Engineering antigen as photosensitiser nanocarrier to facilitate ROS triggered immune cascade for photodynamic immunotherapy
Graphical abstract
Antigen is spatially packaged as photosensitiser nanocarrier to construct an immunogenic and photocytoxic nanoplatform (MON) that is able to amplify the immune response during the PDT. By successfully eliciting abundances of CD8+ T cells, MON completely ablates the B16-OVA tumor and provokes a long-term antitumor immune memory effect, which powers the photodynamic therapy into an immunotherapy.
Introduction
Photodynamic therapy (PDT) is a clinically approved approach that can exert the selective cytotoxic activity toward malignant tumor cells upon exogenous light [1,2]. It encompasses the advantages of minimally systemic toxicity for localized light irradiation and low resistance occurrence with repeated treatments [3,4]. In the implement of PDT, photosensitizers can be excited by visible light in the presence of oxygen to form reactive oxygen species (ROS), which possess cytotoxic properties for inducing cell apoptosis or necrosis, microvasculature shutdown and immune response [5,6]. During the past 30 years, PDT has been exploited increasingly in the treatment of a variety of solid tumors, including brain, skin, breast, cervix and ovarian etc. [[7], [8], [9], [10]].
Recently, it has been well documented that the immune response usually accompanied within the PDT process [11,12]. Among those PDT concurrent immune responses, ICD is described as a death process that can trigger immune response and complement the PDT effect [[13], [14], [15], [16], [17]]. Evidence has also been accumulated that PDT generated ROS can alter the tumor microenvironment and recruit inflammatory and immune mediators, favourable to prime immune response [11,18]. During the priming process, tumor-associated antigens originating from tumor debris are released and captured by antigen-presenting cells (APCs) to consequently activate tumor specific CD8+ T cells [[19], [20], [21]]. Damage-associated molecular patterns (DAMPs) released during PDT process act as immunoadjuvant to stimulate the APCs maturation [22,23]. Another benefit is the recent discovery of ROS capable of improving antigen cross-presentation (AC) efficiency necessary for CD8+ T cells generation [24]. Mechanistic study showed ROS could mediate alkalization of phagosomal pH due to the consumption of protons intracellularly, which prevents antigen degradation by the acidic lysosomal proteases [25,26]. Such a behavior facilitates the processing of antigen by major histocompatibility complex class I (MHC-I) in APCs, and subsequently enhances CD8+ T cell responses for cancer immunotherapy [27,28].
Despite of the abovementioned encouraging immune responses, PDT is far from being perceived as an effective immunotherapy for eliminating tumor or metastatic tumor for the limited effectiveness of immune response [29]. This is partially evidenced by the fact that PDT induced tumor lysates produce little immunotherapy effect [30]. To potentiate immunotherapy effect, PDT has to resort to integrate other complementary therapies to achieve synergistic therapy. Incorporation of immune checkpoint inhibitor is frequently employed, for example the inhibitor of indoleamine 2, 3-dioxygenase (IDO)[14], PD-1/PD-L1 [31] and CTLA-4 [32]. However, a prerequisite for checkpoint immunotherapy relies on the pre-establishment of adaptive immune response, i.e., the existence of CD8+ T cell. Additionally, the high cost and side effect of the combination also complicate the application in clinic [33].
To amplify the antitumor immune response during the PDT, this study focuses on spatially packaging antigen ovalbumin (OVA) as photosensitiser Ce6 nanocarrier to construct an immunogenic and photocytoxic nanoparticle. The Ce6 equipped antigen nanoplatform is expectedly to synergistically integrate the benefit of ROS mediated ICD and AC effects[25,26], and consequently augment the immune response. Onto the antigen nanocarrier, the particle surface will be further camouflaged with the cancer cell membrane to facilitate targeting efficiency towards tumor site by homophilic targeting [[34], [35], [36], [37]]. In this fashion, the camouflaging nanoparticles should be able to selectively accumulate in tumor site and exert a photocytoxic effect to produce tumor-associated antigens under laser irradiation. The produced tumor-associated antigens as well as MON were captured by APCs in situ then migrating to tumor draining lymph nodes (dLNs). The spatial antigen nanopackage and the colocalization with photosensitiser by structural organization are expected to cooperatively improve antigen cross-presentation. B16-OVA tumor bearing mice (expressing ovalbumin) are then employed to evaluate the efficacy of antitumor immune cascade leading to photodynamic immunotherapy. The role of the amplified immune effect will be discriminated from photodynamic effect by using non-OVA expressing tumor bearing mice as the control.
Section snippets
Materials
The photosensitiser (PS) Chlorin e6 was obtained from J&K Scientific, Ltd. Fetal bovine serum (FBS), RPMI Media 1640 (RPMI 1640), penicillin-streptomycin and trypsin were supplied by Gibco Invitrogen. 4, 6-diamidino-2-phenylindole (DAPI), WST-1, prestained color protein ladder, 1 × EDTA-free protease inhibitor and 2′, 7′-dichlorofluorescein diacetate (DCFH-DA) were obtained from Beyotime Institute of Biotechnology. Paraformaldehyde (4%) was obtained from DingGuo Chang Sheng Biotech. Ovalbumin
OVA antigen nanoparticle synthesis, Ce6 loading, cancer cell membrane camouflaging and structural characterization
The widely employed model antigen, OVA[39], was chosen in this study. We attempted to assemble antigens into nanoparticle by avoiding using any other conventional nanocarrier, to purposefully maximize the content of antigen. One OVA molecule contains four free thiols which lays the basis for spatially packaging OVA antigens themselves into nanoparticles with disulfide bond network between OVA molecules. However, a pretreatment of OVA solution by SDS is imperative since most of those free thiol
Conclusions
In this study, we have developed an integrative approach to achieve photodynamic immunotherapy by ROS triggered antitumor immune cascade. In this approach, OVA was nano-packaged by establishing an intermolecular disulfide bond network between antigen, which was employed as photosensitiser nanocarrier and subsequently coated with cancer cell membrane. The membrane coated OVA nanoparticle loading Ce6 (MON) exhibited the advantages of homophilic targeting, prolonged circulation time, promoting
Author statement
Huaiji Wang and Yongyong Li conceived the project and designed the experiments. Huaiji Wang performed most of the experiments and statistical analyses of the data. Kun Wang, Lianghua He and Ying Liu helped with animal experiment. Kun Wang assisted with data interpretation of immune assays. Huaiji Wang wrote the original manuscript, with edits by Yongyong Li and Haiqing Dong, and reviewed by all authors. Yongyong Li and Haiqing Dong supervised the whole project.
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgements
This work was financially supported by research grants from the National Natural Science Foundation of China (NSFC51773154, 31771090, 51473124 and 81571801), Shanghai Natural Science Foundation (17ZR1432100), the Fundamental Research Funds for the Central Universities (22120180062) and Young Hundred-Talent Program of Tongji University.
References (48)
- et al.
Photodynamic nanomedicine in the treatment of solid tumors: perspectives and challenges
J. Contr. Release
(2013) - et al.
Cytokines in immunogenic cell death: applications for cancer immunotherapy
Cytokine
(2017) - et al.
Oxygen-boosted immunogenic photodynamic therapy with gold nanocages@manganese dioxide to inhibit tumor growth and metastases
Biomaterials
(2018) - et al.
Self-adjuvanted nanovaccine for cancer immunotherapy: role of lysosomal rupture-induced ROS in MHC class I antigen presentation
Biomaterials
(2016) - et al.
NIR-responsive cancer cytomembrane-cloaked carrier-free nanosystems for highly efficient and self-targeted tumor drug delivery
Biomaterials
(2018) - et al.
A biomimetic cascade nanoreactor for tumor targeted starvation therapy-amplified chemotherapy
Biomaterials
(2019) - et al.
Fluorine-18 labeled mouse bone marrow-derived dendritic cells can be detected in vivo by high resolution projection imaging
J. Immunol. Methods
(2002) - et al.
A new sol–gel silica nanovehicle preparation for photodynamic therapy in vitro
Int. J. Pharm.
(2010) - et al.
Nanoparticles in photodynamic therapy
Chem. Rev.
(2015) - et al.
Overcoming the Achilles’ heel of photodynamic therapy
Chem. Soc. Rev.
(2016)
Multitriggered tumor-responsive drug delivery vehicles based on protein and polypeptide coassembly for enhanced photodynamic tumor ablation
Small
Photodynamic therapy of cancer. Basic principles and applications
Clin. Transl. Oncol.
Mitochondria-targeting magnetic composite nanoparticles for enhanced phototherapy of cancer
Small
Supramolecular photosensitizers rejuvenate photodynamic therapy
Chem. Soc. Rev.
Nanocomposite-based photodynamic therapy strategies for deep tumor treatment
Small
Recent progress in near infrared light triggered photodynamic therapy
Small
Photodynamic therapy for cancer
Nat. Rev. Canc.
Photodynamic therapy of cancer: an update
Ca-Cancer J. Clin.
Near-infrared-triggered photodynamic therapy with multitasking upconversion nanoparticles in combination with checkpoint blockade for immunotherapy of colorectal cancer
ACS Nano
Immunogenic cell death and DAMPs in cancer therapy
Nat. Rev. Canc.
Enhanced immunotherapy based on photodynamic therapy for both primary and lung metastasis tumor eradication
ACS Nano
Nanocarrier-mediated chemo-immunotherapy arrested cancer progression and induced tumor dormancy in desmoplastic melanoma
ACS Nano
Stimulation of anti-tumor immunity by photodynamic therapy
Expet Rev. Clin. Immunol.
Photodynamic therapy and anti-tumor immunity
Laser Surg. Med.
Cited by (66)
Adoptive cell therapy for solid tumors beyond CAR-T: Current challenges and emerging therapeutic advances
2024, Journal of Controlled ReleaseNanoplatform-enhanced photodynamic therapy for the induction of immunogenic cell death
2024, Journal of Controlled ReleaseMultifunctional tadpole-like bimetallic nanoparticles realizes synergistic sterilization with chemical kinetics and photothermal therapy
2023, Applied Catalysis B: EnvironmentalCitation Excerpt :For example, the antibacterial nanomotors based on lysozyme developed by Kiristi et al. [29] can promote the interaction between enzymes and bacteria and prevent the aggregation of dead bacteria, thereby greatly enhancing the bactericidal ability. Peng et al. [30] prepared NO-driven nanomotors for bacterial biofilm elimination and endotoxin removal for the treatment of infected burn wounds with comprehensive anti-biofilm and anti-inflammatory effects. Inspired by this, we developed multi-purpose nanozyme TNPs enabling simultaneous PTT, peroxidase-like catalytic activity, glutathione peroxidase-like activity, and catalase-like activity.