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Bioluminescence-activated deep-tissue photodynamic therapy of cancer.

Kim YR, Kim S, Choi JW, Choi SY, Lee SH, Kim H, Hahn SK, Koh GY, Yun SH - Theranostics (2015)

Bottom Line: Owing to the shallow penetration of light in tissues, however, the clinical applications of light-activated therapies have been limited.For monolayer cell culture in vitro incubated with Chlorin e6, BRET energy of about 1 nJ per cell generated as strong cytotoxicity as red laser light irradiation at 2.2 mW/cm(2) for 180 s.Our results show the promising potential of novel bioluminescence-activated PDT.

View Article: PubMed Central - PubMed

Affiliation: 1. Graduate School of Nanoscience and Technology (WCU), Korea Advanced Institute of Science and Technology, 291 Daehak-Ro, Yusong-Gu, Daejon 305-701, Korea ; 2. Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, 291 Daehak-Ro, Yusong-Gu, Daejon 305-701, Korea ; 3. Department of Oncology, Asan Medical Center, Univ. Ulsan College of Medicine, Seoul , Korea.

ABSTRACT
Optical energy can trigger a variety of photochemical processes useful for therapies. Owing to the shallow penetration of light in tissues, however, the clinical applications of light-activated therapies have been limited. Bioluminescence resonant energy transfer (BRET) may provide a new way of inducing photochemical activation. Here, we show that efficient bioluminescence energy-induced photodynamic therapy (PDT) of macroscopic tumors and metastases in deep tissue. For monolayer cell culture in vitro incubated with Chlorin e6, BRET energy of about 1 nJ per cell generated as strong cytotoxicity as red laser light irradiation at 2.2 mW/cm(2) for 180 s. Regional delivery of bioluminescence agents via draining lymphatic vessels killed tumor cells spread to the sentinel and secondary lymph nodes, reduced distant metastases in the lung and improved animal survival. Our results show the promising potential of novel bioluminescence-activated PDT.

No MeSH data available.


Related in: MedlinePlus

Treatment of lymph-node metastases. a, BL image of a C57/BL6 mouse with B16F10 tumor in the footpad (day 21), after injection of Luc-QD and CTZ into the foot. b, IHC images of p-LN sections harvested from an untreated mouse at day 23, stained for blood vessels (CD31), lymphatics (VEGFR3/CD31), melanoma cells (melan-A), and apoptosis (caspase-3). Strong green autofluorescence indicates germinal centers. c, Images of p-LN sections harvested from a treated mouse at day 23, two days after BL-PDT. d, Images of the regions marked in b and c. Arrows indicate damaged blood vessels. e, Integrated melan-A fluorescence magnitudes of the untreated (U) and treated (T) p-LNs. f, Inguinal LN sections harvested from the untreated mouse. g, Inguinal LN sections harvested from the treated mouse. h, The regions marked in f and g. Scale bars, 500 µm. Error bars, s.d. **, Kruskal-Wallis test p <0.01.
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Figure 6: Treatment of lymph-node metastases. a, BL image of a C57/BL6 mouse with B16F10 tumor in the footpad (day 21), after injection of Luc-QD and CTZ into the foot. b, IHC images of p-LN sections harvested from an untreated mouse at day 23, stained for blood vessels (CD31), lymphatics (VEGFR3/CD31), melanoma cells (melan-A), and apoptosis (caspase-3). Strong green autofluorescence indicates germinal centers. c, Images of p-LN sections harvested from a treated mouse at day 23, two days after BL-PDT. d, Images of the regions marked in b and c. Arrows indicate damaged blood vessels. e, Integrated melan-A fluorescence magnitudes of the untreated (U) and treated (T) p-LNs. f, Inguinal LN sections harvested from the untreated mouse. g, Inguinal LN sections harvested from the treated mouse. h, The regions marked in f and g. Scale bars, 500 µm. Error bars, s.d. **, Kruskal-Wallis test p <0.01.

Mentions: To test BL-PDT for orthotopic melanoma, 1.5x106 B16F10 cells were injected into the footpad of wild-type C57BL/6 mice. BL emission from the p-LN was confirmed, although it was less bright than in the CT-26 bearing nude mice due to the presence of hair and high melanin in the tumor cells (Fig. 6a). BL-PDT was performed 21 days after implantation. At day 23, the popliteal and inguinal LNs were harvested from both treated and untreated groups, sectioned, and examined by immunohistochemistry. The untreated p-LN had a considerable number of tumor cells, indicated by melan-A surface antigen, scattered in multiple groups throughout the paracortex and medulla (Fig. 6b). The treated p-LN exhibited several distinct consequences of BL-PDT including the remarkable increase of apoptotic cell deaths throughout the cortex and paracortex (Fig. 6c). In these regions, the tumor vessels and lymphatic sinuses appear to have been severely disintegrated (Fig. 6d). The amount of melan-A signal decreased to a half after the treatment in the p-LNs (Fig. 6e). The effects of BL-PDT were not limited to sentinel LN. Tumor cells were also found at inguinal LNs in the untreated mouse, particularly along the lymphatic vessels (Fig. 6f). At inguinal LN in the treated mouse (Fig. 6g), considerable apoptosis signals were measured in the paracortex and medulla. Notably, the number of melanoma cells at inguinal LN was significantly reduced in the treated compared to untreated mice (Fig. 6h).


Bioluminescence-activated deep-tissue photodynamic therapy of cancer.

Kim YR, Kim S, Choi JW, Choi SY, Lee SH, Kim H, Hahn SK, Koh GY, Yun SH - Theranostics (2015)

Treatment of lymph-node metastases. a, BL image of a C57/BL6 mouse with B16F10 tumor in the footpad (day 21), after injection of Luc-QD and CTZ into the foot. b, IHC images of p-LN sections harvested from an untreated mouse at day 23, stained for blood vessels (CD31), lymphatics (VEGFR3/CD31), melanoma cells (melan-A), and apoptosis (caspase-3). Strong green autofluorescence indicates germinal centers. c, Images of p-LN sections harvested from a treated mouse at day 23, two days after BL-PDT. d, Images of the regions marked in b and c. Arrows indicate damaged blood vessels. e, Integrated melan-A fluorescence magnitudes of the untreated (U) and treated (T) p-LNs. f, Inguinal LN sections harvested from the untreated mouse. g, Inguinal LN sections harvested from the treated mouse. h, The regions marked in f and g. Scale bars, 500 µm. Error bars, s.d. **, Kruskal-Wallis test p <0.01.
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Related In: Results  -  Collection

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Figure 6: Treatment of lymph-node metastases. a, BL image of a C57/BL6 mouse with B16F10 tumor in the footpad (day 21), after injection of Luc-QD and CTZ into the foot. b, IHC images of p-LN sections harvested from an untreated mouse at day 23, stained for blood vessels (CD31), lymphatics (VEGFR3/CD31), melanoma cells (melan-A), and apoptosis (caspase-3). Strong green autofluorescence indicates germinal centers. c, Images of p-LN sections harvested from a treated mouse at day 23, two days after BL-PDT. d, Images of the regions marked in b and c. Arrows indicate damaged blood vessels. e, Integrated melan-A fluorescence magnitudes of the untreated (U) and treated (T) p-LNs. f, Inguinal LN sections harvested from the untreated mouse. g, Inguinal LN sections harvested from the treated mouse. h, The regions marked in f and g. Scale bars, 500 µm. Error bars, s.d. **, Kruskal-Wallis test p <0.01.
Mentions: To test BL-PDT for orthotopic melanoma, 1.5x106 B16F10 cells were injected into the footpad of wild-type C57BL/6 mice. BL emission from the p-LN was confirmed, although it was less bright than in the CT-26 bearing nude mice due to the presence of hair and high melanin in the tumor cells (Fig. 6a). BL-PDT was performed 21 days after implantation. At day 23, the popliteal and inguinal LNs were harvested from both treated and untreated groups, sectioned, and examined by immunohistochemistry. The untreated p-LN had a considerable number of tumor cells, indicated by melan-A surface antigen, scattered in multiple groups throughout the paracortex and medulla (Fig. 6b). The treated p-LN exhibited several distinct consequences of BL-PDT including the remarkable increase of apoptotic cell deaths throughout the cortex and paracortex (Fig. 6c). In these regions, the tumor vessels and lymphatic sinuses appear to have been severely disintegrated (Fig. 6d). The amount of melan-A signal decreased to a half after the treatment in the p-LNs (Fig. 6e). The effects of BL-PDT were not limited to sentinel LN. Tumor cells were also found at inguinal LNs in the untreated mouse, particularly along the lymphatic vessels (Fig. 6f). At inguinal LN in the treated mouse (Fig. 6g), considerable apoptosis signals were measured in the paracortex and medulla. Notably, the number of melanoma cells at inguinal LN was significantly reduced in the treated compared to untreated mice (Fig. 6h).

Bottom Line: Owing to the shallow penetration of light in tissues, however, the clinical applications of light-activated therapies have been limited.For monolayer cell culture in vitro incubated with Chlorin e6, BRET energy of about 1 nJ per cell generated as strong cytotoxicity as red laser light irradiation at 2.2 mW/cm(2) for 180 s.Our results show the promising potential of novel bioluminescence-activated PDT.

View Article: PubMed Central - PubMed

Affiliation: 1. Graduate School of Nanoscience and Technology (WCU), Korea Advanced Institute of Science and Technology, 291 Daehak-Ro, Yusong-Gu, Daejon 305-701, Korea ; 2. Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, 291 Daehak-Ro, Yusong-Gu, Daejon 305-701, Korea ; 3. Department of Oncology, Asan Medical Center, Univ. Ulsan College of Medicine, Seoul , Korea.

ABSTRACT
Optical energy can trigger a variety of photochemical processes useful for therapies. Owing to the shallow penetration of light in tissues, however, the clinical applications of light-activated therapies have been limited. Bioluminescence resonant energy transfer (BRET) may provide a new way of inducing photochemical activation. Here, we show that efficient bioluminescence energy-induced photodynamic therapy (PDT) of macroscopic tumors and metastases in deep tissue. For monolayer cell culture in vitro incubated with Chlorin e6, BRET energy of about 1 nJ per cell generated as strong cytotoxicity as red laser light irradiation at 2.2 mW/cm(2) for 180 s. Regional delivery of bioluminescence agents via draining lymphatic vessels killed tumor cells spread to the sentinel and secondary lymph nodes, reduced distant metastases in the lung and improved animal survival. Our results show the promising potential of novel bioluminescence-activated PDT.

No MeSH data available.


Related in: MedlinePlus