Inspired by the non-transmembrane transfer of mitochondria in cell-to-cell communications, herein, we report an original exploration to accelerate mitochondrial intercellular transport, and its application to exogenous cargo delivery. We discover that deliberate PINK1-targeted mitophagy downregulation elevates mitochondrial transit capacity via multifaceted drivers-morphological adaptation, metabolic reprogramming, and respiratory enhancement. Capitalizing on this, we engineer high-speed mitochondrial vehicles for photosensitizer hitchhiking, with spatiotemporal tracking elucidating its dynamic intercellular transit and physiological impacts. Through mitochondria's communication network-tunneling nanotubes (TNTs), the mitochondria-photosensitizer cotransporter achieves reinforced intercellular delivery, thereby inducing deep tumor penetration and enhanced photodynamic killing. Our work establishes a transformative mitochondria-hitchhiking platform for overcoming biological barriers in drug delivery and provides mechanistic insights into manipulating intercellular organelle transport for therapeutic applications.

The efficacy of drug delivery, particularly for solid tumors, is severely hampered by a cascade of biological barriers-including dense extracellular matrix, high interstitial fluid pressure, and inefficient vascularization-that limit therapeutic penetration and distribution. Biological materials, such as cells, extracellular vesicles and organelles, serve as biocompatibility and targeted delivery vehicles, offering significant therapeutic potential. However, most current strategies emphasize multifunctional and biomimetic delivery systems designed to traverse the extracellular stroma, often overlooking intracellular transport pathways. Here, we demonstrate that mitochondria and their hitchhiked cargos are transported via tunneling nanotubes (TNTs), contiguous cytoplasmic bridges that interconnect cells. Oxidative stress plays a pivotal role in stimulating both TNTs formation and mitochondrial transfer. By leveraging TNTs as an intracellular highway, we achieved intercellular transport and deep tissue penetration of the photosensitizer IR780, which was hitchhiked onto mitochondria (designated as IR780/Mito). The intensity of near-infrared (NIR) light governs TNTs dynamics, either promoting formation or inducing cleavage, by modulating oxidative stress levels generated upon IR780 excitation. Under mild NIR irradiation, moderate oxidative stress enhances TNTs formation and facilitates IR780/Mito transfer, enabling efficient delivery directly into tumor cells. Conversely, intense NIR irradiation triggers excessive reactive oxygen species (ROS) production, leading to TNT disruption and subsequent blockade of IR780/Mito transport. These findings present emerging opportunities for exploration into the use of TNTs transshipment channel to realize controllable intracellular transport of different types of cargos, with broad and promising applications in the diagnosis and treatment of multiple diseases in the future. STATEMENT OF SIGNIFICANCE: This work presents a new strategy for delivering drugs deep into tumors by hijacking the body's own cellular "highways"-tunneling nanotubes (TNTs). Instead of designing synthetic nanoparticles, we load drugs directly onto mitochondria, the cell's energy units, which naturally travel between cells via TNTs. Using a safe near‑infrared light, we can precisely turn this delivery route on or off by controlling the level of cellular stress: mild light promotes transport, while strong light shuts it down. This approach leverages natural cell‑to‑cell communication to overcome physical barriers in solid tumors, offering a smart, biocompatible platform for improving precision cancer therapy and treating other diseases involving intercellular signaling.

Background and ObjectiveThe emerging N-acetylgalactosamine-small interfering RNA (GalNAc-siRNA) conjugates lead the way for liver-targeting delivery to exert gene-silencing therapeutic effects. To facilitate the drug development of GalNAc-siRNA, further detailed understanding of the key modality-specific mechanisms underlying the temporal discordance between pharmacokinetics and pharmacodynamics and how these processes can be extrapolated from animals to humans is needed.MethodsA mechanistic minimal physiologically based pharmacokinetic/pharmacodynamic (mPBPK-PD) model for an investigational new apolipoprotein C-III (APOC3)-silencing GalNAc-siRNA (RBD5044) was developed using available pharmacokinetic/pharmacodynamic (PK/PD) data. The aim was to explore hepatic-targeting delivery processes, the PK/PD relationship, and interspecies translation.ResultsFirst, multiple PK/PD datasets from mice were satisfactorily fitted using the mPBPK-PD model. Second, we translated the mice model to the monkey model, validated it, and then extrapolated from mice and monkeys to humans to simulate the PK/PD characteristics. We then mechanistically summarized and proposed the essential in vivo delivery processes of GalNAc-siRNA after subcutaneous administration (termed "ADUEB": Absorption [into system circulation], Disposition [distribution to liver target and elimination], Uptake [into hepatocytes], Escape [from endosome and lysosome compartments], and Binding [with argonaute2 to form RNA-induced silencing complex]). The targeting delivery coefficients of these processes achieved with the model using RBD5044 and the published data of another GalNAc-siRNA (fitusiran) quantitatively reflected the delivery efficiency and rate-limiting factors in targeted hepatocytes.ConclusionThis study successfully constructed the mPBPK-PD model and conducted interspecies extrapolation for a GalNAc-siRNA targeting APOC3. Promising quantitative insights into a hepatic-targeted GalNAc-siRNA delivery system are provided to characterize the unique temporal disconnection of PK/PD properties and evaluate the key in vivo delivery processes. It will promote model-informed strategies and quantitative mechanistic understanding to support efficient drug development, evaluation, and clinical application of this modality in the future.

Dying tumor cells regulated by immunogenic cell death (ICD) inducers are promising candidates for cancer vaccine development because of their comprehensive antigen spectrum. However, their limited immunogenicity and potential tumorigenicity hinder clinical translation. To address these challenges, a nano-orchestrator is developed that targets the endoplasmic reticulum (ER) stress, a critical pre-ICD event, to optimize the "precise dose" of ER stress. Using a clinical-range irradiation fluence (50‒200 J cm-2) with an 808 nm laser, the release of damage associated molecular patterns (DAMPs) and antigens are precisely regulated. A fluence of 150 J cm-2 (2 W cm-2 for 75 s) increases dendritic cell maturation and antitumor T cell proliferation, providing valuable clinical insights. The ER stress nano-orchestrator enhances both adjuvanticity and antigenicity via the protein kinase R-like endoplasmic reticulum kinase (PERK)-C/EBP homologous protein (CHOP) pathway to regulate ICD-induced DAMPs and promote tumor cell apoptosis. These optimized ER stress phototherapeutic dying tumor cells can serve as prophylactic vaccines, achieving a remarkable 100% success rate against tumor rechallenge in vivo. Additionally, the nano-orchestrator shows the potential to develop in situ therapeutic tumor vaccines when combined with anti-PD-L1 treatment, providing important insights into enhancing the efficacy of immune checkpoint regulators by modulating endogenous immune responses.

TGF-β is a crucial regulator in tumor microenvironment (TME), especially for myofibroblastic cancer-associated fibroblasts (myCAFs). The myCAFs can be motivated by TGF-β signaling to erect pro-tumor TME, meanwhile, myCAFs overexpress TGF-β to mediate the crosstalk between tumor and stromal cells. The blockade of TGF-β can break cancer-associated fibroblasts barrier, consequently opening the access for drugs into tumor. The TGF-β is a promising target in anti-tumor therapy. Herein, we introduced a two-stage combination therapy (TC-Therapy), including TGF-β receptor I inhibitor SB525334 (SB) and cytotoxicity agent docetaxel micelle (DTX-M). We found that SB and DTX-M synergistically inhibited myCAFs proliferation and elevated p53 protein expression in BxPC-3/3T3 mixed cells. Gene and protein tests demonstrated that SB cut off TGF-β signaling via receptor blockade and it did not arouse TGF-β legend compensated internal autocrine. On the contrary, two agents combined decreased TGF-β secretion and inhibited myCAFs viability marked by α-SMA and FAPα. TC-Therapy was applied in BxPc-3/3T3 mixed tumor-bearing mice model. After TC-Therapy, the α-SMA+/ FAPα+ myCAFs faded increasingly and collagenous fibers mainly secreted by myCAFs decreased dramatically as well. More than that, the myCAFs barrier breaking helped to normalize micro-vessels and paved way for micelle penetration. The TGF-β protein level of TC-Therapy in TME was much lower than that of simplex DTX-M, which might account for TME restoration. In conclusion, TGF-β inhibitor acted as the pioneer before nano chemotherapeutic agents. The TC-Therapy of TGF-β signaling inhibition and anti-tumor agent DTX-M is a promising regimen without arising metastasis risk to treat pancreatic cancer. The therapeutic regimen focused on TGF-β related myCAFs reminds clinicians to have a comprehensive understanding of pancreatic cancer.

Cancer progression and treatment-associated cellular stress impairs therapeutic outcome by inducing resistance. Endoplasmic reticulum (ER) stress is responsible for core events. Aberrant activation of stress sensors and their downstream components to disrupt homeostasis have emerged as vital regulators of tumor progression as well as response to cancer therapy. Here, an orchestrated nanophotoinducer (ERsNP) results in specific tumor ER-homing, induces hyperthermia and mounting oxidative stress associated reactive oxygen species (ROS), and provokes intense and lethal ER stress upon near-infrared laser irradiation. The strengthened "dying" of ER stress and ROS subsequently induce apoptosis for both primary and abscopal B16F10 and GL261 tumors, and promote damage-associated molecular patterns to evoke stress-dependent immunogenic cell death effects and release "self-antigens". Thus, there is a cascade to activate maturation of dendritic cells, reprogram myeloid-derived suppressor cells to manipulate immunosuppression, and recruit cytotoxic T lymphocytes and effective antitumor response. The long-term protection against tumor recurrence is realized through cascaded combinatorial preoperative and postoperative photoimmunotherapy including the chemokine (C-C motif) receptor 2 antagonist, ERsNP upon laser irradiation, and an immune checkpoint inhibitor. The results highlight great promise of the orchestrated nanophotoinducer to exert potent immunogenic cell stress and death by reinforcing ER stress and oxidative stress to boost cancer photoimmunotherapy.

Cancer stem cells (CSCs) characterized by self-renewal, invasiveness, tumorigenicity and resistance to treatment are regarded as the thorniest issues in refractory tumors. We develop a targeted and hierarchical controlled release nano-therapeutic platform (SEED-NPs) that self-identifies and responds to CSC and non-CSC micro-niches of tumors. In non-CSC micro-niche, reactive oxygen species (ROS) trigger the burst release of the chemotherapeutic drug and photosensitizer to kill tumor cells and reduce tumor volume by combining chemotherapy and photodynamic therapy (PDT). In CSC micro-niche, the preferentially released differentiation drug induces CSC differentiation and transforms CSCs into chemotherapy-sensitive cells. SEED-NPs exhibit an extraordinary capacity for downregulating the stemness of CD44+/CD24- SP (side population) cell population both in vitro and in vivo, and reveal a 4-fold increase of tumor-targeted accumulation. Also, PDT-generated ROS promote the formation of tunneling nanotubes and facilitate the divergent network transport of drugs in deep tumors. Moreover, ROS in turn promotes CSC differentiation and drug release. This positive-feedback-loop strategy enhances the elimination of refractory CSCs. As a result, SEED-NPs achieve excellent therapeutic effects in both 4T1 SP tumor-bearing mice and regular 4T1 tumor-bearing mice without obvious toxicities and eradicate half of mice tumors. SEED-NPs integrate differentiation, chemotherapy and PDT, which proved feasible and valuable, indicating that active targeting and hierarchical release are necessary to enhance antitumor efficacy. These findings provide promising prospects for overcoming barriers in the treatment of CSCs.

Immunotherapy has great promise in improving malignant tumor treatment. However, the efficacy of existing strategies is often limited by the immunosuppressive environment. Here, we demonstrate an in situ bionic immunoactivator, PLT-Bec1/DTA-1, with possessed natural advantages of platelets for tumor recruitment and activation, on which DTA-1 (CD357 monoclonal antibody) and Bec1 were tethered as combined immune boosters. PLT-Bec1/DTA-1, as a self-triggered release repository, can deliver the pre-tethered Bec1 and DTA-1 deeply through the secretion of platelet microparticles (PMPs), thereby cooperate tacitly and exhibit superiority in immune activation of dendritic cells (DCs) and T cells via autophagy inducibility, coupled with glucocorticoid-induced tumor necrosis factor receptor (GITR)-triggered T Reg suppression, remodeled the immunosuppressive network of tumor microenvironment. PLT-Bec1/DTA-1 promoted antigen presentation and T cell proliferation, and alleviated the low activity state of bone marrow-derived dendritic cells (BMDCs) in tumor suppressive environment. PLT-Bec1/DTA-1 inhibited tumor recurrence (5- and 13-fold lower of control group in tumor volume) and CD8 + T/T Reg ratio (6.3- and 8.8-fold vs. control group) in mouse tumor model after intravenous or subcutaneous administration. Also, PLT-Bec1/DTA-1 prevented tumor colonization in lung through in situ immune activation, and was slightly superior to the combined of Bec1 and PD-L1. Our findings highlight the promise of delivering immunostimulatory payloads via bionic carriers, eliciting automatic in situ activation of effector immune cells in tumor microenvironment for tumor eradication. All these results provide promising prospects into the application of immunoactivator in improving cancer synergistic immunotherapy to overcome the bottlenecks in clinic.

Polymer-protein systems have excellent characteristics, such as non-toxic, non-irritating, good water solubility and biocompatibility, which makes them very appealing as cancer therapeutics agents. Inspiringly, they can achieve sustained release and targeted delivery of drugs, greatly improving the effect of cancer therapy and reducing side effects. However, many challenges, such as reducing the toxicity of materials, protecting the activities of proteins and controlling the release of proteins, still need to be overcome. In this review, the design of hybrid polymer–protein systems, including the selection of polymers and the bonding forms of polymer–protein systems, is presented. Meanwhile, vital considerations, including reaction conditions and the release of proteins in the design process, are addressed. Then, hybrid polymer–protein systems developed in the past decades for cancer therapy, including targeted therapy, gene therapy, phototherapy, immunotherapy and vaccine therapy, are summarized. Furthermore, challenges for the hybrid polymer–protein systems in cancer therapy are exemplified, and the perspectives of the field are covered.

Despite the promising potential of cancer vaccine, their efficacy has been limited in clinical trials and improved methods are urgently needed. Here we designed a nanovaccine platform that contains dendritic cell derived exosomes carriers and patient-specific neoantigens for individualized immunotherapies. The nanovaccine exhibited convenient cargo loading and prolonged cargo transportation to the lymph nodes, followed by eliciting potent antigen specific broad-spectrum T-cell and B-cell-mediated immune responses with great biosafety and biocompatibility. Strikingly, delivery of neoantigen-exosome nanovaccine significantly prohibited tumor growth, prolonged survival, delayed tumor occurrences with long-term memory, eliminated the lung metastasis in the therapeutic, prophylactic and metastatic B16F10 melanoma as well as therapeutic MC-38 models, respectively. Additionally, exosome-based nanovaccine demonstrated synergistic antitumor response superior to liposomal formulation due to presence of exosomal proteins. Collectively, our research indicated improved strategies for cell free vaccines and suggested exosome-based nanoplatform for cancer immunotherapy and personalized nanotechnology. These findings represent a powerful pathway to generate individualized nanovaccine rapidly for clinical application.

Extracorporeal membrane oxygenation (ECMO), a kind of life support technology that can replace lung and heart function, is widely used in critical respiratory and circulatory exhaustion. Because of the serious diseases and the use of interventional catheters, patients receiving ECMO life support are often administrated with broad-spectrum antimicrobial agents, which increase the risk of fungal infection. Fungal infection during ECMO can increase mortality. How to effectively control fungal infection is a thorny problem faced by clinicians. During the treatment of ECMO, the patient's physiological status, ECMO oxygenation membrane, circulation pipeline and other factors may change the pharmacokinetic profiles of antifungal drugs, thereby affect the clinical efficacy of drugs. This artical reviews the pharmacokinetic characteristics of antifungal drugs during ECMO support, in order to provide references for clinical antifungal treatment.

作为一种可以替代肺和心脏功能的呼吸循环支持技术,体外膜氧合(extracorporeal membrane oxygenation,ECMO)近年来在各种危重症呼吸、循环衰竭中的应用逐渐增多[1-2].ECMO主要包括动力泵、氧合器、各种管路及监测系统等,静脉血通过体外膜肺氧合后再回输给静脉或动脉,实施肺保护性通气,从而使肺脏得到休息,肺功能得以恢复[3-4].ECMO的适应证是急性呼吸窘迫综合征(ARDS),部分新型冠状病毒感染病例发生严重ARDS,世界卫生组织(WHO)推荐常规治疗无效的危重型患者采用ECMO作为挽救性治疗[5].在新型冠状病毒肺炎大流行期间,ECMO在新型冠状病毒感染危重症患者救治中发挥了非常重要的作用.

This study reports the design of a novel excipient-free small molecule prodrug (DEX-ALA)-based nanoparticle (DEX-ALA/DTX) which combines (i) the ability to target dexamethasone (DEX) and docetaxel (DTX) simultaneously to an experimental solid tumor under the natural tumor-targeting properties with (ii) high drugs loading (49.3% DTX, 29.2% DEX, and 21.5% ALA), (iii) high stability (low premature release) in circulation, (iv) stimuli responsive disassembly and release DEX and DTX in tumor, and inducing immunogenic cell death (ICD) for immunotherapy. This concept is based on the inclusion of hydrophobic DTX into nanoparticles constructed by self-assembling of esterase-responsive prodrug of alpha-linolenic acid (ALA) dexamethasone conjugate. Computational and experimental evidences prove the recapitulation of nanoparticles structure and esterase-responsive release. In tumor compartment, DEX modulates tumor microenvironment, reduces interstitial fluid pressure (IFP) to promote DEX-ALA/DTX nanoparticles accumulating in tumor, and also suppresses inflammation induced tumor progression. DTX exerts its cytotoxicity effect to eliminate tumor cells and induces ICD to produce anti-tumor immune response. Both systemic administration and peritumoral administration of DEX-ALA/DTX nanoparticles display excellent tumor shrinkage on 4 T1 breast cancer models, but the histology investigation of the tumor biopsies evidences lung metastasis of peritumoral route. These findings give insight in overcoming tumor microenvironment obstacles to promote nanoparticles accumulation and provide a rational strategy to increase antitumor efficiency. This new excipient-free small molecule prodrug based nanotechnology platform is expected to have important applications in cancer therapy.

Tumor-associated macrophages (TAMs) are abundant in the tumor microenvironment and promote the tumor progression via multiple mechanisms. CD47 is overexpressed in most malignant tumors and acts as a "don't eat me" signal to inhibit phagocytosis. We utilized CRISPR/Cas9 technology to knock out CD47 to guarantee the long-lasting anti-tumor immune responses. However, the reprogrammed TAMs are vulnerable to the inhibitory cytokines and tend to be transformed back into TAMs, and CD47 blockade alone may not be sufficient to elicit effective immune responses. We combined CD47 blockade with an immune -activating cytokine IL-12 for synergistic anti-tumor efficacy by genetically engineering the tumor cells into factories of IL-12 to in situ reprogram TAMs. Firstly, we designed a selective responsiveness accelerated gene delivery system named HPT-PFs that was dual modified with hyaluronic acid (HA) and tumor microenvironment sensitive peptides (TMSP) to simultaneously deliver plasmids for CD47 knockout and IL-12 production. Due to tumor-specific transfection and excellent endosome escape ability of HPT-PFs vector, more than 27% of tumor cells lost CD47 expression after HPT-PFs mediated gene editing, thus eliciting the phagocytosis by macrophages. And higher than 500 ng/ml of IL-12 was produced by tumor cells after HPTPFs mediated pIL-12 expression, indicating the successful engineering of tumor cells into factories of IL-12. In melanoma-bearing mice models, drastic elevation of M1-polarized TAMs and secretion of inflammatory cytokines were observed when combined CD47 knockout with IL-12 production, which led to significant inhibition of tumor growth. Our study suggested that combination of CRISPR-mediated CD47 blockade with IL-12 production in tumor cells could synergistically promote macrophage-mediated immunotherapy, paving the way for CRISPR-based in situ engineering tumor cells for effective immunotherapy. (c) 2021 Published by Elsevier Ltd.

阻断免疫检查点程序性细胞死亡受体-1 (PD-1)或程序性死亡受体配体-1(PD-L1)可以增强效应T细胞的抗肿瘤活性.然而,许多患者对PD-1/PD-L1疗法缺乏反应.通过改善免疫抑制性肿瘤微环境(TME)以增强免疫检查点抑制剂的疗效已成为一种有前景的癌症治疗策略.本研究构建了具有基质金属蛋白酶(MMPs)响应能力的C-X-C趋化因子配体12 (CXCL12) siRNA与抗PD-L1肽的共给药脂质体(PD-L 1/siCXCL 12-Lp),联合siCXCL12的TME调控与抗PD-L1肽的免疫调节作用,以协同增强抗肿瘤免疫反应.动物实验方案经由北京大学生物医学伦理委员会审查通过.作者发现PD-L 1/siCXCL 12-Lp在体外(33.8％)和体内(15.5％)直接下调了CXCL12的表达,并有效提高了CD8+/Treg的比例(20.0％),这有利于抗PD-L1肽更好地发挥其免疫作用.联合治疗显著抑制了肿瘤生长(52.08％),并且具有良好的安全性,为癌症免疫治疗探索了新的思路.

Gliomas remain difficult to treat because of their metastatic and recurrent nature and the existence of the blood-brain barrier (BBB), which impedes drug delivery. Microglia, the resident macrophages in the CNS, can be recruited by gliomas and can penetrate the tumor. In this study, microglia (BV2 cells) are used as transport vectors to deliver paclitaxel for the treatment of glioma. To avoid paclitaxel toxicity in microglia, liposomes are first employed to isolate the drug from BV2 cells. Dipalmitoyl phosphatidylserine (DPPS), as an "eat me" signal, is doped into liposomes to amplify their phagocytosis by microglia. This study demonstrates that engineered microglia can cross the BBB, independently migrate toward gliomas, and transfer cargo to glioma cells. Of note, extracellular vesicles and tunneling nanotubes are found to offer unique modes of cargo transportation between microglia and glioma cells. In vivo, the engineered drug-loaded microglia has a high ability to target the brain, penetrate glioma, and suppress tumor progression, supporting the notion that the use of engineered microglia is a potential strategy for the treatment of glioma. These findings present new opportunities for exploration into the use of microglia as transport vectors to deliver therapeutic agents through specific membrane nanotubes and vesicles.

目的 对131I-c(RGD)2在原位荷脑胶质瘤动物模型中的靶向定位作用进行研究,以探讨其应用于脑胶质瘤诊断与治疗的可能性.方法 采用U87-MG人脑胶质瘤细胞接种在裸鼠右脑尾状核内,建立了原位荷脑胶质瘤动物模型;采用氯氨T法将131I标记c(RGD)2,于原位荷脑胶质瘤裸鼠中进行生物分布研究,计算131I-c(RGD)2在脑胶质瘤中的摄取率及肿瘤与正常脑组织摄取率的比值(T/NT).结果 给药后3 h及6 h,肾的摄取均为所有器官中最高;给药后3 h肿瘤未累及的脑组织的摄取率为(0.16±0.10)％ID/g,胶质瘤组织的摄取率为(0.41±0.26)％ID/g,脑胶质瘤中的摄取率显著高于正常脑组织摄取率(P值为0.025);给药后6 h肿瘤未累及的脑组织的摄取率为(0.08±0.04)％ID/g,胶质瘤组织的摄取率为(0.44±0.23)％ID/g,脑胶质瘤中的摄取率亦显著高于正常脑组织摄取率(P值为0.011);给药后3 h T/NT比值为3.36±1.86,给药后6 h T/NT比值增高至5.55±1.75.结论 c(RGD)2具有靶向颅内胶质瘤的能力,在胶质瘤的靶向诊断与治疗中具有潜力,但其在脑胶质瘤中的摄取率相对较低,有待于进一步提高.

Combination of several approaches which facilitates the blockade of multiple disease pathways has been proven highly effective in treatment of cancer cells and their microenvironment. Combinational gene therapy and chemotherapy via cationic liposomes holds great potential since it targets therapeutic agents synergistically increasing their selective accumulation at the tumor site and enhancing their efficacy allowing administration of lower doses of each agent, thus reducing their side effects. The present protocol describes the fabrication methods to obtain the reliable stealth liposomes coencapsulating vascular endothelial growth factor (VEGF) targeting small interfering RNA (siRNA), which can inhibit angiogenesis, and chemotherapeutic docetaxel (DTX), which can kill tumor cells efficiently. Besides, two receptor-specific peptides, specifically low-density lipoprotein receptor-related protein receptor (Angiopep-2) and neuropilin-1 receptor (tLyP-1), were attached on the stealth liposomes for tumor targeting and penetration.

Phototherapy and immunogenic cell death (ICD) are powerful strategies to fight cancer. However, their therapeutic outcomes are diminished by immunosuppressive and hypoxia microenvironment. Herein, a photo-based, immunomodulating and hypoxia-alleviated nanosystem, PDA-ICG@CAT-DTA-1, is proposed to achieve the synergism between phototherapy and immunotherapy. Catalase (CAT) and anti-GITR antibody (DTA-1) are loaded to photothermal agent and photosensitizer composed PDA-ICG nanoparticles. The PDA-ICG@CAT-DTA-1 exhibits intrinsic local hyperthermia and enhanced ROS generation in tumor, and abrogates tumor immune suppression. It results in reduction of intratumoral FOXP3(+) regulatory T cells (4.3-fold) and increase of CD4(+) effector T cells (1.5-fold) compare with the control, and promotes damage associated molecular patterns generation to reinvigorate ICD effect. The potent antitumor of PDA-ICG@CAT-DTA-1 is proved in 4T1 bilateral tumor-bearing mice, with inhibition ratio of 95.1% for primary cancers and 68.7% for abscopal cancers. Our findings highlight great promise of the constructed versatility nanosystem to fix bottlenecks for cancer therapy.

The blood-brain barrier (BBB) not only provides protection for the brain but also limits the diagnosis and treatment of neurological disease, because the therapeutic drugs must cross the BBB to reach the lesions. Intranasal (IN) delivery which provides both extracellular and intracellular pathways into the central nervous system (CNS) has been explored to deliver lots of drugs across the BBB to the brain and simultaneously decrease the hepatic first-pass metabolism and gastrointestinal adverse effects. Rivastigmine, a long-acting reversible and noncompetitive acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitor, has been used for treatment of patients with mild to moderately severe Alzheimer’s disease (AD). In order to increase rivastigmine distribution in the brain and proceed to enhance pharmacodynamics via intranasal delivery, the protocol to formulate rivastigmine liposomes (Lp) and cell-penetrating peptide (CPP) modified liposomes (CPP-Lp) are introduced, including the preparation and evaluation. The permeability of two rivastigmine liposomes across the BBB by murine brain microvascular endothelial cells model in vitro is described. The drug distribution and pharmacodynamics effect in vivo exhibit that rivastigmine liposomes could significantly improve retention and inhibit activities of AChE and BuChE in CNS regions especially in cortex and hippocampus. It was concluded that rivastigmine liposomes especially CPP-modified liposomes have great potential to improve the brain delivery and enhance pharmacodynamics with respect to the BBB penetration and nasal mucosa to brain route.