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(A) Representative normalized average IR700 fluorescence intensity (upper) and GFP intensity (lower) of tumor surface and deep tumor during and after PIT treatment compared with control mouse. (B) Left: IR700 fluorescence recovery value of tumor surface and deep tumor. Deep tumor showed significantly higher fluorescence recovery intensity value than that of the tumor surface (n = 5). Right: GFP intensity of tumor surface and deep tumor. The tumor surface showed a significantly larger GFP fluorescence decrease than that of the deep tumor (n = 5). (C) Histological specimens of A431 tumors from the PIT-treated group. The image on the right side is the zoomed-in image from the blue square in the left image. The yellow dashed circles indicate undamaged cells. Overall, there was less damage compared to the two-channel fiber bundle results. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
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![Fig. 2. (A) Changes in fluorescence [ΔF/F (%), ordinate] of the tumor...](profile/Qinggong-Tang/publication/317407255/figure/fig2/AS:567694699098112@1512360371248/A-Changes-in-fluorescence-DF-F-ordinate-of-the-tumor-surface-and-deep-tumor_Q320.jpg)
Photoimmunotherapy (PIT) is an emerging low side effect cancer therapy based on a monoclonal antibody (mAb) conjugated with a near-infrared (NIR) phthalocyanine dye IRDye 700DX. IR700 is fluorescent, can be used as an imaging agent, and also is phototoxic. It induces rapid cell death after exposure to NIR light. PIT induces highly selective cancer...
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... Fig. 5A, the normalized IR700 fluorescence intensity averaged from the white and blue squares showed a greater drop in the tumor surface and greater recovery in deep tumor, while there is no obvious change in controls. The recovery of IR700 fluorescence intensity in deep tumor is significantly higher than that of the tumor surface and > 10% ...
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... intensity averaged from the white and blue squares showed a greater drop in the tumor surface and greater recovery in deep tumor, while there is no obvious change in controls. The recovery of IR700 fluorescence intensity in deep tumor is significantly higher than that of the tumor surface and > 10% higher than the initial value of deep tumor (Fig. 5B). The normalized GFP fluorescence intensity showed a larger drop in the tumor region (both tumor surface and deep tumor) treated with PIT compared to the decrease in controls (lower image and chart in Fig. 5A). The decrease of GFP fluorescence intensity in the tumor surface is significantly greater than that of the deep tumor (Fig. ...
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... intensity in deep tumor is significantly higher than that of the tumor surface and > 10% higher than the initial value of deep tumor (Fig. 5B). The normalized GFP fluorescence intensity showed a larger drop in the tumor region (both tumor surface and deep tumor) treated with PIT compared to the decrease in controls (lower image and chart in Fig. 5A). The decrease of GFP fluorescence intensity in the tumor surface is significantly greater than that of the deep tumor (Fig. ...
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... deep tumor (Fig. 5B). The normalized GFP fluorescence intensity showed a larger drop in the tumor region (both tumor surface and deep tumor) treated with PIT compared to the decrease in controls (lower image and chart in Fig. 5A). The decrease of GFP fluorescence intensity in the tumor surface is significantly greater than that of the deep tumor (Fig. ...
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... H & E stained sections, more necrotic cells were seen in the tumor surface than in the deep tumor (Fig. 5C) and there are cells remaining undamaged (yellow dashed area in Fig. 5C). There is overall less da- mage compared to the two-channel fiber bundle results. In the control tumor, no obvious cell necrosis was observed in either tumor surface or the deep tumor. The micro-prism track can be clearly seen ~1.4-1.7 mm below the surface ...
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... H & E stained sections, more necrotic cells were seen in the tumor surface than in the deep tumor (Fig. 5C) and there are cells remaining undamaged (yellow dashed area in Fig. 5C). There is overall less da- mage compared to the two-channel fiber bundle results. In the control tumor, no obvious cell necrosis was observed in either tumor surface or the deep tumor. The micro-prism track can be clearly seen ~1.4-1.7 mm below the surface (Supplementary Fig. ...
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... in- tensity in the tumor surface was significantly higher than that of control tumors and ~1.6-fold greater than initial values (Fig. 6C). The decrease of GFP fluorescence intensity in the tumor surface was significantly greater than that of the control tumors (Fig. 6C) and also greater than that of the tumor surface viewed using microprism (Fig. ...
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As a novel low-side-effect cancer therapy, photo-immunotherapy (PIT) is based on conjugating monoclonal antibody (mAb) with a near-infrared (NIR) phthalocyanine dye IRDye700DX (IR 700). IR700 is not only fluorescent to be used as an imaging agent, but also phototoxic. When illuminating with NIR light, PIT can induce highly-selective cancer cell dea...
Citations
... Kobayashi et al. found that the NIR phthalocyanine dye, IR700, conjugated to mAbs (mAb-IR700), along with the use of NIR light irradiation, enhanced the targetselective anti-tumor performance with immune activation. [63][64][65][66][67][68][69][70][71][72][73][74][75][76][77][78][79] As shown in Figure 2a and b, the tumor volume was significantly reduced after NIR-PIT with mAb-IR700 treatment compared to the untreated control, while survival was prolonged in mice. In addition, dendritic cell (DC) maturation was confirmed using CD80 and CD86 markers after NIR-PIT-mediated cancer cell death (Figure 2c). ...
Light-triggered phototherapies, such as photodynamic therapy (PDT) and photothermal therapy (PTT), have shown strong therapeutic efficacy with minimal invasiveness and systemic toxicity, and offer opportunities for tumor-specific therapies. Phototherapies not...
... The QD800-fluorescence-signal besides the vascular was predominantly and significantly more increased in the NIR-PIT (100 J/cm 2 ) group (n = 6, p = 0.00000108 < 0.05, vs. vessels, the larger the pan-IR700 distribution, thereby increasing the space around to the vessels, as consistent to previously reported. 48,49 Evaluation of the SUPR effect after NIR-PIT using NIR-PLGA microparticles Conventional EPR and previous reported SUPR effects with NIR-PIT have been thought to be exerted by nanoparticles sized 10∼200 nm. However, the 3Dcleared tissue images suggested that dilated tumour vessels were stretched around inside the NIR-PITtreated tumours (Fig. 3e and f). ...
Background:
Near-infrared photoimmunotherapy (NIR-PIT) is a promising cancer therapy combining NIR-light irradiation with an antibody and IR700DX, a light-sensitive substance, to destroy tumours. However, homogeneous irradiation is difficult because the light varies depending on the distance and tissue environment. Therefore, markers that indicate sufficient irradiation are necessary. Nanoparticles sized 10∼200 nm show enhanced permeation and retention within tumours, which is further enhanced via NIR-PIT (super enhanced permeability and retention, SUPR). We aimed to monitor the effectiveness of NIR-PIT by measuring SUPR.
Methods:
A xenograft mouse tumour model was established by inoculating human cancer cells in both buttocks of Balb/C-nu/nu mice, and NIR-PIT was performed on only one side. To evaluate SUPR, fluorescent signal examination was performed using QD800-fluorescent nanoparticles and NIR-fluorescent poly (d,l-lactide-co-glycolic acid) (NIR-PLGA) microparticles. Harmonic signals were evaluated using micro-bubbles of the contrast agent Sonazoid and contrast-enhanced ultrasound (CEUS) imaging. The correlation between SUPR immediately after treatment and NIR-PIT effectiveness on the day after treatment was evaluated.
Findings:
QD800 fluorescent signals persisted only in the treated tumours, and the intensity of remaining signals showed high positive correlation with the therapeutic effect. NIR-PLGA fluorescent signals and Sonazoid-derived harmonic signals remained for a longer time in the treated tumours than in the controls, and the kE value of the two-compartment model correlated with NIR-PIT effectiveness.
Interpretation:
SUPR measurement using Sonazoid and CEUS imaging could be easily adapted for clinical use as a therapeutic image-based biomarker for monitoring and confirming of NIR-PIT efficacy.
Funding:
This research was supported by ARIM JAPAN of MEXT, the Program for Developing Next-generation Researchers (Japan Science and Technology Agency), KAKEN (18K15923, 21K07217) (JSPS), CREST (JPMJCR19H2, JST), and FOREST-Souhatsu (JST). Mochida Memorial Foundation for Medical and Pharmaceutical Research; Takeda Science Foundation; The Japan Health Foundation; and Princess Takamatsu Cancer Research Fund. Funders only provided financial support and had no role in the study design, data collection, data analysis, interpretation, and writing of the report.
... 24,26 Other than special drugs, SUPR effects also permit enhanced delivery for other antibodies and APCs, whose intertumoral distribution is improved by enlarged leakage into the tumor bed after NIR-PIT. [27][28][29] A study utilizing FDA approved daunorubicin (DaunoXome) encapsulated with liposome and nanoparticle paclitaxel (nab-paclitaxel; Abraxane) bound with albumin-in mouse xenograft models of cancer, demonstrated that NIR-PIT cooperated with either drug showed greatest advantage. 30,31 Furthermore, uncaging reactions induced by NIR light could transmit bioactive compounds encapsuled in particles to any favorable site of the body, illustrating a remarkable chemical progress and prospect of ongoing study. ...
Worldwide, the incidence rate of breast cancer is the highest in women. Approximately 2.3 million people were newly diagnosed and 0.685 million were dead of breast cancer in 2020, which continues to grow. Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype with a higher risk of recurrence and metastasis, but disappointly, there are no effective and specific therapies clinically, especially for patients presenting with metastatic diseases. Therefore, it is urgent to develop a new type of cancer therapy for survival improvisation and adverse effects alleviation of breast cancers. Near-infrared photoimmunotherapy (NIR-PIT) is a newly developed, photochemistry-based cancer therapy. It was drive by an antibody-photoabsorber conjugate (APC) which is triggered by near-infrared light. The key part of APC is a cancer-targeting monoclonal antibody (mAb) that can bind to receptors or antigens on the surface of tumor cells. Because of this targeted conjugate accumulation, subsequent deployment of focal NIR-light results in functional damage on the targeted cell membranes without harming the immediately adjacent receptor-negative cells and evokes a kind of photochemical, speedy, and highly specific immunogenic cell death (ICD) of cancer cells with corresponding antigens. Subsequently, immature dendritic cells adjacent to dying cancer cells will become mature, further inducing a host-oriented anti-cancer immune response, complicatedly and comprehensively. Currently, NIR-PIT has progressed into phase 3 clinical trial for recurrent head and neck cancer. And preclinical studies have illustrated strong therapeutic efficacy of NIR-PIT targeting various molecular receptors overexpressed in breast cancer cells, including EGFR, HER2, CD44c, CD206, ICAM-1 and FAP-α. Thereby, NIR-PIT is in early trials, but appears to be a promising breast cancer therapy and moving into the future. Here, we present the specific advantages and discuss the most recent preclinical studies against several transmembrane proteins of NIR-PIT in breast cancers.
... This imaging system shows a significant difference in treated tumors compared with untreated tumors [95]. Moreover, the micro-distribution of the NIR-PIT agent from the tumor surface to the deep tumor during and after treatment and its therapeutic effects is monitored using a two-channel fluorescence fiber imaging system and two-photon microscopy with and without a microprism [96]. ...
Near infrared photoimmunotherapy (NIR-PIT) is a newly developed molecular targeted cancer treatment, which selectively kills cancer cells or immune-regulatory cells and induces therapeutic host immune responses by administrating a cancer targeting moiety conjugated with IRdye700. The local exposure to near-infrared (NIR) light causes a photo-induced ligand release reaction, which causes damage to the target cell, resulting in immunogenic cell death (ICD) with little or no side effect to the surrounding normal cells. Moreover, NIR-PIT can generate an immune response in distant metastases and inhibit further cancer attack by combing cancer cells targeting NIR-PIT and immune regulatory cells targeting NIR-PIT or other cancer treatment modalities. Several recent improvements in NIR-PIT have been explored such as catheter-driven NIR light delivery, real-time monitoring of cancer, and the development of new target molecule, leading to NIR-PIT being considered as a promising cancer therapy. In this review, we discuss the progress of NIR-PIT, their mechanism and design strategies for cancer treatment. Furthermore, the overall possible targeting molecules for NIR-PIT with their application for cancer treatment are briefly summarised.
... After tumor transmission, 48 other antibodies or APCs can be promoted to the tumor bed. 50 The tumor-bearing mouse model showed that multiple illuminations after a single dose of APC could significantly inhibit tumor growth. 50 This was due to the enhanced effect induced by initial illumination so that APC existing in blood could enter larger tumor areas and deep tumor tissues that were difficult to reach before PIT, and its distribution was more uniform. ...
... 50 The tumor-bearing mouse model showed that multiple illuminations after a single dose of APC could significantly inhibit tumor growth. 50 This was due to the enhanced effect induced by initial illumination so that APC existing in blood could enter larger tumor areas and deep tumor tissues that were difficult to reach before PIT, and its distribution was more uniform. 51 ...
... Solid tumors composed of phenotypic and functionally heterogeneous malignant cells can interfere with tumor treatment, especially the effectiveness of targeted molecular therapies, including PIT, and ultimately lead to tumor recurrence. 50 Compared with DX or PIT alone, the tumor volume of the mice treated with PIT and DX was significantly reduced, and the survival time was significantly prolonged. In another study, 49 glypican-3 (GPC3)targeted PIT combined with Abraxane showed superior efficacy in treating hepatocellular carcinoma. ...
In recent years, the incidence of cancer has been increasing worldwide. Conventional cancer treatments include surgery, chemotherapy, and radiation, which mostly kill tumor cells at the expense of normal and immune cells. Although immunotherapy is an accurate, rapid, efficient tumor immune treatment, it causes serious adverse reactions, such as cytokine release syndrome (CRS) and neurotoxicity. Therefore, there is an urgent need to develop an effective and nontoxic procedure for immunotherapy. The clinical combination of phototherapy and immunoadjuvant therapy can induce immunogenic cell death and enhance antigen presentation synergy. It also causes a systemic antitumor immune response to manage residual tumors and distant metastases. Photoimmunotherapy (PIT) is a tumor treatment combining phototherapy with immunotherapy based on injecting a conjugate photosensitizer (IR700) and a monoclonal antibody (mAb) to target an expressed antigen on the tumor surface. This combination can enhance the immune response ability, thus having a good effect on the treatment of residual tumor and metastatic cancer. In this review, we summarize the recent progress in photoimmunotherapy, including photoimmunoconjugate (PIC), the activation mechanism of immunogenic cell death (ICD), the combination therapy model, opportunities and prospects. Specifically, we aim to provide a promising clinical therapy for solid tumor clinical transformation.
... It not only helps researchers to understand the molecular characteristics of the disease but also facilitates identification of the cancerous tissues. One of the unique advantages of photoimmuno- conjugates is the ability to leverage both the fluorescence capability of the photosensitizers and the selectivity of the antibody to specifically label cancer cells for fluorescence visualization [46]. We tested if UIC2-BPD could be used for BPD fluorescence imaging of ABCB1-overexpressing TNBC cells. ...
Accurate detection of ATP-binding cassette drug transporter ABCB1 expression is imperative for precise identification of drug-resistant tumors. Existing detection methods fail to provide the necessary molecular details regarding the functional state of the transporter. Photoimmunoconjugates are a unique class of antibody–dye conjugates for molecular diagnosis and therapeutic treatment. However, conjugating hydrophobic photosensitizers to hydrophilic antibodies is quite challenging. Here, we devise a photoimmunoconjugate that combines a clinically approved benzoporphyrin derivative (BPD) photosensitizer and the conformational-sensitive UIC2 monoclonal antibody to target functionally active human ABCB1 (i.e., ABCB1 in the inward-open conformation). We show that PEGylation of UIC2 enhances the BPD conjugation efficiency and reduces the amount of non-covalently conjugated BPD molecules by 17%. Size exclusion chromatography effectively separates the different molecular weight species found in the UIC2–BPD sample. The binding of UIC2–BPD to ABCB1 was demonstrated in lipidic nanodiscs and ABCB1-overexpressing triple negative breast cancer (TNBC) cells. UIC2–BPD was found to retain the conformation sensitivity of UIC2, as the addition of ABCB1 modulators increases the antibody reactivity in vitro . Thus, the inherent fluorescence capability of BPD can be used to label ABCB1-overexpressing TNBC cells using UIC2–BPD. Our findings provide insight into conjugation of hydrophobic photosensitizers to conformation-sensitive antibodies to target proteins expressed on the surface of cancer cells.
... Before imaging, the skin above the tumor was removed and a window chamber was used to fix the tumor for minimizing the imaging artifact from animal movement as previous reported [56,62]. Near infrared LED used in our experiments has a wavelength spectrum of 680 -700 nm (Tech-LED, Marubeni America Corp., Santa Clara, CA, USA) [63,64]. LED was turned on for ∼30 min with a dose of ∼50 J/cm 2 to irradiate the tumor [64]. ...
... Near infrared LED used in our experiments has a wavelength spectrum of 680 -700 nm (Tech-LED, Marubeni America Corp., Santa Clara, CA, USA) [63,64]. LED was turned on for ∼30 min with a dose of ∼50 J/cm 2 to irradiate the tumor [64]. To record the alterations of tumor vasculature during and after NIR-PIT caused by LED, the data acquisition protocol is as follows: An image of tumor was taken every 10 min for the first 30 min before LED was turned on. ...
Near-infrared (NIR) photoimmunotherapy (NIR-PIT) is an emerging cancer therapy based on a monoclonal antibody and phthalocyanine dye conjugate. Direct tumor necrosis and immunogenic cell death occur during NIR irradiation. However, the alteration of tumor blood vessels and blood volume inside the blood vessels induced by the NIR-PIT process is still unknown. In our study, a speckle variance (SV) algorithm combined with optical coherence tomography (OCT) technology was applied to monitor the change of blood vessels and the alterations of the blood volume inside the blood vessels during and after NIR-PIT treatment. Vascular density and the measurable diameter of the lumen in the blood vessel (the diameter of the region filled with blood) were extracted for quantitively uncovering the alterations of blood vessels and blood volume induced by NIR-PIT treatment. The results indicate that both the density and the diameter of the lumen in the blood vessels decrease during the NIR-PIT process, while histological results indicated the blood vessels were dilated. The increase of permeability of blood vessels could lead to the increase of the blood pool volume within the tumor (shown in histology) and results in the decrease of free-moving red blood cells inside the blood vessels (shown in SV-OCT).
... Therefore, a combination therapy using NIR-PIT followed by a nano-sized anticancer agent could be more effective than either therapy alone [4,18]. SUPR effects also allow for enhanced delivery for other antibodies and APCs, whose intra-tumoral distribution is enhanced by increased leakage into the tumor bed after NIR-PIT [20][21][22]. ...
Background:
Cancers of the gastrointestinal (GI) tract are the common leading cause of cancer-related death in the world. Recent advances in cancer therapies such as intensive multidrug chemotherapy and molecular targeted treatment have improved therapeutic efficacy; however, the outcomes are not satisfied. Moreover, these therapies also cause severe side effects. New type of cancer therapies is urgently needed to improve the outcomes and to reduce side effects of GI tract cancers.
Summary:
This account is a comprehensive review article on the newly developed, photochemistry-based cancer therapy named as near-infrared photoimmunotherapy (NIR-PIT). NIR-PIT is a highly selective tumor treatment that employs an antibody-photoabsorber conjugate, which is activated by near-infrared light. A world-wide phase 3 clinical trial of NIR-PIT against recurrent head and neck cancer patients is currently underway. NIR-PIT differs from conventional cancer therapies such as surgery, chemotherapy, and radiation in its selectivity for killing cancer cells and cells treated with NIR-PIT leading to immunogenic cell death. Preclinical research in animals with combining cancer-targeting NIR-PIT and other cancer immunotherapies could lead to responses not only in local tumor but also in distant metastases. NIR-PIT also leads to an immediate and dramatic increase in vascular permeability after therapy. From these aspects, NIR-PIT appears to be a promising new form of cancer therapy. NIR-PIT could be readily translated into clinical use for virtually any cancers in the near future provided suitable humanized antibodies are available. Here, we describe the specific advantages and applications of NIR-PIT in the GI tract. Key Messages: We believe that NIR-PIT with NIR excitation light, which can be delivered via a fiber optic diffuser through endoscopes, is a promising method for a new treatment of GI cancers.
... 21 This is the basis for the SUPR effect, which peaks within 6 hours of treatment and returns to near baseline by 24 hours. 22 The SUPR effects allow residual APC in the circulation to penetrate further into tumor tissue and enhanced delivery of nanodrugs after therapy. This effect is profound resulting in up to 24-fold increase delivery of nanodrugs into the tumor compared to nontreated tumors. ...
... This effect is profound resulting in up to 24-fold increase delivery of nanodrugs into the tumor compared to nontreated tumors. 4,7,8,22 Therefore, the SUPR effect has favorable implications for repeated light exposures and fractionated administration of combination therapies. Here we show that it can also be useful for monitoring the selective tumor uptake of the albumin-bound fluorescent dye, ICG. ...
Background
Near-infrared photoimmunotherapy (NIR-PIT) is a cancer therapy that causes an increase in tumor perfusion, a phenomenon termed the super-enhanced permeability and retention effect. Currently, in vivo treatment efficacy of NIR-PIT is observable days after treatment, but monitoring would be improved by more acute detection of intratumor change. Fluorescence imaging may detect increased tumor perfusion immediately after treatment.
Methods
In the first experiment, athymic nude mouse models bearing unilateral subcutaneous flank tumors were treated with either NIR-PIT or laser therapy only. In the second experiment, mice bearing bilateral flank tumors were treated with NIR-PIT only on the left-sided tumor. In both groups, immediately after treatment, indocyanine green was injected at different doses intravenously, and mice were monitored with the Shimadzu LIGHTVISION fluorescence imaging system for 1 hour.
Results
Tumor-to-background ratio of fluorescence intensity increased over the 60 minutes of monitoring in treated mice but did not vary significantly in control mice. Tumor-to-background ratio was highest in the 1 mg kg ⁻¹ and 0.3 mg kg ⁻¹ doses. In mice with bilateral tumors, tumor-to-untreated tumor ratio increased similarly.
Conclusions
Acute changes in tumor perfusion after NIR-PIT can be detected by real-time fluorescence imaging.
... The peripheral blood velocity drop is due to the increase of blood pool volume within the treated tumor and implies a long drug circulation time, which can explain the hemodynamic basis of the SUPR effect. Tang et al. examined the microdistribution of APCs during and after NIR-PIT using a minimally invasive twochannel fluorescence fiber imaging system and a high-resolution two-photon microscope with a microprism [120]. The IR700 fluorescence intensity showed a quick drop both at the tumor surface and deep within the tumor during the NIR exposure, recovering quickly thereafter. ...
... The normalized IR700 fluorescence intensity shows a greater drop in the tumor surface (PIT-s) and a higher recovery in deep tumor (PIT-d), although there was only a gradual decrease in controls (control-s and control-d). Adapted from Ref.[120]. ...
To date, the delivery of nanosized therapeutic agents to cancers largely relies on the enhanced permeability and retention (EPR) effects that are caused by the leaky nature of cancer vasculature. Whereas leaky vessels are often found in mouse xenografts, nanosized agents have demonstrated limited success in humans due to the relatively small magnitude of the EPR effect in naturally occurring cancers. To achieve the superior delivery of nanosized agents, alternate methods of increasing permeability and retention are needed. Near-infrared photoimmunotherapy (NIR-PIT) is a recently reported therapy that relies on an antibody-photon absorber conjugate that binds to tumors and then is activated by light. NIR-PIT causes an increase in nanodrug delivery by up to 24-fold compared to untreated tumors in which only the EPR effect is present. This effect, termed super-EPR (SUPR), can enhance the delivery of a wide variety of nanosized agents, including nanoparticles, antibodies, and protein-binding small-molecular-weight agents into tumors. Therefore, taking advantage of the SUPR effect after NIR-PIT may be a promising avenue to use a wide variety of nanodrugs in a highly effective manner.
