A dual-functional self-assembled nanoplatform integrating PDT and selective uPA inhibition for synergistic CRC treatment achieves short-term acute tumor ablation via PDT and sustained anti-metastatic potential via uPA inhibition within the tested observation windows, with favorable biosafety.
Abstract
Background: Colorectal cancer (CRC) is a major clinical challenge due to high metastasis and therapy resistance. Photodynamic therapy (PDT) offers precise tumor ablation but lacks sustained anti-metastatic activity. Peptidic urokinase-type plasminogen activator (uPA) inhibitors suppress metastasis but suffer from short half-life and poor tumor retention. This study aimed to develop a dual-functional self-assembled nanoplatform integrating PDT and selective uPA inhibition for synergistic CRC treatment. Methods: We designed and synthesized a conjugate by linking pyropheophorbide-a (PPA) with uPA-targeted cyclic peptide IG2, which self-assembled into nanoparticles (PINPs). Physicochemical properties, reactive oxygen species (ROS) generation, and uPA inhibitory activity were characterized. In vitro studies included cellular uptake, cytotoxicity, and invasion assays. In vivo therapeutic efficacy was evaluated in subcutaneous CT26 tumor models and lung metastasis models, with biosafety assessed by body weight monitoring. Results: PINPs exhibited uniform spherical nanostructure, prolonged blood circulation, and enhanced tumor accumulation via the enhanced permeability and retention (EPR) effect. Under 680 nm irradiation, PINPs generated robust ROS and induced tumor cell apoptosis. PINPs potently inhibited uPA activity and suppressed tumor cell invasion. In vivo, PINPs plus PDT achieved significant tumor growth inhibition (73.6%) and strong anti-metastatic efficacy (60.7%), superior to free IG2. No obvious systemic toxicity was observed. Conclusions: The dual-functional PINPs achieve short-term acute tumor ablation via PDT and sustained anti-metastatic potential via uPA inhibition within the tested observation windows, with favorable biosafety. This carrier-free self-assembly strategy provides proof-of-concept validation and a generalizable design paradigm for developing synergistic anti-metastatic nanotherapeutics against metastatic CRC.
Although strategic combination of cuproptosis and chemotherapy is emerging as a promising strategy against triple-negative breast cancer (TNBC), current drug delivery systems remain considerable challenges in achieving co-delivery of different formulas, such as complex nanocarrier design, limited drug loading capacity, and insufficient tumor targeting. Herein, cancer cell membrane-camouflaged, self-assembled nanoparticles (DCM@CCM) were fabricated for precision combination therapy against TNBC. In the strategy, the carrier-free self-assembled nanoparticles were one-pot fabricated by co-assembling copper ions (Cu2+), doxorubicin (DOX), and methotrexate (MTX) via hydrogen bonds, π-π stacking and metal-ligand coordination effect, followed by in situ camouflaging with cancer cell membranes. Benefiting from the homologous targeting effect, the developed DCM@CCM could specifically target tumor cells, promoting their cellular uptake. Following internalization into tumor cells, the DCM@CCM disassembled in response to a weakly acidic tumor microenvironment, releasing Cu2+, DOX, and MTX. Importantly, the Cu2+ was reduced to Cu+ by depleting intracellular glutathione, which not only activated cuproptosis but also catalyzed the endogenous hydrogen peroxide into highly toxic hydroxyl radicals via a Fenton-like reaction, resulting in mitochondrial dysfunction. Simultaneously, both DOX and MTX disrupted DNA synthesis to trigger cell apoptosis. Both in vitro and in vivo experiments indicated that DCM@CCM exhibited potent cytotoxicity against TNBC cells and effectively suppressed tumor growth in heterotopic tumor models with minimal side effects. Overall, our study not only provides a promising strategy for precision combination therapy against TNBC but also expands insight for developing nanoscale self-assembly-enabled nanomedicine.
Qian Liu, Xinyi Tao, Yawen Luo et al.· ACS Applied Bio Materials· 0 citations
Photodynamic therapy (PDT) holds promise for combination antitumor therapies by triggering immunogenic cell death (ICD). ICD is defined as the process by which tumor cells, upon death induced by external stimuli, convert from a non‑immunogenic to an immunogenic state, thereby mediating an anti‑tumor immune response in the host. But the poor aqueous solubility and inadequate tumor targeting of photosensitizers hinder their clinical translation. This study focuses on a novel BODIPY photosensitizer (M5) and aims to improve its antitumor efficacy via efficient tumor-targeted delivery and controllable release. Herein, we successfully synthesized iRGD-functionalized DSPE-PEG2000-iRGD and pH-sensitive HA-g-DEAP polymers, and further fabricated multifunctional M5/NMN/DEAP/iRGD-Lip liposomes via the thin-film dispersion method, which possess pH responsiveness and enhanced tumor-targeting ability. β-Nicotinamide Mononucleotide (NMN), a NAD + precursor, exerts a potent stimulatory effect on T-cell activation; 3-(Diethylamino)propylamine (DEAP) and hyaluronic acid (HA) can form pH-responsive HA-g-DEAP; the iRGD peptide (CRGDK/RGPDC), upon hydrolysis at its C‑terminus, exposes a motif that binds to neuropilin‑1 (NRP1), thereby conferring tumor‑targeting and tissue‑penetrating properties, endowing the liposomes with tumor-targeting and tissue-penetrating capabilities. M5 exhibits a high molar absorption coefficient of 5.33 × 104 M⁻¹ cm⁻¹ and a singlet oxygen quantum yield of 0.3854. In vitro cellular assays showed IC50 values of 104.1 nM and 72.68 nM in breast cancer MDA-MB-231 and 4T1 cells, respectively. Treatment with M5/NMN/DEAP/iRGD-Lip induced apoptosis rates of 56.84% and 55.6% in MDA-MB-231 and 4T1 cells, respectively. T-cell co-culture assays showed that M5/NMN/DEAP/iRGD-Lip increased the proportions of CD4+ and CD8+ T cells while reducing the proportion of regulatory T cells (Tregs) among CD4+ T cells. Collectively, the multifunctional M5/NMN/DEAP/iRGD-Lip liposomes integrate targeted delivery, pH-controlled release, and synergistic PDT-immunotherapy, effectively addressing key limitations of conventional photosensitizers. This work provides a promising nanoplatform for the development of novel combination therapies against breast cancer, laying a foundation for future preclinical and clinical translations.
Junwei Zhuang, Chen Guo, Xingming Ye et al.· Colloids and Surfaces B: Bio...· 0 citations
Rhodium-based nanophotosensitizers have recently emerged as promising platforms for near-infrared (NIR)-activated photodynamic therapy (PDT). However, their therapeutic potential remains largely restricted to reactive oxygen species-mediated cytotoxicity, limiting their ability to address the complex molecular drivers of tumor progression. Here, we report the therapeutic transformation of a previously validated transferrin-targeted rhodium nanophotosensitizer into a dual-action nano-therapeutic by incorporating the selective estrogen receptor modulator tamoxifen within the mesoporous silica matrix. The resulting system enables a combined therapeutic strategy that integrates endocrine therapy and light-triggered photodynamic action, providing a targeted approach for estrogen receptor-positive (ER+) breast cancer. Structural and functional characterization confirmed efficient drug loading, preservation of photodynamic activity, and maintained targeting capability. In vitro studies demonstrated enhanced cytotoxic efficacy compared with single-modality treatments, while intracellular quantification by LC-MS verified sustained tamoxifen delivery. To elucidate the molecular mechanisms underlying therapeutic efficacy, quantitative SILAC-based proteomics revealed coordinated disruption of oxidative stress defense, calcium homeostasis, mitochondrial energy metabolism, and tumor proliferation pathways. These findings indicate that the combined therapy induces a multifaceted cellular response involving both ROS-mediated damage and modulation of endocrine signaling. Together, this work establishes a mechanistically informed dual-modality nano-drug that expands the therapeutic scope of rhodium nanophotosensitizers beyond conventional PDT, providing a new strategy for targeted treatment of ER-positive breast cancer.
Andres Machuca, Alejandro Garcia Garcia, Estefanía García-Calvo et al.· Colloids and Surfaces B: Bio...· 0 citations
Therapeutic resistance in breast cancer, driven by tumor-intrinsic adaptive mechanisms and microenvironmental survival cues, remains a critical barrier to curative treatment. To address this dual challenge, we developed a redox-responsive polymeric micelle system (TPSP) functionalized with telmisartan for simultaneous targeting of angiotensin II type 1 receptor-overexpressing tumor cells and cancer-associated fibroblasts (CAFs). This platform co-encapsulates doxorubicin (DOX), a classic topoisomerase IIα (Topo IIα) poison, and aconitine linoleate (L29), a novel catalytic Topo IIα inhibitor with a distinct mechanism of action compared with conventional agents. The TPSP micelles exhibit dual therapeutic synergism: (1) L29 disrupts DNA replication through G1/S cell cycle arrest via Topo IIα catalytic inhibition, complementing DOX's DNA double-strand break induction to counter acquired resistance, and (2) telmisartan-mediated CAF depletion disrupts stromal-mediated drug resistance by eliminating metabolic symbiosis and biomechanical barriers. In vivo evaluations across resistant breast cancer models revealed superior tumor growth inhibition (>72%) with CAF ablation. This combinatorial nanomedicine strategy pioneers a paradigm shift in overcoming multidrug resistance by concurrently targeting tumor plasticity and microenvironmental protection, providing a clinically translatable blueprint for treatment-refractory malignancies.
Yunyu Xie, Xin Feng, Zhenyu Wang et al.· Biomaterials Science· 0 citations
Breast cancer remains a global health challenge with limited therapeutic options for metastatic cases. Cuproptosis, a copper-dependent cell death pathway, offers a novel anticancer strategy that is currently constrained by the poor solubility of copper complexes and lack of tumor selectivity. To overcome this limitation, a biomimetic nanoplatform was developed by co-encapsulating copper diethyldithiocarbamate (CuET) and a photothermal agent into epigallocatechin gallate-assisted nanoparticles, which was followed by coating with a hybrid cell membrane from cancer cells and programmed cell death protein 1 (PD-1)-overexpressing CTLL-2 cells. This strategy allowed synergistic induction of cuproptosis, mild photothermal therapy (PTT), and PD-1/programmed death-ligand 1 (PD-L1) checkpoint blockade. The nanoplatform exhibited excellent stability, photothermal efficiency, and tumor-targeting capability, resulting in responsive degradation within the tumor microenvironment. In vitro and in vivo studies using breast tumor models showed that these nanoparticles displayed potent cytotoxicity, causing immunogenic cell death and dendritic cell maturation. Transcriptomic analysis revealed significant enrichment of key biological pathways. Specifically, both immune-activation and cuproptosis-related pathways were markedly upregulated. The combination of mild PTT and PD-1/PD-L1 blockade enhanced CD8+ T-cell infiltration and established long-term immune memory. This multifunctional nanoplatform provides an integrated strategy for potentiating cuproptosis-augmented immunotherapy in breast cancer.
Glioma, a Grade-IV brain tumor, often exhibits functional suppression of P53 signaling due to aberrant stabilization of MDM2 by the deubiquitinase USP7, presenting a therapeutically exploitable vulnerability that remains under-utilised because of poor drug bioavailability and limited blood-brain barrier penetration. Here, we developed a rationally designed PLGA-based dual-loaded nanoformulation co-encapsulating USP7 inhibitor P5091 and P53-modulating polyphenol Resveratrol, to significantly attenuate the USP7-MDM2-P53 axis. Guided by synergy analysis, nanoparticles were formulated at an optimized molar ratio enabling controlled and sustained drug release with favourable physicochemical stability. Dual nanoencapsulation significantly enhanced synergistic cytotoxicity in glioma cells and 3D spheroids by inducing apoptosis through significant P53 restoration. Dual co-encapsulation improves pharmacokinetics and suppresses tumor growth with improved survival in orthotopic glioma model without any obvious vital organs histological damage. These findings highlight a mechanism-guided nanotherapeutic strategy for glioma treatment.
Sunny Kumar, Mrinal K. Ghosh· Nanomedicine: Nanotechnology...· 0 citations