Anti-angiogenic action of the C-terminal domain of tenomodulin that shares homology with chondromodulin-I.
Tenomodulin (TeM) is a type II transmembrane glycoprotein that contains a C-terminal domain with homology to the mature, secreted form of chondromodulin-I (ChM-I), a cartilage-derived angiogenesis inhibitor. TeM transcripts have been found in hypovascular tissues such as tendons and ligaments but the biological activity of TeM has not yet been fully explored. Using an adenovirus expression system, we utilized the forced expression and subsequent secretion of the human TeM C-terminal 116 amino acids (Ad-shTeM) in human umbilical vein endothelial cells (HUVECs) to assess the anti-angiogenic properties of TeM. The C-terminal 120 amino acids of the human ChM-I precursor (Ad-shChM-I) was similarly expressed in HUVECs as a comparison study. Transduction of both Ad-shTeM and Ad-shChM-I resulted in significant impairment of the tube-forming activity of HUVECs, when cultured in Matrigel. Similarly, conditioned medium from COS7 cells, transfected with plasmid DNA encoding shTeM or shChM-I, inhibited tube formation of HUVECs when compared to medium derived from either COS7 cells transfected with control vector or from non-transfected cells. Upon infection of HUVECs with Ad-shTeM or Ad-shChM-I, DNA synthesis stimulated by vascular endothelial growth factor (VEGF) was reduced to 40-50% of normal levels. Additionally, in a modified Boyden chamber assay, migration of HUVECs in response to VEGF was significantly affected following transduction of either Ad-shTeM or Ad-shChM-I and these transduced HUVECs were found to spread well on type I collagen or fibronectin, but not on vitronectin. Furthermore, the transduction of either Ad-shTeM or Ad-shChM-I in human melanoma cells resulted in suppression of tumor growth in association with decreased vessel density in vivo. Hence, we have demonstrated that, similarly to ChM-1, the C-terminal domain of TeM exhibits both anti-angiogenic and anti-tumor activities when expressed in a secreted form.
Oshima
Yusuke
Y
Department of Cellular Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.
Sato
Koji
K
Tashiro
Fumi
F
Miyazaki
Jun-ichi
J
Nishida
Koji
K
Hiraki
Yuji
Y
Tano
Yasuo
Y
Shukunami
Chisa
C
D000256:Adenoviridae / Q000235:genetics; D020533:Angiogenesis Inhibitors / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003094:Collagen / Q000378:metabolism; D024042:Collagen Type I / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis / Q000187:drug effects; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000235:genetics; D007797:Laminin / Q000378:metabolism; D008544:Melanocytes / Q000187:drug effects / Q000378:metabolism; D008545:Melanoma / Q000473:pathology; D008546:Melanoma, Experimental / Q000473:pathology; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D009368:Neoplasm Transplantation; D017434:Protein Structure, Tertiary; D011509:Proteoglycans / Q000378:metabolism; D014184:Transplantation, Homologous; D014407:Tumor Cells, Cultured; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology
D020533:Angiogenesis Inhibitors; C489408:Cnmd protein, mouse; D024042:Collagen Type I; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D005353:Fibronectins; D036341:Intercellular Signaling Peptides and Proteins; D007797:Laminin; D008565:Membrane Proteins; D011509:Proteoglycans; C422444:TNMD protein, human; C486026:Tnmd protein, mouse; D042461:Vascular Endothelial Growth Factor A; C059485:matrigel; C068675:CNMD protein, human; D003094:Collagen; D004247:DNA