Lumen formation and other angiogenic activities of cultured capillary endothelial cells are inhibited by thrombospondin-1.
The large secreted glycoprotein thrombospondin-1 is a potent inhibitor of neovascularization in vivo. In order to better understand its mechanism of action, we have determined the full range of deficits thrombospondin can impose on cultured capillary endothelial cells. Exogenously added thrombospondin-1 blocked the ability of these cells to organize into cords. It blocked the migration of endothelial cells and vascular smooth muscle cells, but not that of fibroblasts, neutrophils, or keratinocytes, demonstrating specificity. Conversely, when the endogenous thrombospondin-1 produced by the endothelial cells was inactivated using antibodies that can neutralize its inhibition of neovascularization in vivo, migration toward basic fibroblast growth factor and cord formation were stimulated, and sparsely plated cells developed cylindrical cavities. These cavities formed by vesicle fusion, extended the depth of the cell, and appeared to be incipient lumens, staining positively for the luminal marker angiotensin converting enzyme. Antiangiogenic levels of thrombospondin-1 had no measurable effect on the overall level of activity of soluble gelatinases or on urokinase plasminogen activator produced by activated endothelial cells. Coupled with previously published data, these results demonstrate thrombospondin-1 is a multifaceted inhibitor able to block the entire program of dedifferentiation and redifferentiation essential to the formation of new vessels. They also support the contention that the endogenously produced protein contributes to the quiescence of the normal vasculature.
Tolsma
S S
SS
Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, Illinois, 60611, USA.
Stack
M S
MS
Bouck
N
N
D000818:Animals; D002196:Capillaries / Q000166:cytology*; D002417:Cattle; D002449:Cell Aggregation; D002461:Cell Line, Transformed; D002465:Cell Movement; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000737:chemistry; D003864:Depression, Chemical; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects; D018093:Gelatinases / Q000032:analysis; D015603:Keratinocytes / Q000187:drug effects; D007962:Leukocytes / Q000187:drug effects; D008562:Membrane Glycoproteins / Q000494:pharmacology*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D019699:Thrombospondins; D014568:Urokinase-Type Plasminogen Activator / Q000032:analysis
D017077:Culture Media, Conditioned; D008562:Membrane Glycoproteins; D019699:Thrombospondins; D016222:Fibroblast Growth Factor 2; D014568:Urokinase-Type Plasminogen Activator; D018093:Gelatinases