/j.matbio.2017.12.009 CrossRefPubMedPubMedCentral

50.

Schultz-Cherry S, Chen H, Mosher DF et al (1995) Regulation of transforming growth factor-b activation by discrete sequences of thrombospondin 1. J Biol Chem 270:7304–7310CrossRefPubMed

51.

Taraboletti G, Rusnati M, Ragona L, Colombo G (2010) Targeting tumor angiogenesis with TSP-1-based compounds: rational design of antiangiogenic mimetics of endogenous inhibitors. Oncotarget 1:662–673CrossRef

52.

Koch M, Hussein F, Woeste A et al (2011) CD36-mediated activation of endothelial cell apoptosis by an N-terminal recombinant fragment of thrombospondin-2 inhibits breast cancer growth and metastasis in vivo. Breast Cancer Res Treat 128:337–346. https://doi.org/10.1007/s10549-010-1085-7 CrossRefPubMed

Title: The calcium-binding type III repeats domain of thrombospondin-2 binds to fibroblast growth factor 2 (FGF2)
Authors: Marco Rusnati
Patrizia Borsotti
Elisabetta Moroni
Chiara Foglieni
Paola Chiodelli
Laura Carminati
Denise Pinessi
Douglas S. Annis
Giulia Paiardi
Antonella Bugatti
Alessandro Gori
Renato Longhi
Dorina Belotti
Deane F. Mosher
Giorgio Colombo
Giulia Taraboletti
Publication date: 01-02-2019
Publisher: Springer Netherlands
Published in: Angiogenesis / Issue 1/2019
Print ISSN: 0969-6970
Electronic ISSN: 1573-7209
DOI: https://doi.org/10.1007/s10456-018-9644-3

ACC 2024 Congress Coverage

Heart Failure Video

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

The calcium-binding type III repeats domain of thrombospondin-2 binds to fibroblast growth factor 2 (FGF2)

Abstract

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 1/2019

Inhibition of macrophage inflammatory protein-1β improves endothelial progenitor cell function and ischemia-induced angiogenesis in diabetes

Extracellular vesicles of multiple myeloma cells utilize the proteasome inhibitor mechanism to moderate endothelial angiogenesis

Perfused 3D angiogenic sprouting in a high-throughput in vitro platform

Angiogenesis in pancreatic cancer: current research status and clinical implications

Pazopanib may reduce bleeding in hereditary hemorrhagic telangiectasia

CMTM4 regulates angiogenesis by promoting cell surface recycling of VE-cadherin to endothelial adherens junctions

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page