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Cycloamylose-nanogel drug delivery system-mediated intratumor silencing of the vascular endothelial growth factor regulates neovascularization in tumor microenvironment.

Fujii H, Shin-Ya M, Takeda S, Hashimoto Y, Mukai SA, Sawada S, Adachi T, Akiyoshi K, Miki T, Mazda O - Cancer Sci. (2014)

Bottom Line: The siVEGF/nanogel complex was engulfed by renal cell carcinoma (RCC) cells through the endocytotic pathway, resulting in efficient knockdown of VEGF.Intra-tumor injections of the complex significantly suppressed neovascularization and growth of RCC in mice.The treatment also inhibited induction of myeloid-derived suppressor cells, while it decreased interleukin-17A production.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Kyoto Prefectural University of Medicine, Kyoto, Kyoto Prefectural University of Medicine, Kyoto, Japan; Department of Urology, Kyoto Prefectural University of Medicine, Kyoto, Japan.

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Vascular endothelial growth factor (VEGF) silencing in tumor significantly suppressed myeloid-derived suppressor cells (MDSC) and interleukin-17A (IL-17A) induction in the spleen. Tumor-bearing mice were given intratumor injections with nanogel alone or siRNA/nanogel complex as in Figure 5. Mice were killed on day 20. (a) Spleen cells were stained with FITC-anti-Gr1 and APC-CD11b antibodies followed by FACS analysis. Numbers in dot plots (upper) indicate proportions of Gr-1+ CD11b+ cells (blue rectangles), while means ± SD (n = at least 7/group) of Gr-1+ CD11b+ populations are plotted (lower). (b) Spleen cells were cultured with interferon (IFN)-γ and LPS, and nitric oxide (NO)-producing cells were visualized by DAF-2 DA. Arrows indicate NO high producer cells. Scale bar, 50 μm. (c) Immunohistochemical staining of the tumor sections with CD11b (upper) and anti-Gr-1 (lower) antibodies. Original magnification was 200×. (d) Spleen cells were cultured with conA, and 24 h later IL-17A concentrations in culture supernatant were measured by the Cytometric Bead Array assay. Data represent the means ± SD (n = 3). *P < 0.05.
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fig06: Vascular endothelial growth factor (VEGF) silencing in tumor significantly suppressed myeloid-derived suppressor cells (MDSC) and interleukin-17A (IL-17A) induction in the spleen. Tumor-bearing mice were given intratumor injections with nanogel alone or siRNA/nanogel complex as in Figure 5. Mice were killed on day 20. (a) Spleen cells were stained with FITC-anti-Gr1 and APC-CD11b antibodies followed by FACS analysis. Numbers in dot plots (upper) indicate proportions of Gr-1+ CD11b+ cells (blue rectangles), while means ± SD (n = at least 7/group) of Gr-1+ CD11b+ populations are plotted (lower). (b) Spleen cells were cultured with interferon (IFN)-γ and LPS, and nitric oxide (NO)-producing cells were visualized by DAF-2 DA. Arrows indicate NO high producer cells. Scale bar, 50 μm. (c) Immunohistochemical staining of the tumor sections with CD11b (upper) and anti-Gr-1 (lower) antibodies. Original magnification was 200×. (d) Spleen cells were cultured with conA, and 24 h later IL-17A concentrations in culture supernatant were measured by the Cytometric Bead Array assay. Data represent the means ± SD (n = 3). *P < 0.05.

Mentions: We assessed the potential influence of intra-tumor VEGF silencing on systemic immunity. To this end, we estimated the accumulation of MDSC in spleens of the nanogel-treated mice. The percentage of splenic CD11b+Gr1+MDSC significantly decreased in the siVEGF/nanogel-treated mice compared with that in siCont/nanogel-treated mice (Fig. 6a). MDSC activity was visualized by diaminofluorescein-2 (DAF-2) assay, and the results revealed that MDSC with high nitric oxide (NO) producing activity(12) were considerably abundant in the control group but not in the siVEGF/nanogel-treated group (Fig. 6b). In addition, we stained the tumor sections with CD11b and anti-Gr-1 antibodies, and found fewer stained cells present in the siVEGF/nanogel treated-tumors than in the control tumors (Fig. 6c). These results strongly suggest that MDSC in spleens and tumors were reduced after the VEGF-silencing using the nanogel.


Cycloamylose-nanogel drug delivery system-mediated intratumor silencing of the vascular endothelial growth factor regulates neovascularization in tumor microenvironment.

Fujii H, Shin-Ya M, Takeda S, Hashimoto Y, Mukai SA, Sawada S, Adachi T, Akiyoshi K, Miki T, Mazda O - Cancer Sci. (2014)

Vascular endothelial growth factor (VEGF) silencing in tumor significantly suppressed myeloid-derived suppressor cells (MDSC) and interleukin-17A (IL-17A) induction in the spleen. Tumor-bearing mice were given intratumor injections with nanogel alone or siRNA/nanogel complex as in Figure 5. Mice were killed on day 20. (a) Spleen cells were stained with FITC-anti-Gr1 and APC-CD11b antibodies followed by FACS analysis. Numbers in dot plots (upper) indicate proportions of Gr-1+ CD11b+ cells (blue rectangles), while means ± SD (n = at least 7/group) of Gr-1+ CD11b+ populations are plotted (lower). (b) Spleen cells were cultured with interferon (IFN)-γ and LPS, and nitric oxide (NO)-producing cells were visualized by DAF-2 DA. Arrows indicate NO high producer cells. Scale bar, 50 μm. (c) Immunohistochemical staining of the tumor sections with CD11b (upper) and anti-Gr-1 (lower) antibodies. Original magnification was 200×. (d) Spleen cells were cultured with conA, and 24 h later IL-17A concentrations in culture supernatant were measured by the Cytometric Bead Array assay. Data represent the means ± SD (n = 3). *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4317968&req=5

fig06: Vascular endothelial growth factor (VEGF) silencing in tumor significantly suppressed myeloid-derived suppressor cells (MDSC) and interleukin-17A (IL-17A) induction in the spleen. Tumor-bearing mice were given intratumor injections with nanogel alone or siRNA/nanogel complex as in Figure 5. Mice were killed on day 20. (a) Spleen cells were stained with FITC-anti-Gr1 and APC-CD11b antibodies followed by FACS analysis. Numbers in dot plots (upper) indicate proportions of Gr-1+ CD11b+ cells (blue rectangles), while means ± SD (n = at least 7/group) of Gr-1+ CD11b+ populations are plotted (lower). (b) Spleen cells were cultured with interferon (IFN)-γ and LPS, and nitric oxide (NO)-producing cells were visualized by DAF-2 DA. Arrows indicate NO high producer cells. Scale bar, 50 μm. (c) Immunohistochemical staining of the tumor sections with CD11b (upper) and anti-Gr-1 (lower) antibodies. Original magnification was 200×. (d) Spleen cells were cultured with conA, and 24 h later IL-17A concentrations in culture supernatant were measured by the Cytometric Bead Array assay. Data represent the means ± SD (n = 3). *P < 0.05.
Mentions: We assessed the potential influence of intra-tumor VEGF silencing on systemic immunity. To this end, we estimated the accumulation of MDSC in spleens of the nanogel-treated mice. The percentage of splenic CD11b+Gr1+MDSC significantly decreased in the siVEGF/nanogel-treated mice compared with that in siCont/nanogel-treated mice (Fig. 6a). MDSC activity was visualized by diaminofluorescein-2 (DAF-2) assay, and the results revealed that MDSC with high nitric oxide (NO) producing activity(12) were considerably abundant in the control group but not in the siVEGF/nanogel-treated group (Fig. 6b). In addition, we stained the tumor sections with CD11b and anti-Gr-1 antibodies, and found fewer stained cells present in the siVEGF/nanogel treated-tumors than in the control tumors (Fig. 6c). These results strongly suggest that MDSC in spleens and tumors were reduced after the VEGF-silencing using the nanogel.

Bottom Line: The siVEGF/nanogel complex was engulfed by renal cell carcinoma (RCC) cells through the endocytotic pathway, resulting in efficient knockdown of VEGF.Intra-tumor injections of the complex significantly suppressed neovascularization and growth of RCC in mice.The treatment also inhibited induction of myeloid-derived suppressor cells, while it decreased interleukin-17A production.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Kyoto Prefectural University of Medicine, Kyoto, Kyoto Prefectural University of Medicine, Kyoto, Japan; Department of Urology, Kyoto Prefectural University of Medicine, Kyoto, Japan.

Show MeSH
Related in: MedlinePlus