Effective combination therapy requires targeted co-delivery of multiple therapeutic agents via a well-defined and controllable assembly mechanism, which most reported strategies struggle to achieve. In this study, we designed a tumor acidity-driven transformable nanoparticle self-assembly using a drug-conjugated amphiphilic polymer (mPEG-PLA-Ce6), an acidity-sensitive polymer (PAEMA), and the CSF-1R inhibitor, sotuletinib (BLZ-945), by regulating the pKa and ratio of the acidity-sensitive material (denoted as Ce6SNP/B). The obtained tumor acidity-driven transformable Ce6SNP/B released BLZ-945 to deplete immunosuppressive M2-type tumor-associated macrophages predominantly localized in the perivascular regions of blood vessels. Simultaneously, tumor acidity-driven size shrinkage of Ce6SNP/B facilitated the deep penetration and tumor accumulation of photosensitizer Ce6 to enhance phototherapy, resulting in enhanced immunogenic cell death of tumor cells. Additionally, the acidity-sensitive material PAEMA has the potential to induce dendritic cell maturation. Thereby, the tumor acidity-driven transformable Ce6SNP/B achieved cancer photoimmunotherapy by targeting tumor cells and activating antigen-presenting cell-mediated anti-tumor immune effect.

Nucleic acid-based therapeutics hold great promise for cancer immunotherapy but remain limited by the lack of safe, stable, and effective delivery systems. Herein, we report a multifunctional siRNA vesicular delivery platform induced by sulfonium-containing random heteropolypeptoids. These polymers integrate bioinspired polypeptoids with permanent cationic sulfonium groups, enabling efficient nucleic acid complexation. Optimization of side chains yielded SG37OG36-O-Bn, which uniquely drives siRNA assembly into well-defined SG37OG36-O-Bn/siRNA vesicular nanostructures, termed PS siRNA, rather than conventional polymer–siRNA nanospheres. The resulting nanovesicles demonstrated remarkable storage stability, with high transfection efficiency preserved after long-term ambient storage. In tumor-bearing mice, PS siCD47 effectively silenced CD47, enhancing macrophage phagocytosis, dendritic cell maturation, and T cell-mediated anti-tumor immunity. Moreover, the vesicular architectures enabled co-encapsulation of catalase to decompose tumor-associated H2O2, remodel the immunosuppressive tumor microenvironment, achieved synergistic tumor inhibition with potent systemic anti-tumor immunity. This platform enables synergistic gene and enzyme therapy for next-generation cancer immunotherapy.

Messenger RNA (mRNA) vaccine is undoubtedly a medical breakthrough in drug development, however, its clinical application remains limited by inefficient delivery to target tissues and cells. In this study, we proposed a two-step screening strategy to optimize in vivo mRNA delivery system. First, we used the clinically approved cationic lipid, ionizable lipid and amphiphilic polymer to construct an initial library of lipid-polymer particles (LPP) with 60 various formulations for in vivo evaluation of their transfection efficiencies. Based on the results, we further constructed another library of 15 formulations to screen more effective LPPs. Then, the optimized LPP was selected and proved to be capable of effectively delivering mRNA to antigen-presenting cells (APCs), activating immune effector cells to trigger Th1/Th2 immune response, and promoting the formation of antigen-specific immune memory T cells. More importantly, LPP loaded with mRNA vaccine exhibited potent antitumor effects in both B16F10-OVA tumor model and human papillomavirus (HPV)-related TC-1 tumor model, exhibited comparable therapeutic activity to that of lipid nanoparticles (LNP) following intravenous injection. This study provides an innovative paradigm for the development of efficient mRNA delivery systems with high efficacy, safety, and clinical translation potential.

The therapeutic efficacy of chimeric antigen receptor (CAR)-T cell therapy in combating solid tumors remains constrained, primarily due to inadequate tumor infiltration and the immunosuppressive tumor microenvironment. Herein, we present a simple yet effective strategy for generating activated CAR-T-mimicking cells and enabling their magnetically guided migration into tumor tissues, thereby enabling a more potent and precise treatment of solid tumors. By functionalizing magnetic nanoparticles with anti-CD3 antibodies (aCD3) and anti-PDL1 antibodies (aPDL1), we have developed a magnetic bispecific nano-antibody (M-BiNanoAb), which effectively engages circulating T cells following intravenous administration and reprograms them into CAR-T-mimicking effector cells. Within this design, the aPDL1 and aCD3 moieties emulate the antigen-recognition domain and signaling domain of traditional CAR structures, respectively. Remarkably, the strategic application of an external magnetic field enables the precise navigation of these bioengineered T cells toward solid tumor regions, thereby facilitating the eradication of PDL1-overexpressing cancer cells. In preclinical models of solid tumors, this magnetically guided strategy for generating and manipulating CAR-T-mimicking cells demonstrated extraordinary antitumor activity, underscoring its transformative potential in advancing CAR-T-based therapies against solid malignancies.

Nanomaterials with intrinsic biological activity can directly participate in disease treatment, emerging as a pivotal focus in the development of next-generation therapeutics. In this study, we synthesized a library of cholesterol lipids bearing diverse tertiary amine head groups via a straightforward amidation reaction, then co-assembled them with amphiphilic polyethylene glycol-poly (lactic-co-glycolic acid) (PEG-b-PLGA) to formulate hybrid nanomaterials. Notably, the cholesterol derivative A3-Chol-formulated nanomaterials (A3-Chol@NP) polarized macrophages toward the pro-inflammatory M1 phenotype and enhanced phagocytosis of tumor cells. At the mechanistic level, A3-Chol@NP has been observed to preferentially interact with mitochondria in macrophages to produce mitochondrial reactive oxygen species (mtROS). This, in turn, activates ROS-NF-κB-iNOS and ROS-IRF5-IL-23 pathways, which have been identified as key factors in the macrophage polarization to M1-type. In the B16-F10 mouse melanoma model, A3-Chol@NP efficiently suppressed tumor growth via macrophage-mediated immunotherapy and completely blocked tumor progression when combined with anti-PD-L1 antibody.

Therapeutic cancer vaccines show great promise for de novo induction of antigen-specific T cell responses against tumors. However, weak coordination between innate immune activation and antigen delivery remains a major obstacle to vaccine efficacy. Here, we present a cholesterol azetidine derivative-assisted polymeric carrier, Aze-Chol NP, which stimulates innate immunity and primes tumor-specific CD8+ T cell responses. Mechanistically, Aze-Chol NP induces maturation and activation of dendritic cells (DCs) through the Toll-like receptor 9 (TLR9) pathway while simultaneously delivering protein or peptide antigens to DCs in lymph nodes. The Aze-Chol NP-based nanovaccines markedly inhibited tumor growth and prolonged survival in melanoma and human papillomavirus tumor models. Moreover, combining the nanovaccine with an anti-PD-L1 antibody produced a strong synergistic effect and long-term immune memory, achieving 80% survival beyond 100 days and complete rejection of tumor cell rechallenge. Overall, our study demonstrates that this TLR9-activating carrier provides an effective and straightforward strategy for developing potent cancer vaccines.

Endosomal escape is a limiting factor for the in vivo application of nucleic acid therapeutics and remains a major challenge in the development of drug delivery systems. In this study, we developed a pH-ultrasensitive membranolytic polymer (P(C6-BnX))-assisted delivery system (NPBnX) to facilitate siRNA endosomal escape and enhance the gene silencing efficiency. The incorporation of P(C6-BnX) imparts the siRNA delivery system with enhanced cellular uptake, effective membranolytic activity under endosomal pH conditions, and improved endosomal escape efficiency of siRNA. The significantly enhanced siRNA-mediated gene silencing efficacy was also confirmed in multiple tumor cell lines. Further investigations demonstrated that the delivery of siRNA targeting CD47 via this system effectively suppressed the expression of CD47 at both the cellular level and in vivo, thereby enhancing macrophage-mediated phagocytosis of tumor cells and promoting anti-tumor immune responses. This study provides a promising and viable strategy for the design and development of siRNA delivery systems.

Chimeric antigen receptor macrophages (CAR-Ms) are promising in solid tumor therapy due to their tumor-penetrating property and antigen-specific phagocytosis. However, current CAR-M therapy is limited by the low ex vivo proliferation of macrophages and the complexity of the engineering process. Generating CAR-Ms in vivo can overcome these challenges but still faces an M2-like pro-tumor phenotype polarized by immunosuppressive tumor microenvironment. Herein, we devise macrophage-preferential ionizable cationic lipid-assisted polymeric nanoparticles (iCLANs) to co-deliver mRNAs encoding interferon-gamma (IFN-gamma) and a CAR molecule, denoted as iCLANmCAR+mIFN-gamma, enabling in vivo engineering of CAR-Ms with a sustained M1-like phenotype. iCLANmCAR+mIFN-gamma can coexpress IFN-gamma and CAR in tumor-associated macrophages, thereby producing CAR-Ms capable of maintaining antitumor phenotype to effectively engulf tumor cells in an antigen-specific manner. Intravenous injection of iCLANmCAR+mIFN-gamma in EGFRvIII+ breast tumor and CD19+ B-cell lymphoma models directly generates EGFRvIII CAR-Ms or CD19 CAR-Ms within tumors, resulting in significant tumor growth inhibition and remodeling of the immunosuppressive tumor microenvironment. This study provides an efficient strategy for in vivo engineering of M1-like CAR-Ms for cancer therapy.

Selective eradication of tumor cells while sparing normal cells remains the ultimate goal of cancer therapy. Current molecularly targeted therapies exploit tumor-specific mutations or biomarkers to differentiate malignant from healthy cells. However, interpatient heterogeneity and the dynamic evolution of these targets limit their clinical applicability. To overcome this, we developed a targeted therapy strategy that exploits an intrinsic feature of melanoma: endogenous melanin. We integrated an HSP70 promoter-driven gasdermin E (GSDME) plasmid into a delivery system termed pCHG@iCLAN. Under 808 nm laser irradiation, endogenous melanin generates mild photothermal heating, which specifically activates the heat-inducible HSP70 promoter. This activation triggers GSDME and caspase-3-dependent pyroptosis selectively in melanoma cells, leaving normal cells and other cancer cell types unaffected. In B16-F10 murine models, this targeted therapy achieved 98% tumor inhibition and significantly prolonged survival. When combined with an immune agonist, it also elicited a durable immune memory that protected against tumor rechallenge. This work establishes a paradigm for using inherent tumor features, rather than variable molecular markers, to achieve precise targeted cancer therapy.

Cancer vaccines hold strong therapeutic promise, but their development is often hindered by the need to identify effective tumor antigens-a challenge amplified by tumor heterogeneity. Here, we report a tumor-confined nano-activator (iCLANTSP-SAg) that circumvents antigen dependence by driving localized expression of bacterial superantigens (SAgs) within tumors. This platform integrates tumor-specific promoter-driven SAg plasmids with lipid-assisted polymeric nanoparticles to enable efficient cytosolic delivery and tumor-restricted SAg production. The locally expressed SAgs elicit broad, antigen-independent activation of intratumoral CD4+ and CD8+ T cells through direct crosslinking of T-cell receptors and major histocompatibility complex class II molecules. By confining SAg expression to the tumor site, iCLANTSP-SAg minimizes systemic T-cell activation and off-target toxicity. When co-administered with anti-PD-1 antibodies, iCLANTSP-SAg eliminates established tumors in two-thirds of treated mice and induces durable immune memory that confers complete protection upon rechallenge. This strategy offers a potent and clinically translatable solution to the central challenge of antigen identification in cancer vaccine development.

Chimeric antigen receptor macrophages (CAR-Ms) are promising in solid tumor therapy due to their tumor-penetrating property and antigen-specific phagocytosis. However, current CAR-M therapy is limited by the low ex vivo proliferation of macrophages and the complexity of the engineering process. Generating CAR-Ms in vivo can overcome these challenges but still faces an M2-like pro-tumor phenotype polarized by immunosuppressive tumor microenvironment. Herein, we devise macrophage-preferential ionizable cationic lipid-assisted polymeric nanoparticles (iCLANs) to co-deliver mRNAs encoding interferon-γ (IFN-γ) and a CAR molecule, denoted as iCLANmCAR+mIFN-γ, enabling in vivo engineering of CAR-Ms with a sustained M1-like phenotype. iCLANmCAR+mIFN-γ can coexpress IFN-γ and CAR in tumor-associated macrophages, thereby producing CAR-Ms capable of maintaining antitumor phenotype to effectively engulf tumor cells in an antigen-specific manner. Intravenous injection of iCLANmCAR+mIFN-γ in EGFRvIII+ breast tumor and CD19+ B-cell lymphoma models directly generates EGFRvIII CAR-Ms or CD19 CAR-Ms within tumors, resulting in significant tumor growth inhibition and remodeling of the immunosuppressive tumor microenvironment. This study provides an efficient strategy for in vivo engineering of M1-like CAR-Ms for cancer therapy.

Interleukin-12 (IL-12) potently activates antitumor immune responses and compensates for the challenge of insufficient T cell activation and infiltration faced in immune checkpoint therapy. However, its clinical application is limited by severe systemic toxicity and the upregulation of PD-L1 on tumor cells during treatment. Here, we developed iLANpTyr-IL-12, a lipid-assisted nanoparticle system loaded with tyrosinase (Tyr) promoter-driven IL-12 plasmids, designed to achieve specific and sustained IL-12 expression exclusively in melanoma cells. The secreted IL-12 promoted dendritic cell maturation, T cell activation and proliferation, and IFN-γ secretion. Notably, iLANpTyr-IL-12 treatment upregulated PD-L1 expression on tumor cells, suggesting a negative feedback loop that supports the rationale for combination with anti-PD-L1 (αPD-L1). In a subcutaneous B16-F10 melanoma model, iLANpTyr-IL-12 monotherapy significantly enhanced tumor suppression and increased intratumoral infiltration of T cells and NKT cells, along with a decreased M2/M1 macrophage ratio. When combined with αPD-L1, it produced an enhanced antitumor effect, achieved a tumor growth inhibition rate of 83% and further augmented intratumoral CD8+ T cell infiltration. Additionally, no significant systemic toxicity or organ damage was observed. Collectively, these findings demonstrate that iLANpTyr-IL-12 enables tumor-specific IL-12 expression and, when combined with αPD-L1, provides a promising cytokine-immune checkpoint combination immunotherapy with reduced off-target toxicity.

Muscle genetic defects can lead to impaired movement, respiratory failure, and other severe symptoms. The development of curative therapies is challenging due to the need for the delivery of gene-editing tools into skeletal muscle cells throughout the body. Here, we use muscular fusogens (Myomaker and Myomerger) to engineer muscle-specific virus-like particles (MuVLPs) for the systemic delivery of gene-editing tools. We demonstrate that MuVLPs can be loaded with diverse payloads, including EGFP, Cre and Cas9/sgRNA ribonucleoproteins (Cas9 RNPs), and can be delivered into skeletal muscle cells via targeted membrane fusion. Systemic administration of MuVLPs carrying Cas9 RNPs enables skeletal muscle-specific gene editing, which excised the exon containing a premature terminator codon mutation in a mouse model for Duchenne muscular dystrophy (DMD). This treatment restores dystrophin expression in various skeletal muscle tissues, including the diaphragm, quadriceps, tibialis anterior, gastrocnemius, and triceps. As a result, the treated mice exhibit a significantly increased capacity for exercise and endurance. This study established a platform for precise gene editing in skeletal muscle tissues.

Local delivery systems bypass multiple delivery barriers and enhance the therapeutic agent concentration at the desired site via direct injection or post-operative implantation; however, their inability to refill therapeutic agents significantly limits their therapeutic efficacy. To address this issue, a refillable nanodrug-capturing system comprising a β-cyclodextrin (βCD)-modified alginate hydrogel (CdGel) and adamantane (Ad)-decorated nanoparticles (AdNPs) is developed herein. CdGel, locally administered at the disease site, captures and retains systemically injected drug-loaded AdNPs through host-guest interactions, enabling therapeutic agent refilling at the target site. Through the repetitive systemic administration of AdNPs, the refilling capacity of CdGel significantly enhances the anti-metastatic activity of the extracellular-targeted drug batimastat in both orthotopic and post-operative 4T1 breast tumor models. The proposed system facilitates the sequential administration of various drugs for combined cancer therapy and integrates the advantages of local and systemic delivery systems, enabling the enhanced drug accumulation and retention at the desired site.

The efficacy of checkpoint blockade immunotherapy for glioblastoma (GBM) is significantly influenced by the precise delivery of therapeutic agents that can penetrate the blood-brain barrier (BBB) and reprogram the tumor immune microenvironment. Conventional nanoscale carriers used for delivering immune checkpoint blockers are more likely to be internalized by tumor cells, leading to a loss of drug efficacy. This study presents a phosphatidylcholine (PC)-coated nanoparticle (PCNP) with an optimized PC ratio on its surface, achieving a balanced surface charge. This surface optimization minimizes nanoparticle-cell membrane interactions, reducing cellular uptake and thereby enhancing extracellular drug targeting efficacy. The constructed PC shell enabled PCNPs to penetrate the BBB mediated by choline transporters. The PC shell can attenuate interactions between PCNPs and cells, thereby preventing the internalization of PCNPs. Additionally, the poly-l-histidine core can undergo protonation in the acidic microenvironment, resulting in rapid disintegration of PCNPs and facilitating the quick release of the encapsulated CPI-444 (an extracellular adenosine receptor blocker) and temozolomide, inducing immunogenic cell death and blocking extracellular adenosine receptors to reverse the immunosuppressive feedback signaling pathway of the adenosinergic axis. This combination therapy has shown a novel therapeutic strategy for extracellular immune checkpoint blockade in GBM.

The mechanical properties of nanoparticles play a critical role in regulating their biological fate for drug delivery. This review analyzes the impact of nanoparticle elasticity on typical drug transport processes that have been explored in the past few decades, including blood circulation, tissue and tumor targeting, tumor penetration, cellular internalization, and drug release. Moreover, the relative mechanism is analyzed and discussed. This review provides useful clues for the design of new nanoparticles and better strategies for enhancing drug delivery and antitumor efficacy.

Immobilizing multiple types of monoclonal antibody (mAb) on nanoparticle surfaces is a promising approach for creating nanomedicines that emulate the functionality of multi-specific antibodies. However, the clinical translation of these multi-specific nano-antibodies (multi-NanoAbs) has been hindered by intricate fabrication procedures, inevitable attenuation in mAb affinity and insufficient carrier biosecurity. Here we develop a versatile nano-adaptor for immobilizing mAbs and construct multi-NanoAbs using a recombinant fusion protein that consists of Fc gamma receptor 1 and serum albumin, along with the biomedical polymer poly(l-lactide). Our findings demonstrate that fusion protein/polymer-based nano-adaptor is facilitated by FcγR1 on its surface to bind mAbs through receptor–ligand interactions rather than complex chemical conjugation and enables convenient and controlled construction of diverse multi-NanoAbs with efficacious therapeutic effects. We achieved large-scale production of humanized fusion protein/polymer-based nano-adaptor and confirmed the antitumour effectiveness of multi-NanoAb in humanized immune system mouse models, highlighting their prospects for clinical translation. A versatile nano-adaptor is used to construct multi-specific nano-antibodies for treatment of tumours in mice.

Acute lung injury (ALI) involves a vicious cycle of excessive reactive oxygen species (ROS) and inflammatory cytokines that current therapies have failed to address comprehensively. Here, we developed an inhalable biomimetic nanoplatform (M2M-SNPs) by coating ROS-responsive nanoparticles (SNPs) with M2 macrophage membranes (M2Ms), creating a dual-action platform that simultaneously scavenges ROS and neutralizes cytokines. More importantly, the M2M shell mediates targeted delivery to inflamed pulmonary tissues via natural homing properties, and the SNP core maintains receptor activity by preventing oxidative damage. In lipopolysaccharide (LPS)-induced ALI animal models, inhaled M2M-SNPs exhibited 2-fold-greater pulmonary accumulation than uncoated nanoparticles and achieved 58% ROS scavenging and 62-92% cytokine adsorption, significantly improving inflammatory infiltration and pulmonary edema. Transcriptome sequencing analysis further confirmed that M2M-SNPs mitigated ALI through systemic modulation of immune-inflammatory networks, particularly in IL-17 and TNF signaling pathways. This study provides a promising nanotherapeutic strategy for inflammatory respiratory diseases.