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Anti-Neuroblastoma Activity of Gold Nanorods Bound with GD2 Monoclonal Antibody under Near-Infrared Laser Irradiation.

Peng CA, Wang CH - Cancers (Basel) (2011)

Bottom Line: Our results illustrated that anti-GD2-conjugated CGNRs were extensively endocytosed by GD2+ stNB-V1 neuroblastoma cells via antibody-mediated endocytosis.In addition, we showed that anti-GD2 bound CGNRs were not internalized by GD2- SH-SY5Y neuroblastoma cells.Based on the in vitro study, CGNRs bound with anti-GD2 has the potential to be utilized as a therapeutic thermal coupling agent that generates heat sufficient to selectively kill neuroblastoma cells under NIR laser light exposure.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical Engineering, Michigan Technological University, 1400 Townsend Drive, Houghton, MI 49931, USA. cpeng@mtu.edu.

ABSTRACT
High-risk neuroblastoma is one of the most common deaths in pediatric oncology. Current treatment of this disease involves a coordinated sequence of chemotherapy, surgery, and radiation. Further advances in therapy will require the targeting of tumor cells in a more selective and efficient way so that survival can be improved without substantially increasing toxicity. To achieve tumor-selective delivery, disialoganglioside (GD2) expressed by almost all neuroblastoma tumors represents a potential molecular target that can be exploited for tumor-selective delivery. In this study, GD2 monoclonal antibody (anti-GD2) was conjugated to gold nanorods (GNRs) which are one of anisotropic nanomaterials that can absorb near-infrared (NIR) laser light and convert it to energy for photothermolysis of tumor cells. Thiolated chitosan, due to its biocompatibility, was used to replace cetyltrimethylammonium bromide (CTAB) originally used in the synthesis of gold nanorods. In order to specifically target GD2 overexpressed on the surface of neuroblastoma stNB-V1 cells, anti-GD2 was conjugated to chitosan modified GNRs (CGNRs). To examine the fate of CGNRs conjugated with anti-GD2 after incubation with neuroblastoma cells, rhadoamine B was labeled on CGNRs functionalized with anti-GD2. Our results illustrated that anti-GD2-conjugated CGNRs were extensively endocytosed by GD2+ stNB-V1 neuroblastoma cells via antibody-mediated endocytosis. In addition, we showed that anti-GD2 bound CGNRs were not internalized by GD2- SH-SY5Y neuroblastoma cells. After anti-GD2-linked CGNRs were incubated with neuroblatoma cells for six hours, the treated cells were further irradiated with 808 nm NIR laser. Post-NIR laser exposure, when examined by calcein-AM dye, stNB-V1 cells all underwent necrosis, while non-GD2 expressing SH-SY5Y cells all remained viable. Based on the in vitro study, CGNRs bound with anti-GD2 has the potential to be utilized as a therapeutic thermal coupling agent that generates heat sufficient to selectively kill neuroblastoma cells under NIR laser light exposure.

No MeSH data available.


Related in: MedlinePlus

Photothermal treatment of SH-SY5Y (a-c) and stNB-V1 (d-f) cells. After SH-SY5Y and stNB-V1 were incubated with anti-GD2-CGNRs for 6 h, 808 nm NIR laser was harnessed to beam the cells from 0.2 to 2 W/cm2 within 10 min and then maintaining at 2 W/cm2 for an additional 5 min. After staining with 2.5 μM calcein-AM dye, fluorescent images of cells were taken (a, d) within, (b, e) on the edge of, and (c, f) outside the NIR laser-shining zone (scale bar = 100 μm). Green fluorescence indicates viable cells, in contrast to dead cells which reveal no fluorescence.
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f8-cancers-03-00227: Photothermal treatment of SH-SY5Y (a-c) and stNB-V1 (d-f) cells. After SH-SY5Y and stNB-V1 were incubated with anti-GD2-CGNRs for 6 h, 808 nm NIR laser was harnessed to beam the cells from 0.2 to 2 W/cm2 within 10 min and then maintaining at 2 W/cm2 for an additional 5 min. After staining with 2.5 μM calcein-AM dye, fluorescent images of cells were taken (a, d) within, (b, e) on the edge of, and (c, f) outside the NIR laser-shining zone (scale bar = 100 μm). Green fluorescence indicates viable cells, in contrast to dead cells which reveal no fluorescence.

Mentions: Photothermal treatment was first carried out separately for GD2+ stNB-V1 and GD2− SH-SY5Y cells 6h post-incubation with 0.1 mM anti-GD2-CGNRs. After staining with calcein-AM dye, 496 nm blue light was used to illuminate cells. In this way, if a cell is viable, it emits green fluorescence, whereas a dead cell will not emit any fluorescent light. After incubation with anti-GD2-CGNRs for 6 h, the fluorescent images of SH-SY5Y cells located in the NIR laser-beaming zone, on the laser-shining edge and far from the beam zone were taken and are shown in Figure 8(a)–(c), respectively. Apparently, the GD2− cells located in all of the three zones remained viable, as indicated by their green fluorescence. However, for GD2+ cells treated with anti-GD2-CGNRs for 6 h and then irradiated with NIR laser light, the cells located in the laser-shining zone (Figure 8(d)) did not reveal green fluorescence (i.e., cell necrosis), compared with the cells harbored far away from the beam zone (Figure 8(f)) yielding green fluorescence indicating cell viability. Figure 8(e) represents the cells located on the edge of the NIR laser irradiation, clearly illustrating the boundary between necrotic (dark) and viable cells (green).


Anti-Neuroblastoma Activity of Gold Nanorods Bound with GD2 Monoclonal Antibody under Near-Infrared Laser Irradiation.

Peng CA, Wang CH - Cancers (Basel) (2011)

Photothermal treatment of SH-SY5Y (a-c) and stNB-V1 (d-f) cells. After SH-SY5Y and stNB-V1 were incubated with anti-GD2-CGNRs for 6 h, 808 nm NIR laser was harnessed to beam the cells from 0.2 to 2 W/cm2 within 10 min and then maintaining at 2 W/cm2 for an additional 5 min. After staining with 2.5 μM calcein-AM dye, fluorescent images of cells were taken (a, d) within, (b, e) on the edge of, and (c, f) outside the NIR laser-shining zone (scale bar = 100 μm). Green fluorescence indicates viable cells, in contrast to dead cells which reveal no fluorescence.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3756358&req=5

f8-cancers-03-00227: Photothermal treatment of SH-SY5Y (a-c) and stNB-V1 (d-f) cells. After SH-SY5Y and stNB-V1 were incubated with anti-GD2-CGNRs for 6 h, 808 nm NIR laser was harnessed to beam the cells from 0.2 to 2 W/cm2 within 10 min and then maintaining at 2 W/cm2 for an additional 5 min. After staining with 2.5 μM calcein-AM dye, fluorescent images of cells were taken (a, d) within, (b, e) on the edge of, and (c, f) outside the NIR laser-shining zone (scale bar = 100 μm). Green fluorescence indicates viable cells, in contrast to dead cells which reveal no fluorescence.
Mentions: Photothermal treatment was first carried out separately for GD2+ stNB-V1 and GD2− SH-SY5Y cells 6h post-incubation with 0.1 mM anti-GD2-CGNRs. After staining with calcein-AM dye, 496 nm blue light was used to illuminate cells. In this way, if a cell is viable, it emits green fluorescence, whereas a dead cell will not emit any fluorescent light. After incubation with anti-GD2-CGNRs for 6 h, the fluorescent images of SH-SY5Y cells located in the NIR laser-beaming zone, on the laser-shining edge and far from the beam zone were taken and are shown in Figure 8(a)–(c), respectively. Apparently, the GD2− cells located in all of the three zones remained viable, as indicated by their green fluorescence. However, for GD2+ cells treated with anti-GD2-CGNRs for 6 h and then irradiated with NIR laser light, the cells located in the laser-shining zone (Figure 8(d)) did not reveal green fluorescence (i.e., cell necrosis), compared with the cells harbored far away from the beam zone (Figure 8(f)) yielding green fluorescence indicating cell viability. Figure 8(e) represents the cells located on the edge of the NIR laser irradiation, clearly illustrating the boundary between necrotic (dark) and viable cells (green).

Bottom Line: Our results illustrated that anti-GD2-conjugated CGNRs were extensively endocytosed by GD2+ stNB-V1 neuroblastoma cells via antibody-mediated endocytosis.In addition, we showed that anti-GD2 bound CGNRs were not internalized by GD2- SH-SY5Y neuroblastoma cells.Based on the in vitro study, CGNRs bound with anti-GD2 has the potential to be utilized as a therapeutic thermal coupling agent that generates heat sufficient to selectively kill neuroblastoma cells under NIR laser light exposure.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical Engineering, Michigan Technological University, 1400 Townsend Drive, Houghton, MI 49931, USA. cpeng@mtu.edu.

ABSTRACT
High-risk neuroblastoma is one of the most common deaths in pediatric oncology. Current treatment of this disease involves a coordinated sequence of chemotherapy, surgery, and radiation. Further advances in therapy will require the targeting of tumor cells in a more selective and efficient way so that survival can be improved without substantially increasing toxicity. To achieve tumor-selective delivery, disialoganglioside (GD2) expressed by almost all neuroblastoma tumors represents a potential molecular target that can be exploited for tumor-selective delivery. In this study, GD2 monoclonal antibody (anti-GD2) was conjugated to gold nanorods (GNRs) which are one of anisotropic nanomaterials that can absorb near-infrared (NIR) laser light and convert it to energy for photothermolysis of tumor cells. Thiolated chitosan, due to its biocompatibility, was used to replace cetyltrimethylammonium bromide (CTAB) originally used in the synthesis of gold nanorods. In order to specifically target GD2 overexpressed on the surface of neuroblastoma stNB-V1 cells, anti-GD2 was conjugated to chitosan modified GNRs (CGNRs). To examine the fate of CGNRs conjugated with anti-GD2 after incubation with neuroblastoma cells, rhadoamine B was labeled on CGNRs functionalized with anti-GD2. Our results illustrated that anti-GD2-conjugated CGNRs were extensively endocytosed by GD2+ stNB-V1 neuroblastoma cells via antibody-mediated endocytosis. In addition, we showed that anti-GD2 bound CGNRs were not internalized by GD2- SH-SY5Y neuroblastoma cells. After anti-GD2-linked CGNRs were incubated with neuroblatoma cells for six hours, the treated cells were further irradiated with 808 nm NIR laser. Post-NIR laser exposure, when examined by calcein-AM dye, stNB-V1 cells all underwent necrosis, while non-GD2 expressing SH-SY5Y cells all remained viable. Based on the in vitro study, CGNRs bound with anti-GD2 has the potential to be utilized as a therapeutic thermal coupling agent that generates heat sufficient to selectively kill neuroblastoma cells under NIR laser light exposure.

No MeSH data available.


Related in: MedlinePlus