Evaluation of oxygen dependence on in vitro and in vivo cytotoxicity of photoimmunotherapy using IR-700–antibody conjugates

Kishimoto, Shun; Bernardo, Marcelino; Saito, Keita; Koyasu, Sho; Mitchell, James B.; Choyke, Peter L.; Krishna, Murali C.

doi:10.1016/j.freeradbiomed.2015.03.038

Cited by 49 publications

(48 citation statements)

References 49 publications

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“…5,12 Unlike conventional plasma membrane-targeted photosensitizers, an mAb-IR700 is sufficiently far away from the plasma membrane and does not induce oxidative damage because of the very short half-life of singlet oxygen. 12,34 There is a report that free radical scavengers reduced the effect of NIR-PIT, 35 although this has not been generally accepted. 5,11 The molecular mechanism causing such minute plasma membrane damage in NIR-PIT remains unclear.…”

Section: Discussionmentioning

confidence: 99%

Changes in plasma membrane damage inducing cell death after treatment with near‐infrared photoimmunotherapy

et al. 2018

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Near‐infrared photoimmunotherapy (NIR‐PIT) is a new cancer phototherapy modality using an antibody conjugated to a photosensitizer, IRDye700DX. When the conjugate binds to the plasma membrane and is exposed to NIR light, NIR‐PIT‐treated cells undergo swelling, and target‐selective necrotic/immunogenic cell death is induced. However, the cytotoxic mechanism of NIR‐PIT has not been elucidated. In order to understand the mechanism, it is important to elucidate how the damage to the plasma membrane induced by NIR light irradiation changes over time. Thus, in the present study, we investigated the changes in plasma membrane permeability using ions and molecules of various sizes. Na+ flowed into cells immediately after NIR light irradiation, even when the function of transporters or channels was blocked. Subsequently, fluorescent molecules larger than Na+ entered the cells, but the damage was not large enough for dextran to pass through at early time points. To assess these phenomena quantitatively, membrane permeability was estimated using radiolabeled ions and molecules: 111InCl3, 111In‐DTPA, and 3H‐H2O, and comparable results were obtained. Although minute plasma membrane perforations usually do not induce cell death, our results suggest that the minute damage induced by NIR‐PIT was irreversibly extended with time. In conclusion, minute plasma membrane damage is a trigger for the increase in plasma membrane permeability, cell swelling, and necrotic/immunogenic cell death in NIR‐PIT. Our findings provide new insight into the cytotoxic mechanism of NIR‐PIT.

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Section: Discussionmentioning

confidence: 99%

Changes in plasma membrane damage inducing cell death after treatment with near‐infrared photoimmunotherapy

et al. 2018

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“…BT142 seemed to have the characteristics of apoptosis and necrosis within 60 m.[21,22] It is consistent with the reported result that photodamage of A431-cells by IR700 induced both apoptosis and necrosis. [23] At 2 h after photoactivation, vesicles in BT142 cells diffused in the same degree as U87MG cells and the fragmentation of the nucleus detected by EthD-1 was not observed, suggesting necrotic cell death. These results indicate that BT142 had characteristics of necrotic and apoptotic cell death in an early stage of cell damage and progressed in necrotic cell death at a later stage.…”

Section: Discussionmentioning

confidence: 99%

Quantitative evaluation of malignant gliomas damage induced by photoactivation of IR700 dye

Sakuma

Kita

Higuchi

2016

Science and Technology of Advanced Materials

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The processes involved in malignant gliomas damage were quantitatively evaluated by microscopy. The near-infrared fluorescent dye IR700 that is conjugated to an anti-CD133 antibody (IR700-CD133) specifically targets malignant gliomas (U87MG) and stem cells (BT142) and is endocytosed into the cells. The gliomas are then photodamaged by the release of reactive oxygen species (ROS) and the heat induced by illumination of IR700 by a red laser, and the motility of the vesicles within these cells is altered as a result of cellular damage. To investigate these changes in motility, we developed a new method that measures fluctuations in the intensity of phase-contrast images obtained from small areas within cells. The intensity fluctuation in U87MG cells gradually decreased as cell damage progressed, whereas the fluctuation in BT142 cells increased. The endocytosed IR700 dye was co-localized in acidic organelles such as endosomes and lysosomes. The pH in U87MG cells, as monitored by a pH indicator, was decreased and then gradually increased by the illumination of IR700, while the pH in BT142 cells increased monotonically. In these experiments, the processes of cell damage were quantitatively evaluated according to the motility of vesicles and changes in pH.

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“…Photoactivation of IR700 was recently developed to specifically damage cancer cells in vitro and in vivo (Harada et al, 2015; Nagaya et al, 2016). Previous research has shown that photoactivation induces cellular damage by producing ROS and heat (Mitsunaga et al, 2011; Kishimoto et al, 2015). To specify the effect of photoactivation, we applied a motility assay of kinesin and dynein to microtubules (Fig.…”

Section: Discussionmentioning

confidence: 99%

Quantitative Detection of Cell Activity by Measuring the Fluctuation of Intracellular Motility

Sakuma

Kondo

Higuchi

2019

Preprint

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13The measurement of cell activity changes during damage is important to 14 understand the process of cell death and evaluate the effect of medicines. To 15 evaluate cell activity generally, we extended the method of intensity fluctuation in 16 which intensity change in the pixel induced by the movement of organelles was 17 calculated. Cancer, endothelial and iPS cells were damaged by reactive oxygen 18 species (ROS) generated by a fluorescent dye (IR700), hydrogen peroxide, and 19 ultraviolet light. The intensity fluctuation in damaged cells gradually decreased 20 independent of the kind of cell, indicating that the decrease in the fluctuation is a 21 general phenomenon in damaged cells. The rupture of vesicles and 22 mitochondria in the cells were observed upon ROS production. The motility of 23 purified kinesin and dynein which transport vesicles and organelles was inhibited 24 by ROS. These suggest that ROS and cytotoxic molecules spreading from 25 ruptured organelles contribute to the reduction in cell activity which brings about 26 the decrease in the motility and intensity fluctuation of organelles driven by 27 kinesin and dynein.28 Liao et al., 2017). The motility of mitochondria gradually decreased just after the 65 addition of hydrogen peroxide, and the decrease in the motility reached 50% 66 within 10 minutes (Debattisti et al., 2017). These results suggested that the 67 effect on the motility of organelles in damaged cells occurred earlier than the 68 changes in cell morphology and motility. Thus, the method of detecting 69 intracellular motility under a conventional microscope would overcome the 70 disadvantages of previous fluorescence, morphological, and cell motility 71 methods. However, little is understood about the generality and mechanism of 72 motility reduction. 73In this study, various cells were damaged by several methods, including 74 photoactivation of IR700, irradiation with ultraviolet (UV) light and treatment with 75 hydrogen peroxide (H 2 O 2 ), and the change in the motility of organelles was 76 quantitatively evaluated by the developed intensity fluctuation method (IFM). We 77 found that all cells showed a decrease in motility caused by damage. To 78 elucidate the mechanisms of the decrease in motility in damaged cells, we 79 measured the reduction in the motile speeds of single vesicles in cells and in 80 purified kinesin and dynein in vitro assays. These measurements revealed that 81 the decrease detected by IMF resulted in the reduction in the organelle motility 82 driven by motor proteins. Since the transport system commonly exists in animal 83 cells, the decrease in the motility or transport of organelles would be a universal 84 phenomenon in various kinds of cells. 85 5 Results 86 87 1 Decrement of motility of organelles in damaged cells 88 Changes in the motility of organelles in damaged cells were evaluated 89 by the intensity fluctuation method (IFM) in various cell types. Six kinds of cells, 90 four cancer cell lines, one endothelial cell line, and one iPS cell line...

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Evaluation of oxygen dependence on in vitro and in vivo cytotoxicity of photoimmunotherapy using IR-700–antibody conjugates

Cited by 49 publications

References 49 publications

Changes in plasma membrane damage inducing cell death after treatment with near‐infrared photoimmunotherapy

Changes in plasma membrane damage inducing cell death after treatment with near‐infrared photoimmunotherapy

Quantitative evaluation of malignant gliomas damage induced by photoactivation of IR700 dye

Quantitative Detection of Cell Activity by Measuring the Fluctuation of Intracellular Motility

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