Recombinant semaphorin 6A-1 ectodomain inhibits in vivo growth factor and tumor cell line-induced angiogenesis.
The Semaphorins are a large family of transmembrane, GPI-anchored and secreted proteins that play an important role in neuronal and endothelial cell guidance. A human gene related to the class 6 Semaphorin family, Semaphorin 6A-1 (Sema 6A-1) was identified by homology-based genomic mining. Recent implication of Sema 3 family members in tumor angiogenesis and our expression analysis of Sema 6A-1 suggested that class 6 Semaphorin might effect tumor neovascularization. The mRNA expression of Sema 6A-1 was elevated in several renal tumor tissue samples relative to adjacent nontumor tissue samples from the same patient. Sema 6A-1 transcript was also expressed in the majority of renal clear cell carcinoma (RCC) cell lines and to a lesser extent in endothelial cells. To test the role of Sema 6A-1 in tumor angiogenesis, we engineered, expressed and purified the Sema 6A-1 soluble extracellular domain (Sema-ECD). The purified Sema-ECD was screened in a variety of endothelial cell-based assays both in vitro and in vivo. In vitro, Sema-ECD blocked VEGF-mediated endothelial cell migration. These effects were explained in part by our observation in endothelial cells that Sema-ECD inhibited VEGF-mediated Src, FAK and ERK phosphorylation. In vivo, mouse Matrigel assays demonstrated that the intraperitoneal administration of recombinant Sema-ECD inhibited both bFGF/VEGF and tumor cell line-induced neovascularization. These findings reveal a novel therapeutic utility for Sema 6A-1 (Sema-ECD) as an inhibitor of growth factor as well as tumor-induced angiogenesis.
Dhanabal
Mohanraj
M
CuraGen Corporation, Branford, Connecticut, USA. mdhanabal@curagen.com
Wu
Frank
F
Alvarez
Enrique
E
McQueeney
Kelly D
KD
Jeffers
Mike
M
MacDougall
John
J
Boldog
Ferenc L
FL
Hackett
Craig
C
Shenoy
Suresh
S
Khramtsov
Nikolai
N
Weiner
Jami
J
Lichenstein
Henri S
HS
LaRochelle
William J
WJ
D018262:Adenocarcinoma, Clear Cell / Q000098:blood supply / Q000378:metabolism / Q000628:therapy; D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D002292:Carcinoma, Renal Cell / Q000098:blood supply* / Q000378:metabolism / Q000628:therapy; D002465:Cell Movement / Q000187:drug effects; D003094:Collagen / Q000378:metabolism; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000166:cytology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism; D006801:Humans; D007680:Kidney Neoplasms / Q000098:blood supply / Q000378:metabolism / Q000628:therapy; D007797:Laminin / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D011509:Proteoglycans / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D039961:Semaphorins / Q000235:genetics / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D019061:src-Family Kinases / Q000378:metabolism
D020533:Angiogenesis Inhibitors; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D012333:RNA, Messenger; D011994:Recombinant Proteins; C504201:SEMA6A protein, human; D039961:Semaphorins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2; C059485:matrigel; D003094:Collagen; D051417:Focal Adhesion Kinase 1; C496051:Ptk2 protein, mouse; D019061:src-Family Kinases; D048049:Extracellular Signal-Regulated MAP Kinases