title: Thrombospondin-1 modulates VEGF signaling via CD36 by recruiting SHP-1 to VEGFR2 complex in microvascular endothelial cells.
abstract: Thrombospondin-1 (TSP-1) inhibits growth factor signaling at the receptor level in microvascular endothelial cells (MVEC), and CD36 has been suggested to be involved in this inhibition, but the mechanisms are not known. We hypothesized that CD36-TSP-1 interaction recruits Src homology 2 domain-containing protein tyrosine phosphatase (SHP)-1 to the vascular endothelial growth factor receptor 2 (VEGFR2) signaling complex and attenuates vascular endothelial growth factor (VEGF) signaling. Western blots of anti-CD36 and anti-VEGFR2 immunoprecipitates from VEGF-treated MVEC showed that exposure of the cells to a recombinant protein containing the CD36 binding domain of thrombospondin-1 (known as the TSR domain) induced association of SHP-1 with the VEGFR2/CD36 signaling complex and thereby suppressed VEGFR2 phosphorylation. SHP-1 phosphatase activity was increased in immunoprecipitated VEGFR2 complexes from TSR-treated cells. Silencing CD36 expression in MVEC by small interfering RNA (siRNA) or genetic deletion of cd36 in mice showed that TSR-induced SHP-1/VEGFR2 complex formation required CD36 in vitro and in vivo. Silencing SHP-1 expression in MVEC by siRNA abrogated TSR-mediated inhibition of VEGFR2 phosphorylation as well as TSR-mediated inhibition of VEGF-induced endothelial cell migration and tube formation. These studies reveal a SHP-1-mediated antiangiogenic pathway induced by CD36-TSP-1 interaction that inhibits VEGFR2 signaling and they provide a novel target to modulate angiogenesis therapeutically.
authors.affiliation: Department of Cell Biology, Lerner Research Institute, Cleveland Clinic, Cleveland OH, USA.
authors.forename: Ling-Yun
authors.initials: LY
authors.lastname: Chu
authors.forename: Devi Prasadh
authors.initials: DP
authors.lastname: Ramakrishnan
authors.forename: Roy L
authors.initials: RL
authors.lastname: Silverstein
mesh_terms: D000818:Animals; D018955:CD36 Antigens / Q000172:deficiency / Q000378:metabolism*; D002465:Cell Movement; D042783:Endothelial Cells / Q000378:metabolism*; D017353:Gene Deletion; D051379:Mice; D008810:Mice, Inbred C57BL; D055806:Microvessels / Q000166:cytology; D008954:Models, Biological; D018919:Neovascularization, Physiologic; D010766:Phosphorylation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6 / Q000378:metabolism*; D015398:Signal Transduction; D019700:Thrombospondin 1 / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism*
chemical_list: D018955:CD36 Antigens; D019700:Thrombospondin 1; D042461:Vascular Endothelial Growth Factor A; C467480:vascular endothelial growth factor A, mouse; D040301:Vascular Endothelial Growth Factor Receptor-2; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6
references.citation: J Mol Signal. 2008 Mar 31;3:8
references.pmid: 18377662
references.citation: FASEB J. 2009 Oct;23(10):3368-76
references.pmid: 19528255
references.citation: Nat Rev Mol Cell Biol. 2006 May;7(5):359-71
references.pmid: 16633338
references.citation: Biochim Biophys Acta. 1977 Feb 4;464(3):493-508
references.pmid: 836823
references.citation: Semin Thromb Hemost. 1987 Jul;13(3):298-306
references.pmid: 3317840
references.citation: Nature. 2005 Feb 3;433(7025):523-7
references.pmid: 15690042
references.citation: Biochem Biophys Res Commun. 2009 Apr 17;381(4):491-5
references.pmid: 19233143
references.citation: J Biol Chem. 1997 Dec 19;272(51):32521-7
references.pmid: 9405464
references.citation: J Biol Chem. 2006 Oct 13;281(41):31002-11
references.pmid: 16880211
references.citation: Biochem Biophys Res Commun. 1997 Nov 26;240(3):635-9
references.pmid: 9398617
references.citation: Biochem J. 2004 Mar 1;378(Pt 2):281-92
references.pmid: 14662007
references.citation: J Biol Chem. 2003 Feb 14;278(7):4668-74
references.pmid: 12468540
references.citation: J Clin Invest. 1987 Apr;79(4):1054-61
references.pmid: 2435757
references.citation: Trends Biochem Sci. 2005 Aug;30(8):430-6
references.pmid: 15996869
references.citation: PLoS One. 2012;7(7):e40033
references.pmid: 22808089
references.citation: J Biol Chem. 2001 Feb 2;276(5):3222-30
references.pmid: 11058584
references.citation: Blood. 2011 Apr 28;117(17):4658-66
references.pmid: 21378271
references.citation: EXS. 1997;79:295-314
references.pmid: 9002224
references.citation: Trends Biochem Sci. 2003 Jun;28(6):284-93
references.pmid: 12826400
references.citation: Sci Signal. 2009 May 26;2(72):re3
references.pmid: 19471024
references.citation: FASEB J. 2005 Oct;19(12):1713-5
references.pmid: 16037098
references.citation: Blood. 2011 Jun 2;117(22):6036-45
references.pmid: 21441463
references.citation: Mol Pharmacol. 2006 Aug;70(2):562-70
references.pmid: 16717135
references.citation: Proc Natl Acad Sci U S A. 1990 Sep;87(17):6624-8
references.pmid: 1697685
references.citation: J Biol Chem. 1992 Aug 15;267(23):16607-12
references.pmid: 1379600
references.citation: Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7844-8
references.pmid: 1715582
references.citation: Proc Natl Acad Sci U S A. 1997 Sep 2;94(18):9797-801
references.pmid: 9275205
references.citation: J Cell Sci. 2009 Sep 15;122(Pt 18):3385-92
references.pmid: 19706677
references.citation: Front Biosci (Landmark Ed). 2013 Jun 01;18:852-61
references.pmid: 23747852
references.citation: J Cell Biol. 1997 Aug 11;138(3):707-17
references.pmid: 9245797
references.citation: Stem Cells. 2010 Sep;28(9):1476-86
references.pmid: 20641039
references.citation: Nat Med. 2000 Jan;6(1):41-8
references.pmid: 10613822
references.citation: Blood. 2011 May 26;117(21):5744-50
references.pmid: 21427288
references.citation: J Clin Invest. 2009 Jan;119(1):136-45
references.pmid: 19065049
references.citation: Cell Metab. 2006 Sep;4(3):211-21
references.pmid: 16950138
references.citation: J Biol Chem. 2000 Nov 10;275(45):35264-75
references.pmid: 10956645
references.citation: J Biol Chem. 1999 Jul 2;274(27):19055-62
references.pmid: 10383407
references.citation: Atherosclerosis. 2007 Mar;191(1):33-9
references.pmid: 16725144
references.citation: Am J Physiol Heart Circ Physiol. 2009 May;296(5):H1344-51
references.pmid: 19304944
references.citation: Science. 1996 May 24;272(5265):1173-6
references.pmid: 8638162
references.citation: EMBO J. 2001 Jun 1;20(11):2768-78
references.pmid: 11387210
references.citation: Blood. 2001 Mar 15;97(6):1689-96
references.pmid: 11238109