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Despite a few clinical successes, the efficacy of cancer nanomedicines remains limited by rapid clearance by the mononuclear phagocytic system and poor permeation across the abnormal tumor vasculature. We previously showed that methyl palmitate nanoparticles (MPN) can safely and reversibly inhibit the phagocytic activity of immune cells for several hours, thereby improving tumor accumulation and the efficacy of systemically administered nanomedicines. Here, we demonstrate that, on a shorter time scale, MPN can induce vasodilation, introducing an additional mechanism to enhance the accumulation of therapeutic agents within malignant tissue. Upon internalization by macrophages and endothelial cells, MPN could potentially trigger the release of endogenous nitric oxide (NO), a key mediator of vasodilation, in a concentration-, and time-dependent manner. Following MPN administration, raster-scanning optoacoustic mesoscopy (RSOM) revealed vasodilation across multiple tissues, with a strong effect observed in tumors. To assess enhanced tumor accumulation, we injected 70 kDa fluorescent dextran and demonstrated via histology a markedly increased fluorescence signal exclusively in MPN-treated tumors compared to controls 24 h later. In addition, positron emission tomography (PET) imaging of 89Zr-labeled Feraheme nanoparticles showed significantly greater tumor accumulation after a 15 min MPN pretreatment. Finally, general serum biochemistry panels and histological analyses of major organs in healthy mice revealed that MPN did not induce observable short-term toxicity under the tested conditions (single or repeated MPN dosing). Overall, this study demonstrates that MPN-induced vasodilation occurring within minutes enhances intratumoral deposition of macromolecules and small nanoparticles. Together with their longer-term effects on phagocytosis inhibition, these findings indicate that MPN can improve therapeutic delivery through complementary, time-dependent mechanisms that increase tumor perfusion and vascular permeability.
Clinical translation of nanomedicines is greatly hindered by insufficient understanding of their in vivo process, yet a key challenge lies in quantifying the encapsulated versus free drug forms in tissues and cells. Herein, we present a facile, versatile anti-PEG single-chain variable-fragment antibody (PEG-scFv)-based method enabling quantitative measurement of both forms in various biofluids (e.g., interstitial fluid, cytoplasm). By this method, we map the in vivo process of PEGylated liposomal doxorubicin (sLip/Dox) at unprecedented resolution. In the bloodstream, doxorubicin remains largely encapsulated in liposomes (>99%). In liver as the main organ for drug elimination, less drug was distributed in the interstitium (>80% encapsulated) but more in liver cells (mainly in Kupffer cells) released in a time-dependent manner, accompanying doxorubicin transferred to hepatocytes most in free form by 12 h postinjection. After extravasation into tumors, there was a limited access of sLip/Dox to tumor cells, confining most of the drug in the interstitium mainly being encapsulated (more than 75%), and the internalized fraction underwent a gradual release process in both tumor-associated macrophages and tumor cells. These findings revealed that for sLip/Dox, which primarily underwent drug release intracellularly, cellular internalization rates could be the key factor in determining its in vivo performance. Given widespread PEGylation on developing nanomedicines and the cost-effectiveness of scFv production, PEG-scFv offers a broadly applicable tool for dissecting in vivo processes of nanomedicines to establish dose-effect relationships like small-molecule drugs, further to guide rational nanotherapeutic design.
The clinical translation of nanomedicine remains limited by the low and often unpredictable delivery efficiency of nanoparticles, largely arising from protein corona formation and its complex interplay with nanoparticle surface properties. In this context, strategies based on engineered protein coronas have emerged to improve control over nano-bio interactions. In this work, we investigate the interplay between an artificial protein corona formed using bovine serum albumin (BSA), the most abundant serum protein, and surface charge, one of the key determinants of interactions at the nano-bio interface, in shaping the biological behavior of silver nanoparticles (AgNPs). Negatively charged citrate-stabilized AgNPs and positively charged PAH-coated AgNPs were precoated with BSA and evaluated across three cell lines selected for their different sensitivities to nanoparticle surface charge. Our results show that BSA precoating markedly enhances nanoparticle stability in complex media, particularly for positively charged AgNPs, yet does not significantly alter the qualitative composition of the adsorbed protein corona or the cytotoxic responses elicited by the nanoparticles. Under serum-free conditions, however, BSA coating becomes essential for maintaining colloidal stability and enabling cellular interactions, thereby allowing AgNPs to exert their intrinsic biological activity. These findings demonstrate that although the artificial corona provides important colloidal stabilization, surface charge remains the dominant factor governing the biological behavior of AgNPs, further highlighting the need to elucidate the mechanisms underlying charge-dependent interactions at the nano-bio interface for the rational engineering of nanoparticle systems.
Nanoparticles show great potential for the management of acute kidney injuries (AKIs). It is well known that nanoparticle size plays a critical role in targeting renal tubules due to the pore structure and kidney filtration threshold (KFT) of the glomerulus. However, AKI can compromise the physiological barriers, which might render the established KFT (∼6 nm) and size laws under healthy conditions not applicable in AKI conditions. Reinvestigating the nanoparticle size effect on targeting injured kidneys is thus essential for developing efficacious AKI nanomedicine. Herein, using different-sized gold nanoparticles (AuNPs) ranging from 4 to 60 nm as a model of renal nanomedicine, we discovered that the KFT for AuNPs expands from ∼6 nm to ∼10-40 nm under rhabdomyolysis-induced AKI conditions. After loading curcumin, a natural antioxidant, into 5, 16, and 60 nm AuNPs to compare their efficacies against AKI, we found that 5 and 16 nm AuNPs both effectively delivered drugs into renal tubules, resulting in their equal therapeutic performance and significantly better efficacy than 60 nm ones, further validating the enlarged KFT under AKI. Our results provide valuable insights for size engineering of nanomedicine for better AKI management and enable a deeper understanding of AKI renal physiology at the nanoscale.
The pathogenesis of inflammatory bowel disease involves interconnected failures in epithelial integrity, immune cell migration, and inflammatory homeostasis in the system, making multifunctional therapeutic approaches a promising option. In this study, we developed a Bacillus licheniformis-templated nano-silver (Nano-Ag) composite within a tannic acid-iron (TA-Fe) metal-phenolic network and evaluated its therapeutic effects in TNBS-induced colitis. The composite exhibited favorable physicochemical properties, including a stable surface architecture and uniform nanosilver deposition, supporting its potential application as a biohybrid nanotherapeutic platform. In vivo treatment produced a dose-responsive protective effect, with the 1 mg formulation showing the most consistent response. This treatment regimen improved survival, reduced morbidity, alleviated diarrhea and fecal bleeding, lowered disease activity, preserved colon length, and significantly restored colonic histoarchitecture. Further histological examination revealed reduced inflammatory infiltration and improved hepatic morphology, indicating protective effects beyond the intestine. At the molecular level, therapeutic efficacy was associated with LEF-1 suppression, axin restoration, CXCR2 and CCR7 attenuation, and CCR5 recovery, consistent with coordinated normalization of Wnt-associated epithelial signaling and chemokine-guided immune trafficking. Serum profiling further revealed broad modulation of dysregulated chemokine networks and partial correction of biochemical disturbances associated with gut-liver and gut-kidney axis dysfunction. Collectively, these findings indicate that the engineered biohybrid nano-Ag platform does not act through a single anti-inflammatory mechanism, but rather through integrated remodeling of epithelial repair, inflammatory signaling, and systemic metabolic stress. Our study identifies TA-Fe-stabilized probiotic nano-Ag therapy as a promising multifunctional strategy for experimental colitis and provides a mechanistic framework for developing next-generation nanomedicines for inflammatory bowel disease.
Chromosome spreading is a key step in karyotyping and fluorescence-based analyses, yet it often suffers from limited dispersion, chromatid overlap, and morphological distortion due to operator-dependent variability. Here, we introduce an anisotropic, nanowrinkled polydimethylsiloxane (PDMS) substrate that enhances chromosome spreading by tuning droplet impact dynamics via anisotropic wetting and splashing-assisted transport. The uniaxial wrinkle geometry promotes lateral redistribution of the fixative droplet, leading to broader chromosome dispersion, stronger directional spreading, and improved chromatid separation compared with flat substrates. High-resolution atomic force microscopy reveals subtle but reproducible nanoscale surface modulations on chromosomes deposited on wrinkled PDMS, consistent with partial transcription of the underlying wrinkle relief during adsorption. The characteristic spacing of these modulations (∼300-500 nm) matches the wrinkle periodicity and is interpreted here as substrate-coupled nanoscale conformability, rather than as direct evidence of intrinsic chromatin organization. Collectively, these findings establish nanowrinkled PDMS as a practical, tunable platform for controlled chromosome deposition and further indicate that engineered nanoscale interfaces can elicit nanoscale conformability of metaphase chromosomes during adsorption.
RNA therapeutics are reshaping modern medicine, as exemplified by the rapid deployment of lipid nanoparticle (LNP)-based mRNA vaccines during the COVID-19 pandemic. However, the ability to reproducibly generate LNPs with precisely defined physicochemical properties at scale remains a critical challenge, particularly as the particle size and size distribution strongly influence biodistribution, cellular uptake, and therapeutic efficacy. Conventional mixing technologies ensure reproducibility but offer limited control over interfacial mixing and nanoparticle assembly, constraining both tunability and scalability. We present a hollow fiber membrane (HFM)-based platform that leverages dense arrays of nanoscale pores to mediate uniform, highly localized interfacial mixing, enabling controlled lipid self-assembly and RNA encapsulation. This nanopore-mediated architecture allows continuous, high-throughput synthesis of LNPs with tunable particle sizes, narrow size distributions, and high encapsulation efficiencies, with particle characteristics directly correlated to the membrane pore size. HFM-derived LNPs exhibit robust in vitro transfection and potent in vivo immune responses and are compatible with multiple lipid chemistries and nucleic acid payloads, including mRNA vaccine constructs. Together, this work establishes HFM-based nanopore assembly as a versatile and scalable approach for producing well-defined LNPs, with direct relevance to the current and future mRNA vaccine and therapeutic development.
Achieving dynamic control of the size of solid-state nanopores remains a significant challenge in single-molecule sensing. In this work, a flexible PDMS (polydimethylsiloxane) nanopore (3.9 μm thick) was fabricated by focused ion beam (FIB) milling and subjected to an applied transmembrane hydrostatic pressure in a KCl electrolyte, enabling real-time modulation of its aperture. The flexible PDMS nanopore exhibits nonmonotonic deformation in response to external pressure: initial constriction at low pressure, followed by re-expansion at higher pressure, with the smallest pore diameter achieved at an intermediate pressure. Leveraging this "constriction-dilation" gating behavior, we systematically regulated λ-DNA translocation─observing maximized ionic current blockade amplitude, prolonged dwell time, and optimal event frequency at the intermediate pressure regime. Furthermore, compared to a conventional rigid silicon nitride (SiN) nanopore, the flexible PDMS nanopore exhibited substantially reduced baseline current noise and enhanced signal-to-noise ratio (SNR), attributed to its low dielectric constant and mechanical stability. This work combines dynamic mechanical tuning and low-noise soft-material characteristics to realize a single-molecule detection platform with high sensitivity and selectivity for flexible, tunable nanopore sensing in DNA analysis and other biomolecular measurement applications.
Emerging nanomedicine strategies have established clinical translation and the ability to offset conventional therapeutic challenges. This study aimed to report the development and analysis of a nanoscale material that functioned as both a high-Z radiosensitizer and a prodrug delivery system. In this study, a nanoscale platform using two complementary radiosensitizing mechanisms was designed and developed to overcome nonspecific toxicity, which is a challenge in both chemotherapy and radiotherapy. High-Z physical radiosensitization was coupled with the inhibition of a critical DNA double-strand break repair mechanism, homologous recombination, through a lipid nanoparticle decorated with ultrasmall Hafnia nanoparticles and loaded with an amphiphilic prodrug. In vitro and in vivo studies demonstrated significant enhancement of therapeutic effects, cell killing, or tumor regression, respectively, versus radiation alone. These results support the use of this combination system as a practical strategy for maintaining treatment efficacy at modest doses of radiation and offset associated adverse effects.
Conventional chemotherapeutic agents are frequently limited by off-target toxicity and suboptimal therapeutic outcomes. Nanomedicines utilizing cell membrane camouflage provide a promising strategy for precise drug delivery and multimodal combination therapy. Herein, we designed a tumor-microenvironment-responsive cell-membrane-coated nanocomposite (designated as ZnO2@MO-D@Mn-CeO2@CM), which consists of a zinc peroxide (ZnO2) core encapsulated within a drug-loaded mesoporous organosilica (MO-D, where D is doxorubicin for the 4T1 breast cancer model and daunorubicin for the C1498 leukemia model) layer, with the mesopores gated by manganese-doped cerium oxide (Mn-CeO2) nanoparticles, and coated with a homologous cell membrane (CM). This nanocomposite enables the release of therapeutic components under acidic conditions and in the presence of elevated glutathione (GSH). It facilitates a combination therapy by integrating chemotherapy, enhanced chemodynamic therapy (CDT), ferroptosis induction, and immunomodulation. Our results demonstrate that this nanocomposite effectively suppresses the progression in 4T1 solid tumor and C1498 leukemia models, demonstrating its potential as a robust combinatorial strategy.
Metallic nanoparticles (NPs) enhance radiotherapy through photoelectric absorption and Auger electron cascades, yet the effective spatial range over which these low-energy electrons induce biological damage remains poorly defined. Quantifying nanoscale energy deposition is essential for rational therapeutic design and safe clinical translation. Here, we establish a self-assembled polyelectrolyte-nanoparticle-cell architecture enabling nanometer-precision control of NP-cell separation (25-100 nm) to directly probe distance-dependent radiation enhancement. Layer-by-layer assembly produced uniform interfaces confirmed by spectroscopy, ellipsometry, electron microscopy, atomic force microscopy, and microgravimetry. Using human microglial (HMC3) and diffuse intrinsic pontine glioma (SU-DIPG-IV) cells, we quantified intracellular reactive oxygen species generation and γH2AX-marked DNA double-strand breaks following 137Cs γ-irradiation. Cells positioned 25.9 nm from the NP layer exhibited significantly increased DNA damage relative to NP-free controls, whereas damage progressively decreased with increasing separation, yielding a 250% differential effect between 25.9 and 97.5 nm. Modality-dependent attenuation profiles were observed across γ-ray, X-ray, and electron irradiation. These findings define the effective nanoscale interaction radius governing NP-mediated Auger enhancement and establish a technique for the interrogation of light-matter interactions for therapeutic energy deposition.
The lack of safe, minimally invasive strategies for targeted drug delivery to the spinal cord remains a major barrier to treating neurodegenerative disease and spinal injury. Intrathecally administered macromolecules, including gene therapy vectors, rarely penetrate spinal tissue at therapeutically relevant concentrations due to cellular barriers at the cerebrospinal fluid-spinal cord interface. Here, we demonstrate the application of laser-activated perfluorocarbon nanodroplets (PFCnDs) for nonsurgical, spatially controlled delivery from the subarachnoid space into spinal cord parenchyma. Following intrathecal injection in rats, transdermal laser irradiation produced a 7-fold increase in photoacoustic signal within the spinal cord parenchyma compared to nonirradiated controls, confirming spatially selective intraparenchymal nanodroplet delivery. Codelivery of 500 kDa FITC-dextran demonstrated approximately 1 mm penetration into the dorsal horn at irradiated sites, while nonirradiated regions showed only superficial perivascular accumulation. Neither nanodroplets alone nor laser irradiation alone produced intraparenchymal delivery, indicating that vaporization-induced cavitation is necessary for penetration. The PFCnDs (∼300 nm diameter) were engineered with a lipid shell, perfluorohexane core, and near-infrared absorbing dye for transdermal activation at 1064 nm. These results demonstrate that laser-activated nanodroplets can deliver macromolecular cargo into the spinal cord via lumbar puncture and transdermal irradiation, without surgical exposure.
The emergence of biomembranes represents a pivotal leap in eukaryotic cell evolution, while the selective permeability of biological membranes necessitates active transport mechanisms, such as endocytosis, for macromolecular exchange to occur. Although signal transduction behaviors have been extensively investigated in both natural systems and artificial cellular models, the systematic coupling between membrane remodeling and transmembrane transport remains to be elucidated, posing a persistent challenge in developing a universal protocellular system for dynamic membrane architecture-mediated macromolecular transport. Herein, we have developed a nanoparticle-mediated, electrostatics-assisted strategy for transmembrane transport regulation in giant unilamellar vesicle (GUV)-based protocell systems from the perspective of micronano-scale synergistic interactions. This strategy utilizes functionalized negatively charged nanoparticles to synergistically integrate structure-induced contact forces and charge-mediated electrostatic repulsion, successfully inducing membrane invagination and vesicle formation, accompanied by nanoparticle transmembrane internalization, thereby triggering transport-like functionality in the protocells. Membrane property analysis and molecular dynamics simulations synergistically elucidated their interaction mechanisms, thereby confirming that the membrane remodeling process maintains structural integrity while promoting endocytosis-like macromolecular transport and signal transduction into protocells. Using erythrocytes as the structurally simplest natural cell model, we have further validated the universality and stability of this strategy in native phospholipid membrane systems. Overall, this nanoparticle-electrostatics synergy-based membrane regulation strategy provides a promising approach for membrane inward remodeling and macromolecular transmembrane transport studies in artificial cells.
Recent advances have demonstrated the application of microcavity-containing hydrogel microparticles, known as nanovials, for massively parallel and high-throughput screening of therapeutic T cell populations for adoptive cell therapies. Nanovial cavities coated with peptide-MHC (pMHC) or antigen tetramers selectively bind to their cognate T cell receptor (TCR) or chimeric antigen receptor (CAR) to activate T cells and capture secreted cytokines. However, binding of tetramers or recombinantly expressed antigen by T cells may not reflect physiological T cell activation or cytotoxicity, as the binding interface is not fully representative of the natural immunological synapse formed between T cells and professional antigen-presenting cells (APCs). Here, we leverage the recent discovery of an ESCRT- and ALIX-binding region (EABR) sequence to generate antigen-presenting vesicles and cancer-mimicking exosomes from standard HEK293T and Expi293F cell cultures. EABR-mediated vesicles present natural, full-length oncologically relevant membrane proteins embedded in lipid bilayers to functionalize the nanovial cavity with cell-like membranes. Hydrogel nanovials functionalized with the EABR-mediated vesicles show improved T cell capture of 1G4 T cells and enhanced induction of secretion in HER2 CAR-T cells compared to hydrogel surfaces functionalized with recombinantly expressed soluble proteins.
Burn wounds are highly susceptible to bacterial colonization, particularly by multidrug-resistant pathogens, which often result in refractory infections, excessive inflammation, and delayed healing. Herein, we report the rational design and construction of a multifunctional nano-antimicrobial agent, designated mPB@PHMB, through the co-assembly of mesoporous Prussian blue (mPB) with the polymer polyhexamethylene biguanide (PHMB). The resultant nanocomposite integrates potent antibacterial activity with the capacity to modulate the inflammatory microenvironment. Notably, mPB@PHMB exhibited pronounced bactericidal efficacy against methicillin-resistant Staphylococcus aureus (MRSA). Benefiting from the intrinsic multi-enzyme-mimetic activities of mPB, the nanocomposite efficiently scavenged intracellular reactive oxygen species, thereby alleviating oxidative stress-induced cellular injury. Transcriptomic profiling and protein analyses revealed that mPB@PHMB markedly suppressed the NF-κB pathway activation, attenuated lipopolysaccharide-induced M1 macrophage polarization, and reduced the production of pro-inflammatory cytokines. Biocompatibility assessments demonstrated negligible acute cytotoxicity toward skin fibroblasts and no overt toxicity in healthy C57BL/6 mice, underscoring the favorable safety profile of mPB@PHMB. Furthermore, in a murine model of MRSA-infected burn wounds, topical treatment with mPB@PHMB reduced the bacterial burden by approximately 90%. Concurrently, treatment with mPB@PHMB substantially decreased the proportion of M1-polarized macrophages, suppressed the expression of inflammatory cytokines, and alleviated the inflammatory milieu within the wound bed, thereby accelerating tissue repair and wound closure. Collectively, these findings establish mPB@PHMB as a promising nano-therapeutic platform that synergistically couples antimicrobial action with immunomodulatory functions, offering a robust and translatable strategy for the management of burn wounds complicated by multidrug-resistant bacterial infection.
Self-assembled peptide amphiphile (PA)-based nanostructures are promising biomaterials with their biocompatibility and highly tunable physical and chemical properties. Despite this promise, the interactions of PA nanostructures with biological systems have remained largely unexplored. Here, we investigate nano-bio interactions of self-assembled PA nanostructures using a series of PAs with varying amphiphilicities, achieved by tuning the hydrophilicity of the peptide backbone and payloads attached to them. We show that, while PAs with hydrophilic peptide backbones and payloads mostly disassembled in blood plasma, increasing overall hydrophobicity improved their stability. We also report that disassembled PAs can bind to blood biomolecules and binding shifted from albumin to lipoproteins with increasing hydrophobicity. In addition, we show that the membrane-binding affinity of PAs increases with increasing PA hydrophobicity, which also induces membrane disruption and cytotoxicity. The results of this study highlight the critical role of the overall amphiphilic balance of PAs on their interactions with biological molecules and cell membranes.
Innovations in therapeutic modalities and targeted drug delivery are two key approaches to improving the effectiveness of cancer treatment. With the development of therapeutic concepts and nanotechnology, recent research has shifted from tissue- or cell-level drug delivery to organelle-specific delivery to amplify therapeutic effects. Among these organelles, the endoplasmic reticulum (ER) has emerged as a particularly promising target because of its extensive membrane network, which serves as a major site for protein synthesis, lipid metabolism, calcium homeostasis, and intracellular signaling. The disruption of ER function can trigger severe cellular stress and apoptosis, providing a rationale for ER-targeted cancer therapy. However, traditional small-molecule drugs often exhibit poor ER specificity, rapid degradation, and limited intracellular accumulation, significantly restricting their therapeutic potential. In contrast, ER-targeting multifunctional nanoplatforms have demonstrated superior advantages, including enhanced stability, precise localization, controlled drug release, and multimodal therapeutic capabilities. In this Review, we systematically summarize the recent advances in ER-targeting nanotherapeutics and their applications in chemotherapy, phototherapy, immunotherapy, biotherapy, and combination therapy. Furthermore, we discuss the advantages, challenges, and mechanistic insights of these strategies, aiming to provide perspectives on accelerating the clinical translation of ER-targeting precision drugs.
Nasopharyngeal carcinoma (NPC) therapy faces challenges including severe off-target toxicity and immunosuppression. Here, a biomimetic nanogel vaccine is developed based on dual pH/reactive oxygen species (ROS)-responsive poly(N-vinylcaprolactam) (PVCL) nanogels (NGs) that were co-loaded with indocyanine green (ICG) and the endoplasmic reticulum stress-inducing drug toyocamycin, followed by coating with cancer cell membranes (CMs) or exosomes (Exos). Systematic comparison reveals that CM coating confers better homologous targeting, cellular uptake, and immunogenicity than the Exo coating. The optimized IT@PVCL-CM NGs enable tumor microenvironment-triggered drug release, combined chemotherapy and mild photothermal therapy, and synergistic induction of immunogenic cell death (ICD). In an NPC mouse model, the IT@PVCL-CM combined with laser irradiation significantly promotes dendritic cell maturation through the combination therapy-mediated ICD effect and the CM antigens, CD8+ T cell infiltration, and long-term memory T cell responses, effectively suppressing both primary and distant tumors via an abscopal effect. Additionally, ICG endows dual-modal fluorescence and thermal imaging for real-time tumor monitoring. This work presents the integration of dual-responsive PVCL nanogels, dual ICD induction strategies through combination of photothermo-chemotherapy, and biomimetic membrane cloaking for precision NPC theranostics and immune activation.
Atherosclerosis remains a major threat to human health due to unresolved plaque inflammation and macrophage dysfunction. Although stimuli-responsive nanocarriers (e.g., pH- or ROS-sensitive systems) have been exploited for targeted drug delivery, they suffer from modest signal gradients, spatial heterogeneity, or unpredictable release in atherosclerotic lesions. Herein, we report an esterase-responsive, carrier-free self-assembling prodrug nano-platform (LPNP) that exploits the unique pathological microenvironment of atherosclerotic plaques for targeted mTOR inhibition in lesional macrophages. The dual mTORC1/2 inhibitor Torkinib (PP242) was covalently conjugated to linoleic acid (LA) via a labile ester bond, enabling spontaneous nanoparticle formation without exogenous carriers. This ester bond remains stable during systemic circulation but is efficiently cleaved by intracellular esterases-highly active in macrophage-derived foam cells-enabling lesion-specific drug release. In vitro, LPNPs were readily internalized by macrophages, where they suppressed S6K phosphorylation, activated autophagy, reduced ROS levels, and upregulated ABCA1/ABCG1-mediated cholesterol efflux, thereby reducing lipid droplet accumulation and promoting M1-to-M2 repolarization. In an ApoE-/- mouse model, LPNP administration significantly reduced plaque burden, increased collagen deposition, and enhanced plaque stability without altering systemic lipid profiles or causing overt toxicity. Collectively, this esterase-responsive prodrug strategy couples carrier-free self-assembly with macrophage-associated intracellular activation, providing a promising approach to enhance lesional macrophage mTOR inhibition for atherosclerosis treatment.
Pulmonary delivery of lipid nanoparticle (LNP)-based mRNA vaccines offers a promising strategy for localized lung cancer immunotherapy, yet how distinct pulmonary administration routes determine cellular targeting and therapeutic efficacy remains poorly understood. Here, we systematically evaluate intranasal and intratracheal delivery of mRNA-LNP vaccines and reveal a route-dependent immunological mechanism governing lung-targeted tumor vaccination. Although intratracheal administration yields only a 2.6-fold increase in total pulmonary protein expression compared to intranasal delivery, it produces a striking 26.8-fold enhancement in functional mRNA transfection efficiency within alveolar macrophages, the dominant antigen-presenting cell population in the alveolar space. This selective targeting reprograms alveolar macrophages toward an activated antigen-presenting phenotype, promoting efficient antigen presentation, robust CD8+ T-cell responses, and superior prophylactic and therapeutic efficacy in pulmonary tumor models. Notably, local depletion of alveolar macrophages completely abolishes the antitumor protection conferred by intratracheal vaccination, establishing their indispensable role in mediating pulmonary mRNA vaccine efficacy. Together, these findings provide mechanistic insights into lung-targeted mRNA cancer vaccination driven by alveolar macrophage engagement, providing critical insights for the design of next-generation LNP-based nanomedicines for lung cancer immunotherapy.