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Oncolytic Activity of a Recombinant Measles Virus, Blind to Signaling Lymphocyte Activation Molecule, Against Colorectal Cancer Cells.

Amagai Y, Fujiyuki T, Yoneda M, Shoji K, Furukawa Y, Sato H, Kai C - Sci Rep (2016)

Bottom Line: Oncolytic virotherapy is a distinctive antitumor therapy based on the cancer-cell-specific infectivity and killing activity of viruses, which exert a considerable antitumor effect with only a few treatments.Tumour progression in xenograft models was also abrogated by the virus, and the infection of cancer cells in vivo by the virus was demonstrated with both flow cytometry and a histological analysis.Therefore, rMV-SLAMblind is considered a novel therapeutic agent for colorectal cancers, including those resistant to molecular-targeted therapies.

View Article: PubMed Central - PubMed

Affiliation: Laboratory Animal Research Center, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.

ABSTRACT
Oncolytic virotherapy is a distinctive antitumor therapy based on the cancer-cell-specific infectivity and killing activity of viruses, which exert a considerable antitumor effect with only a few treatments. Because colorectal cancer cells often acquire resistance to the molecular-targeted therapies and alternative treatments are called for, in this study, we evaluated the oncolytic activity against colorectal cancer cells of a recombinant measles virus (rMV-SLAMblind), which is blind to signaling lymphocytic activation molecule (SLAM) and infects target cells via nectin-4/poliovirus receptor-related 4 protein. We examined 10 cell lines including 8 cell lines that were resistant to epidermal-growth-factor-receptor (EGFR) targeted therapy. rMV-SLAMblind infected and lysed the nectin-4-positive cell lines dependently on nectin-4 expression, in spite of mutation in EGFR cascade. Tumour progression in xenograft models was also abrogated by the virus, and the infection of cancer cells in vivo by the virus was demonstrated with both flow cytometry and a histological analysis. Therefore, rMV-SLAMblind is considered a novel therapeutic agent for colorectal cancers, including those resistant to molecular-targeted therapies.

No MeSH data available.


Related in: MedlinePlus

Antitumor effects of rMV-EGFP-SLAMblind in vivo.Growth curves of DLD1 (a) and HT29 cells (b) in vivo. A total number of 106 DLD1 and HT29 cells were injected subcutaneously into seven SCID mice each, and the tumour sizes were measured every 2 or 3 days. Arrows indicate the days upon which the mice were injected intratumorally with rMV-EGFP-SLAMblind at 106 TCID50/mouse (days 0, 7, and 14). Each datum represents a mean ± SD (n = 7). *p < 0.05, **p < 0.01 compared with the vehicle-treated control on Welch’s t test. (c) Antitumor effects conferred by rMV-EGFP-SLAMblind. All mice were euthanized on day 20 and their tumours weighed. The bar indicates the mean average for each group. (n = 7). **p < 0.01 compared with the vehicle-treated controls on Welch’s t test. (d) Representative photographs of a tumour in each treatment group. (e) Flow-cytometric analysis of live virus-infected tumour cells. All plots are density plots and fluorescent intensities were plotted on bi-exponential axes. (f) The plots indicate the proportion of EGFP-positive cells in each mouse (n = 7 in each group), and the bar indicates the mean for each group. **p < 0.01 compared with the vehicle-treated control on Welch’s t test.
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f3: Antitumor effects of rMV-EGFP-SLAMblind in vivo.Growth curves of DLD1 (a) and HT29 cells (b) in vivo. A total number of 106 DLD1 and HT29 cells were injected subcutaneously into seven SCID mice each, and the tumour sizes were measured every 2 or 3 days. Arrows indicate the days upon which the mice were injected intratumorally with rMV-EGFP-SLAMblind at 106 TCID50/mouse (days 0, 7, and 14). Each datum represents a mean ± SD (n = 7). *p < 0.05, **p < 0.01 compared with the vehicle-treated control on Welch’s t test. (c) Antitumor effects conferred by rMV-EGFP-SLAMblind. All mice were euthanized on day 20 and their tumours weighed. The bar indicates the mean average for each group. (n = 7). **p < 0.01 compared with the vehicle-treated controls on Welch’s t test. (d) Representative photographs of a tumour in each treatment group. (e) Flow-cytometric analysis of live virus-infected tumour cells. All plots are density plots and fluorescent intensities were plotted on bi-exponential axes. (f) The plots indicate the proportion of EGFP-positive cells in each mouse (n = 7 in each group), and the bar indicates the mean for each group. **p < 0.01 compared with the vehicle-treated control on Welch’s t test.

Mentions: The antitumor effects of rMV-EGFP-SLAMblind in vivo were examined using xenograft models. DLD1 and HT29 cells were transplanted into C.B-17/Icr-scid/scidJcl (SCID) mice. When the tumours reached 200 mm3, they were inoculated with rMV-EGFP-SLAMblind three times at weekly intervals. As shown in Fig. 3a, the administration of rMV-EGFP-SLAMblind exerted potent antitumor effects on the DLD1 cells, resulting in a reduction in the tumour volume of approximately 55% compared with the control. Similar results were obtained with the HT29 cell transplantation model, resulting in a reduction in the tumour volume of approximately 60% compared with the control (Fig. 3b). Twenty days after the first inoculation, each mouse was euthanized and their tumours weighed. The mean tumour weight in the virus-treated group was significantly lower than that in the control group for each type of tumour (Fig. 3c,d). We also performed a flow-cytometric analysis to investigate whether rMV-EGFP-SLAMblind remained within the tumour a week after the last administration of the virus. The live cell population was selected based on the incorporation of 7-amino-actinomycin D (7-AAD), detected with forward/side scatter (FSC/SSC; Fig. 3e). The mouse-derived H2Kd-positive cells were gated out to focus on the live tumour cells (Fig. 3e). The proportion of EGFP-positive cells within the gate, which represented the rMV-EGFP-SLAMblind-infected live tumour cells, was 1–6% (2.9% on average) in the DLD1 cells and 0.2–2% (1.0% on average) in the HT29 cells (Fig. 3e,f). In contrast, the tumours in the control group were negative for EGFP (Fig. 3f). To analyse the distribution of the virus-infected cells, a histopathological analysis was performed on tumour tissues treated with rMV-EGFP-SLAMblind. Large necrotic regions were observed histopathologically in the tumour masses from both xenograft models, established with DLD1 and HT29 cells (Fig. 4a, H–E). Positive EGFP fluorescence, indicating the presence of the virus in growing cells, was observed in the area adjacent to the necrotic region (Fig. 4b). MV-N protein was immunostained in an analysis of serial sections, and the MV-N-positive area corresponded to the necrotic region and the EGFP-positive area in both xenograft models (DLD1 and HT29 cells) (Fig. 4a,b).


Oncolytic Activity of a Recombinant Measles Virus, Blind to Signaling Lymphocyte Activation Molecule, Against Colorectal Cancer Cells.

Amagai Y, Fujiyuki T, Yoneda M, Shoji K, Furukawa Y, Sato H, Kai C - Sci Rep (2016)

Antitumor effects of rMV-EGFP-SLAMblind in vivo.Growth curves of DLD1 (a) and HT29 cells (b) in vivo. A total number of 106 DLD1 and HT29 cells were injected subcutaneously into seven SCID mice each, and the tumour sizes were measured every 2 or 3 days. Arrows indicate the days upon which the mice were injected intratumorally with rMV-EGFP-SLAMblind at 106 TCID50/mouse (days 0, 7, and 14). Each datum represents a mean ± SD (n = 7). *p < 0.05, **p < 0.01 compared with the vehicle-treated control on Welch’s t test. (c) Antitumor effects conferred by rMV-EGFP-SLAMblind. All mice were euthanized on day 20 and their tumours weighed. The bar indicates the mean average for each group. (n = 7). **p < 0.01 compared with the vehicle-treated controls on Welch’s t test. (d) Representative photographs of a tumour in each treatment group. (e) Flow-cytometric analysis of live virus-infected tumour cells. All plots are density plots and fluorescent intensities were plotted on bi-exponential axes. (f) The plots indicate the proportion of EGFP-positive cells in each mouse (n = 7 in each group), and the bar indicates the mean for each group. **p < 0.01 compared with the vehicle-treated control on Welch’s t test.
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Related In: Results  -  Collection

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f3: Antitumor effects of rMV-EGFP-SLAMblind in vivo.Growth curves of DLD1 (a) and HT29 cells (b) in vivo. A total number of 106 DLD1 and HT29 cells were injected subcutaneously into seven SCID mice each, and the tumour sizes were measured every 2 or 3 days. Arrows indicate the days upon which the mice were injected intratumorally with rMV-EGFP-SLAMblind at 106 TCID50/mouse (days 0, 7, and 14). Each datum represents a mean ± SD (n = 7). *p < 0.05, **p < 0.01 compared with the vehicle-treated control on Welch’s t test. (c) Antitumor effects conferred by rMV-EGFP-SLAMblind. All mice were euthanized on day 20 and their tumours weighed. The bar indicates the mean average for each group. (n = 7). **p < 0.01 compared with the vehicle-treated controls on Welch’s t test. (d) Representative photographs of a tumour in each treatment group. (e) Flow-cytometric analysis of live virus-infected tumour cells. All plots are density plots and fluorescent intensities were plotted on bi-exponential axes. (f) The plots indicate the proportion of EGFP-positive cells in each mouse (n = 7 in each group), and the bar indicates the mean for each group. **p < 0.01 compared with the vehicle-treated control on Welch’s t test.
Mentions: The antitumor effects of rMV-EGFP-SLAMblind in vivo were examined using xenograft models. DLD1 and HT29 cells were transplanted into C.B-17/Icr-scid/scidJcl (SCID) mice. When the tumours reached 200 mm3, they were inoculated with rMV-EGFP-SLAMblind three times at weekly intervals. As shown in Fig. 3a, the administration of rMV-EGFP-SLAMblind exerted potent antitumor effects on the DLD1 cells, resulting in a reduction in the tumour volume of approximately 55% compared with the control. Similar results were obtained with the HT29 cell transplantation model, resulting in a reduction in the tumour volume of approximately 60% compared with the control (Fig. 3b). Twenty days after the first inoculation, each mouse was euthanized and their tumours weighed. The mean tumour weight in the virus-treated group was significantly lower than that in the control group for each type of tumour (Fig. 3c,d). We also performed a flow-cytometric analysis to investigate whether rMV-EGFP-SLAMblind remained within the tumour a week after the last administration of the virus. The live cell population was selected based on the incorporation of 7-amino-actinomycin D (7-AAD), detected with forward/side scatter (FSC/SSC; Fig. 3e). The mouse-derived H2Kd-positive cells were gated out to focus on the live tumour cells (Fig. 3e). The proportion of EGFP-positive cells within the gate, which represented the rMV-EGFP-SLAMblind-infected live tumour cells, was 1–6% (2.9% on average) in the DLD1 cells and 0.2–2% (1.0% on average) in the HT29 cells (Fig. 3e,f). In contrast, the tumours in the control group were negative for EGFP (Fig. 3f). To analyse the distribution of the virus-infected cells, a histopathological analysis was performed on tumour tissues treated with rMV-EGFP-SLAMblind. Large necrotic regions were observed histopathologically in the tumour masses from both xenograft models, established with DLD1 and HT29 cells (Fig. 4a, H–E). Positive EGFP fluorescence, indicating the presence of the virus in growing cells, was observed in the area adjacent to the necrotic region (Fig. 4b). MV-N protein was immunostained in an analysis of serial sections, and the MV-N-positive area corresponded to the necrotic region and the EGFP-positive area in both xenograft models (DLD1 and HT29 cells) (Fig. 4a,b).

Bottom Line: Oncolytic virotherapy is a distinctive antitumor therapy based on the cancer-cell-specific infectivity and killing activity of viruses, which exert a considerable antitumor effect with only a few treatments.Tumour progression in xenograft models was also abrogated by the virus, and the infection of cancer cells in vivo by the virus was demonstrated with both flow cytometry and a histological analysis.Therefore, rMV-SLAMblind is considered a novel therapeutic agent for colorectal cancers, including those resistant to molecular-targeted therapies.

View Article: PubMed Central - PubMed

Affiliation: Laboratory Animal Research Center, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.

ABSTRACT
Oncolytic virotherapy is a distinctive antitumor therapy based on the cancer-cell-specific infectivity and killing activity of viruses, which exert a considerable antitumor effect with only a few treatments. Because colorectal cancer cells often acquire resistance to the molecular-targeted therapies and alternative treatments are called for, in this study, we evaluated the oncolytic activity against colorectal cancer cells of a recombinant measles virus (rMV-SLAMblind), which is blind to signaling lymphocytic activation molecule (SLAM) and infects target cells via nectin-4/poliovirus receptor-related 4 protein. We examined 10 cell lines including 8 cell lines that were resistant to epidermal-growth-factor-receptor (EGFR) targeted therapy. rMV-SLAMblind infected and lysed the nectin-4-positive cell lines dependently on nectin-4 expression, in spite of mutation in EGFR cascade. Tumour progression in xenograft models was also abrogated by the virus, and the infection of cancer cells in vivo by the virus was demonstrated with both flow cytometry and a histological analysis. Therefore, rMV-SLAMblind is considered a novel therapeutic agent for colorectal cancers, including those resistant to molecular-targeted therapies.

No MeSH data available.


Related in: MedlinePlus