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MT95-4, a fully humanized antibody raised against aminopeptidase N, reduces tumor progression in a mouse model.

Akita S, Hattori N, Masuda T, Horimasu Y, Nakashima T, Iwamoto H, Fujitaka K, Miyake M, Kohno N - Cancer Sci. (2015)

Bottom Line: To develop a novel monoclonal antibody-based cancer therapy targeting APN/CD13, we established a fully humanized anti-APN/CD13 monoclonal antibody, MT95-4.We found that expression of human APN/CD13 in murine melanoma cells increased the size of subcutaneous tumors, extent of lung metastasis and degree of angiogenesis in the subcutaneous tumors; these tumor-promoting and angiogenesis-promoting characteristics were reduced by the i.p. administration of MT95-4.These results suggested that the antitumor and anti-angiogenic effects of MT95-4 were dependent on APN/CD13 expression in tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Internal Medicine, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

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Effects of MT95-4 on tumor progression in the tail vein metastasis model and subcutaneous tumor model using human lung cancer cells. Mice were administered MT95-4 or control IgG (1 mg/kg) i.p. twice a week. (a) Expression levels of APN/CD13 mRNA in human lung cancer cells analyzed by real-time PCR. (b) Expression levels of APN/CD13 on the surface of human lung cancer cells, as analyzed by flow cytometry. (c), (f) and (i) Evaluation of tumor volumes in a subcutaneous tumor model using H1299 (c), PC14 (f) and A549 (i) cells. The sizes of subcutaneous tumors were measured once a week for 5 or 6 weeks after inoculation of tumor cells. The data represent the means (± SEM) for eight mice per group. $P < 0.05; NS, not significant. (d), (g) and (j) Evaluation of angiogenesis in subcutaneous tumors derived from H1299 (d), PC14 (g) and A549 (j) cells. The area of CD31-positive vessels was measured. Data represent the means (± SEM) for eight mice per group. $$$P < 0.001; NS, not significant. (e), (h) and (k) Immunohistochemical staining for CD31 in a subcutaneous tumor. Scale bar, 100 μm.
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fig04: Effects of MT95-4 on tumor progression in the tail vein metastasis model and subcutaneous tumor model using human lung cancer cells. Mice were administered MT95-4 or control IgG (1 mg/kg) i.p. twice a week. (a) Expression levels of APN/CD13 mRNA in human lung cancer cells analyzed by real-time PCR. (b) Expression levels of APN/CD13 on the surface of human lung cancer cells, as analyzed by flow cytometry. (c), (f) and (i) Evaluation of tumor volumes in a subcutaneous tumor model using H1299 (c), PC14 (f) and A549 (i) cells. The sizes of subcutaneous tumors were measured once a week for 5 or 6 weeks after inoculation of tumor cells. The data represent the means (± SEM) for eight mice per group. $P < 0.05; NS, not significant. (d), (g) and (j) Evaluation of angiogenesis in subcutaneous tumors derived from H1299 (d), PC14 (g) and A549 (j) cells. The area of CD31-positive vessels was measured. Data represent the means (± SEM) for eight mice per group. $$$P < 0.001; NS, not significant. (e), (h) and (k) Immunohistochemical staining for CD31 in a subcutaneous tumor. Scale bar, 100 μm.

Mentions: To further verify whether the antitumor effects of MT95-4 were associated with the expression of APN/CD13 in cancer cells, we administered MT95-4 to mice bearing human lung cancer cells abundantly or weakly expressing APN/CD13. As shown in Figure4a and b, real-time PCR analysis and flow cytometric analysis revealed that APN/CD13 was abundantly expressed in H1299 and PC14 cells but weakly expressed in A549 cells. Thus, we established subcutaneous tumor xenografts using H1299, PC14 and A549 cells in NOD/SCID mice, and MT95-4 or control IgG was administered i.p. The administration of MT95-4 reduced tumor volumes in mice bearing H1299 and PC14 cells (Fig.4c,f) but not in mice bearing A549 cells (Fig.4i). Moreover, the degree of angiogenesis in subcutaneous tumors derived from H1299 and PC14 cells was lower in the MT95-4-treated group than in the control group (Fig.4d,g); however, there were no differences in the degree of angiogenesis between MT95-4-treated and control IgG-treated mice bearing tumors derived from A549 cells (Fig.4j).


MT95-4, a fully humanized antibody raised against aminopeptidase N, reduces tumor progression in a mouse model.

Akita S, Hattori N, Masuda T, Horimasu Y, Nakashima T, Iwamoto H, Fujitaka K, Miyake M, Kohno N - Cancer Sci. (2015)

Effects of MT95-4 on tumor progression in the tail vein metastasis model and subcutaneous tumor model using human lung cancer cells. Mice were administered MT95-4 or control IgG (1 mg/kg) i.p. twice a week. (a) Expression levels of APN/CD13 mRNA in human lung cancer cells analyzed by real-time PCR. (b) Expression levels of APN/CD13 on the surface of human lung cancer cells, as analyzed by flow cytometry. (c), (f) and (i) Evaluation of tumor volumes in a subcutaneous tumor model using H1299 (c), PC14 (f) and A549 (i) cells. The sizes of subcutaneous tumors were measured once a week for 5 or 6 weeks after inoculation of tumor cells. The data represent the means (± SEM) for eight mice per group. $P < 0.05; NS, not significant. (d), (g) and (j) Evaluation of angiogenesis in subcutaneous tumors derived from H1299 (d), PC14 (g) and A549 (j) cells. The area of CD31-positive vessels was measured. Data represent the means (± SEM) for eight mice per group. $$$P < 0.001; NS, not significant. (e), (h) and (k) Immunohistochemical staining for CD31 in a subcutaneous tumor. Scale bar, 100 μm.
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fig04: Effects of MT95-4 on tumor progression in the tail vein metastasis model and subcutaneous tumor model using human lung cancer cells. Mice were administered MT95-4 or control IgG (1 mg/kg) i.p. twice a week. (a) Expression levels of APN/CD13 mRNA in human lung cancer cells analyzed by real-time PCR. (b) Expression levels of APN/CD13 on the surface of human lung cancer cells, as analyzed by flow cytometry. (c), (f) and (i) Evaluation of tumor volumes in a subcutaneous tumor model using H1299 (c), PC14 (f) and A549 (i) cells. The sizes of subcutaneous tumors were measured once a week for 5 or 6 weeks after inoculation of tumor cells. The data represent the means (± SEM) for eight mice per group. $P < 0.05; NS, not significant. (d), (g) and (j) Evaluation of angiogenesis in subcutaneous tumors derived from H1299 (d), PC14 (g) and A549 (j) cells. The area of CD31-positive vessels was measured. Data represent the means (± SEM) for eight mice per group. $$$P < 0.001; NS, not significant. (e), (h) and (k) Immunohistochemical staining for CD31 in a subcutaneous tumor. Scale bar, 100 μm.
Mentions: To further verify whether the antitumor effects of MT95-4 were associated with the expression of APN/CD13 in cancer cells, we administered MT95-4 to mice bearing human lung cancer cells abundantly or weakly expressing APN/CD13. As shown in Figure4a and b, real-time PCR analysis and flow cytometric analysis revealed that APN/CD13 was abundantly expressed in H1299 and PC14 cells but weakly expressed in A549 cells. Thus, we established subcutaneous tumor xenografts using H1299, PC14 and A549 cells in NOD/SCID mice, and MT95-4 or control IgG was administered i.p. The administration of MT95-4 reduced tumor volumes in mice bearing H1299 and PC14 cells (Fig.4c,f) but not in mice bearing A549 cells (Fig.4i). Moreover, the degree of angiogenesis in subcutaneous tumors derived from H1299 and PC14 cells was lower in the MT95-4-treated group than in the control group (Fig.4d,g); however, there were no differences in the degree of angiogenesis between MT95-4-treated and control IgG-treated mice bearing tumors derived from A549 cells (Fig.4j).

Bottom Line: To develop a novel monoclonal antibody-based cancer therapy targeting APN/CD13, we established a fully humanized anti-APN/CD13 monoclonal antibody, MT95-4.We found that expression of human APN/CD13 in murine melanoma cells increased the size of subcutaneous tumors, extent of lung metastasis and degree of angiogenesis in the subcutaneous tumors; these tumor-promoting and angiogenesis-promoting characteristics were reduced by the i.p. administration of MT95-4.These results suggested that the antitumor and anti-angiogenic effects of MT95-4 were dependent on APN/CD13 expression in tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Internal Medicine, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

Show MeSH
Related in: MedlinePlus