Antagonism of VEGF-A-induced increase in vascular permeability by an integrin α3β1-Shp-1-cAMP/PKA pathway.
In cancer, VEGF-induced increase in vascular permeability results in increased interstitial pressure, reducing perfusion and increasing hypoxia, which reduce delivery of chemotherapeutic agents and increase resistance to ionizing radiation. Here, we show that both TIMP-2 and Ala + TIMP-2, a TIMP-2 mutant without matrix metalloproteinase inhibitory activity, antagonize the VEGF-A-induced increase in vascular permeability, both in vitro and in vivo. Like other agents known to preserve endothelial barrier function, TIMP-2 elevates cytosolic levels of cAMP and increases cytoskeletal-associated vascular endothelial cadherin in human microvascular endothelial cells. All of these effects are completely ablated by selective knockdown of integrin α3β1 expression, expression of a dominant negative protein tyrosine phosphatase Shp-1 mutant, administration of the protein tyrosine phosphatase inhibitor orthovanadate, or the adenylate cyclase inhibitor SQ22536. This TIMP-2-mediated inhibition of vascular permeability involves an integrin α3β1-Shp-1-cAMP/protein kinase A-dependent vascular endothelial cadherin cytoskeletal association, as evidenced by using siRNAs to integrin α3β1 and Shp-1, or treatment with Shp-1 inhibitor NSC87877 and protein kinase A inhibitor H89. Our results demonstrate the potential utility for TIMP-2 in cancer therapy through "normalization" of vascular permeability in addition to previously described antiangiogenic effects.
Department of Molecular Bioscience, College of Biomedical Science, Kangwon National University, Chuncheon, Korea.
Soo Hyeon
SH
Kim
Young-Rak
YR
Cho
Hyeon-Ju
HJ
Kim
Joa Sub
JS
Oh
Eun-Kyung
EK
Ahn
Hye-Jin
HJ
Ko
Byung Joon
BJ
Hwang
Seo-Jin
SJ
Lee
Yongwan
Y
Cho
Yong Kee
YK
Kim
William G
WG
Stetler-Stevenson
Dong-Wan
DW
Seo
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D002199:Capillary Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004336:Drug Antagonism; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D039201:Integrin alpha3beta1 / Q000235:genetics / Q000378:metabolism; D007546:Isoquinolines / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence; D009154:Mutation; D011485:Protein Binding / Q000187:drug effects; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D015398:Signal Transduction / Q000187:drug effects*; D013449:Sulfonamides / Q000494:pharmacology; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics / Q000494:pharmacology*; D014638:Vanadates / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*; D000099152:Cadherin 5
D000225:Adenine; D015703:Antigens, CD; D015820:Cadherins; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D039201:Integrin alpha3beta1; D007546:Isoquinolines; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D013449:Sulfonamides; D019716:Tissue Inhibitor of Metalloproteinase-2; D014638:Vanadates; D042461:Vascular Endothelial Growth Factor A; D000099152:Cadherin 5; C017759:9-(tetrahydro-2-furyl)-adenine; C509249:PTPN6 protein, human; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide