Overcoming the metastasis-enhancing potential of human tumor necrosis factor alpha by introducing the cell-adhesive Arg-Gly-Asp sequence.
A mutein, F4168, of human tumor necrosis factor alpha (hTNF-alpha) containing the cell-adhesive Arg-Gly-Asp (RGD) sequence near the N terminus was constructed. In contrast to hTNF-alpha, the mutein had binding activity to B16F10/L5 melanoma cells similar to that of fibronectin or laminin, indicating that the adhesive nature of the RGD sequence is conferred upon hTNF-alpha. Introduction of the RGD sequence did not alter the antitumor potential of hTNF-alpha. Simultaneous injection of F4168 and B16F10/L5 melanoma cells into mice did not enhance metastasis formation in lungs, whereas hTNF-alpha significantly promoted it. Enhancement of spontaneous lymph node metastasis of B16F10/L5 cells was also evident in TNF-alpha- but not in F4168-treated mice. In the spontaneous lymph node metastasis model of MethA fibrosarcoma, F4168 injection inhibited metastasis formation more effectively than hTNF-alpha. B16F10/L5 melanoma cells treated with hTNF-alpha enhanced not only their binding activity to laminin but also their invasive potential into Matrigel, whereas F4168 showed no such enhancement. These results suggest that F4168 is a low-toxicity mutein of hTNF-alpha.
Miyata
K
K
Medicinal Research Laboratory, Ishihara Sangyo Kaisha, Ltd., Shiga, Japan.
Mitsuishi
Y
Y
Shikama
H
H
Kuroda
K
K
Nishimura
K
K
Sakae
N
N
Kato
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion; D016923:Cell Death; D005354:Fibrosarcoma / Q000556:secondary; D006801:Humans; D008175:Lung Neoplasms / Q000556:secondary; D008207:Lymphatic Metastasis; D008546:Melanoma, Experimental / Q000473:pathology / Q000556:secondary; D051379:Mice; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009362:Neoplasm Metastasis; D009842:Oligopeptides / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000502:physiology*; D012878:Skin Neoplasms / Q000556:secondary; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000502:physiology*
C098256:F 4168; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; C047981:arginyl-glycyl-aspartic acid