GD1alpha-replica peptides functionally mimic GD1alpha, an adhesion molecule of metastatic tumor cells, and suppress the tumor metastasis.
A novel peptide technology to produce mimicking peptides of carbohydrate moiety (which we propose to name glyco-replica peptides) is a useful tool to elucidate the functions of glycoconjugate. Carbohydrate moiety of ganglioside GD1alpha functions as a molecule involved in the adhesion between murine highly metastatic lymphoma RAW117-H10 cells and hepatic sinusoidal endothelial (HSE) cells. To prepare peptides which mimic the carbohydrate structure of GD1alpha, phage clones expressing peptides which bound to a monoclonal antibody against GD1alpha (KA17) were isolated from a phage-displayed random peptide library. Four phage clones having affinity to the monoclonal antibody KA17 were isolated, and these clones showed inhibitory effect on the binding of KA17 to GD1alpha. The amino acid sequences of the displayed pentadecamers were determined, and one of the phages displaying sequence WHWRHRIPLQLAAGR bound to HSE cells directly and showed the highest inhibitory effect on the adhesion between RAW117-H10 cells and HSE cells. The synthesized peptides having the same sequences to the displayed 15mers in the four isolated phage clones also showed the inhibitory effect on the adhesion of RAW117-H10 cells to HSE cells, and, again, the WHWRHRIPLQLAAGR peptide showed the highest inhibitory effect. Furthermore, intravenous injection of the peptide brought almost complete inhibition of the metastasis of RAW117-H10 cells to lung and spleen, and about 50% inhibition of the liver metastasis. These results indicate that GD1alpha plays an important role for metastasis of RAW117-H10 cells, and the peptides obtained by the present procedure are able to mimic the functional role of the glycoconjugate.
Ishikawa
D
D
Cellular Technology Institute, Otsuka Pharmaceutical Co. Ltd., Tokushima, Japan.
Kikkawa
H
H
Ogino
K
K
Hirabayashi
Y
Y
Oku
N
N
Taki
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001435:Bacteriophages; D002448:Cell Adhesion / Q000187:drug effects*; D002460:Cell Line; D000939:Epitopes / Q000737:chemistry; D005260:Female; D005677:G(M1) Ganglioside / Q000031:analogs & derivatives* / Q000502:physiology / Q000633:toxicity; D008113:Liver Neoplasms / Q000473:pathology / Q000517:prevention & control / Q000556:secondary; D008175:Lung Neoplasms / Q000473:pathology / Q000517:prevention & control / Q000556:secondary; D008223:Lymphoma / Q000188:drug therapy / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis / Q000517:prevention & control*; D019151:Peptide Library; D010455:Peptides / Q000633:toxicity*; D013160:Splenic Neoplasms / Q000473:pathology / Q000517:prevention & control / Q000556:secondary; D014407:Tumor Cells, Cultured
D000939:Epitopes; D019151:Peptide Library; D010455:Peptides; C064020:ganglioside GD1alpha; D005677:G(M1) Ganglioside