Nanoparticle-mediated inhibition of Yes-associated protein prevents corneal scarring after traumatic injury

Albumin-hitchhiking self-assembly full-API nanoparticles for imaging-guided photodynamic potentiating tumor immunotherapy.

ABSTRACT Hepatitis resulting from alcoholic liver disease (ALD) is an important risk factor for liver cancer. It is essential to understand the role of ALD to prevent liver cancer. Zinc finger E‐box binding homeobox 1 (ZEB1) is a well‐known transcription factor that can trigger tumorigenesis. However, how ZEB1 affects liver hepatitis related to ALD remains unclear. We used western blot, immunohistochemistry, and quantitative real‐time polymerase chain reaction to detect ZEB1, IL‐1β, MCP‐1, and YAP expression in the liver tissue of a mouse ALD model and in L02 cells treated with alcohol. We used flow cytometric analysis to detect the apoptosis ratio of L02 cells. Levels of IL‐1β and MCP‐1 increased in the liver tissue of the mouse ALD model, which indicated that ALD triggered hepatic inflammation. ZEB1 was upregulated in the liver tissue of the mouse ALD model and in the L02 cells treated with alcohol. We also examined how ZEB1 regulates liver hepatitis. The results showed that levels of IL‐1β and MCP‐1 were increased by ZEB1. Knockdown of ZEB1 decreased the ratio of apoptotic cells. Moreover, the pattern of YAP expression was completely consistent with the ZEB1 expression pattern, and we verified that ZEB1 unregulated the YAP1 level. Finally, we used verteporfin (VP), an inhibitor of YAP, to treat ZEB1‐overpressing L02 cells. We found that VP suppressed the level of inflammation in alcohol‐treated L02 cells induced by ZEB1 overexpression. ZEB1‐YAP‐IL‐1β/MCP‐1 may be a critical pathway in ALD development. ZEB1 is a crucial mediator of alcoholic liver disease.

Background: The combination of photodynamic therapy (PDT) and immunotherapy has been explored as an innovative approach to enhance efficacy against tumors. However, PDT shows limited effectiveness in treating deep-seated tumors, as light and lasers do not sufficiently penetrate tissue. Methods: Herein, we introduced a nanocarrier (NPVR) via self-assembly, using an amphiphilic copolymer to co-deliver the hydrophobic photosensitizer verteporfin (VP) and the immunoadjuvant imiquimod (R837). Results: Our X-ray-induced photodynamic therapy (X-PDT) mechanism induced NPVR to generate a large amount of cytotoxic reactive oxygen species (ROS), which directly killed cancer cells. Moreover, the released R837 facilitated immunogenic cell death following the X-PDT process and promoted the maturation of dendritic cells (DCs), thereby eliciting immune responses against malignant triple-negative breast cancer (TNBC). In animal experiments, the combined therapy using NPVR showed a tumor growth inhibition rate of ~70%. Conclusions: This novel strategy opens new avenues to designing next-generation nanomedicines for use in immunotherapy and other combination therapies.

ABSTRACTWet age‐related macular degeneration (wAMD), characterized by pathological choroidal neovascularization (CNV), is a leading cause of irreversible vision loss in the elderly. The current standard treatment—anti‐vascular endothelial growth factor (VEGF) therapy—effectively manages neovascularization in many patients. However, some experience suboptimal responses, and frequent intravitreal injections raise safety concerns. Photodynamic therapy is another effective option for treating wAMD, but it can lead to an increase in reactive VEGF after the procedure, resulting in CNV recurrence. In response to these challenges, we propose an integrated approach that combines a DNA nanoflower VEGF degrader with photodynamic therapy. The DNA nanoflower consists of numerous aptamer‐based lysosome‐targeted chimaera (LYTAC) units, which drive extracellular VEGF in the lesion area to the lysosome for degradation. Simultaneously, the DNA nanoflower acts as a carrier for verteporfin (VER), a clinically used photosensitizer. The resulting nanoflower, named NF@VER, generates reactive oxygen species under near‐infrared light to induce endothelial cell death. These combined effects on endothelial cells effectively block VEGF‐induced CNV in vivo, without causing noticeable side effects. Overall, this innovative approach presents a precise and effective strategy for treating wAMD, reducing the risk of VEGF reactivation‐induced CNV recurrence, and minimizing the systemic side effects associated with photodynamic therapy.

Background: Hepatocellular carcinoma (HCC), the major form of primary liver cancer, contributes markedly to cancer-related mortality worldwide and remains a serious global health concern, particularly affecting individuals with underlying chronic liver disorders. In hepatocellular carcinoma, insufficient radiofrequency ablation (iRFA) has been reported to drive local tumor relapse and distant spread, possibly by aggravating the immunosuppressive features of the tumor microenvironment. The present work seeks to clarify the underlying pathways driving the development of an immunosuppressive milieu after RFA and to identify potential therapeutic approaches to counteract this process. Methods: An injectable hydrogel composed of quaternized chitosan (QCS) and tannic acid (TA) was constructed to encapsulate verteporfin (VP), a well-established photosensitizer that has been clinically applied for treating neovascular retinal disorders such as age-related macular disease. Beyond its ophthalmologic application, VP has recently been reported to display anti-tumor activity through inhibition of oncogenic regulators such as Yes-associated protein (YAP), indicating its potential utility in cancer therapy. This hydrogel formulation is designed to target residual tumor tissue post-RFA, providing localized delivery and sustained release of VP to enhance anti-tumor immune responses. Results: Our findings identified YAP activation as a critical mediator of immunosuppression in residual tumors following RFA. Pharmacological inhibition of YAP significantly reduced the infiltration of myeloid-derived suppressor cells (MDSCs) and effectively reversed the immunosuppressive microenvironment conditions. Furthermore, the QCS/TA hydrogel enabled sustained local release of VP, resulting in enhanced antitumor immune responses via MDSC suppression. When administered as an adjuvant therapy following suboptimal RFA, the hydrogel markedly inhibited the progression of residual tumors, highlighting its therapeutic potential in improving post-RFA outcomes. Conclusion: Collectively, our data suggest YAP pathway inhibition as a promising immunomodulatory strategy to complement RFA in HCC management. This work demonstrates that the QCS/TA hydrogel-based delivery system can remodel the tumor immune milieu to overcome immunosuppression and delay post-ablation tumor recurrence, supporting its potential as a translational drug delivery strategy.

The Hippo signaling pathway plays a central role in regulating cell growth, and dysregulation of its downstream effector Yes-associated protein (YAP) leads to tumorigenesis. Verteporfin (VP), a clinically approved drug, inhibits YAP-TEA domain (TEAD) complex formation, yet its binding mechanism remains unclear. In this study, we conducted a comprehensive in silico analysis of all 4 VP isomers within the context of the full-length YAP-TEAD complex. The complex structure was modeled using AlphaFold2 multimer, which provided sufficient accuracy for docking simulations despite incomplete experimental data on YAP. Docking calculations were performed against 2 grids, one centered on a predicted druggable pocket and the other on the YAP-TEAD interface. A total of 304 poses were generated, and the top-scoring 100 were clustered using protein-ligand interaction fingerprints. Clusters derived from the interface grid revealed strong interactions with residues critical for YAP-TEAD binding. Among the 4 isomers, Ia-2 consistently showed the most favorable binding free energies. Notably, Cluster 7 highlighted a unique Ia-2 binding mode involving simultaneous interactions with Met86 and Arg87, suggesting a competitive mechanism at the YAP-TEAD interface. These results suggest that structural chirality may influence binding stability and interaction patterns, and that the Ia-2 isomer is predicted to preferentially stabilize an inhibitory binding mode. This study provides the first systematic comparison of all VP isomers with full-length YAP and suggests that isolating Ia-2 from Visudyne may enhance anticancer efficacy. The findings further support the rational strategies for designing selective YAP-TEAD inhibitors.

ROS-sensitive nanocarriers for synergistic X-PDT/chemo/immunotherapy of triple-negative breast cancer and metastasis.

Skin fibrosis results from excessive extracellular matrix (ECM) deposition and tissue remodeling due to persistent inflammation and mechanotransduction dysregulation. Current in vivo animal models lack human relevance, while conventional 2D and 3D in vitro models misrepresent physiological mechanical forces. To address this gap, we developed a miniaturized edgeless-skin chip (ESC) platform with gravity-driven perfusion, enabling enhanced biomechanical mimicry for fibrosis modeling. ESCs present bioengineered skin grown around a 3D-printed scaffold, mimicking the continuous geometry of human skin and in vivo mechanical balance. Compared to conventional skin constructs (CSCs) that have open boundaries on all sides, ESCs exhibited higher sensitivity to TGF-β1, leading to increased ECM deposition, myofibroblast activation, YAP signaling upregulation, matrix stiffness and reduced hydraulic permeability. Inhibiting YAP signaling with verteporfin (VTP) reduced collagen deposition, prevented tissue stiffening, and attenuated several fibrosis markers, confirming the role of mechanotransduction in fibrosis progression using human cells. Transcriptome analysis revealed upregulation of fibrosis-associated genes, including COL10A1, COL11A1, and ACTA2, counterbalanced by elevation of anti-fibrotic regulators such as DKK2, which suggests the activation of negative feedback mechanisms. These findings establish the ESC platform as a robust human-relevant mechanomimetic model for studying fibrosis and evaluating anti-fibrotic therapies, addressing a critical need for translational drug discovery.

Immunotherapy has emerged as a promising therapeutic modality for esophageal cancer (EC). Nevertheless, its clinical efficacy is frequently constrained by the overexpression of the vascular endothelial growth factor (VEGF) and the inherently low immunogenicity of EC cells. To address these limitations, we engineered a novel self-assembled nanoplatform utilizing the thin-film dispersion method. This platform comprises a biocompatible PVCL-PVA-PEG triblock copolymer designed for the codelivery of paclitaxel (PTX) and verteporfin (VER). This copolymer demonstrates a relatively low critical micelle concentration (CMC), thereby enhancing its stability and retention within physiological environments─attributes conferring advantages over conventional carriers, such as liposomes or PLGA-based nanoparticles. This study introduces a VER-based delivery system exhibiting a unique photoactivatable dual-function capability. In the absence of photostimulation (light-off state), nonphotoactivated VER acts as a potent Yes-associated protein (YAP) inhibitor, effectively attenuating VEGF-induced angiogenesis. Concurrently, PTX induces immunogenic cell death (ICD), activating antitumor immune responses. Upon photoirradiation (light-on state), VER-mediated photodynamic therapy (PDT) generates cytotoxic reactive oxygen species (ROS), directly eliminating tumor cells and further amplifying ICD. By integrating this spatiotemporally controlled dual-action mechanism of VER with PTX-induced chemotherapy, the nanoplatform significantly enhances the infiltration of tumoricidal immune effector cells. Consequently, this synergistic therapeutic strategy markedly improves the efficacy of chemo-photodynamic immunotherapy, offering a rational and promising treatment approach for esophageal cancer.

Fibrosis is characterized by excessive extracellular matrix (ECM) deposition driven by mechanical stress, yet the underlying molecular mechanisms remain incompletely understood. To investigate whether mechanical stress–induced ECM remodeling is mediated by YAP/TAZ–TEAD signaling and whether pharmacological modulation can mimic or reverse these effects. Nucleus pulposus (NP) cells were exposed to mechanical stress and treated with the MST1/2 inhibitor XMU-MP-1 or the YAP/TAZ–TEAD inhibitor Verteporfin (VP). Nuclear localization of YAP/TAZ–TEAD, expression of canonical TEAD target genes (CTGF, CYR61, ANKRD1), and ECM markers (Col1, Col2, α-SMA, FN, CTGF) were analyzed by Western blot, qPCR, and immunofluorescence. Mechanical stress induced nuclear accumulation of YAP/TAZ–TEAD, upregulated TEAD target genes, and promoted profibrotic ECM remodeling. XMU-MP-1 recapitulated these effects, while VP suppressed TEAD-dependent transcription and reversed ECM remodeling without affecting nuclear localization. YAP/TAZ–TEAD signaling mediates mechanically induced ECM remodeling. Activation of this pathway drives profibrotic gene expression, and disruption of YAP–TEAD interaction effectively reverses these changes, highlighting a potential therapeutic target for fibrosis.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-28960-0.

P-glycoprotein (P-gp, ABCB1)-mediated multidrug resistance (MDR) remains a significant barrier to successful chemotherapy outcomes for cancer patients. While photoactivation of verteporfin (VP), a photosensitizer, has demonstrated success for overcoming MDR through direct protein aggregation upon photoactivation and through adenosine triphosphate (ATP) depletion, the impact of VP's formulation on P-gp function and cellular energetics has not been fully characterized in this context. In this study, we screened four well-established VP formulations-liposomal VP (L-VP), lysophosphatidylcholine-conjugated VP (lysoPC VP), liposomal formulation of lysoPC VP (L-lysoPC VP), and a self-assembled VP nanoaggregate (NanoVP), with a free form of VP as a control-for their ability to inhibit P-gp. Using a combination of invitro intracellular VP accumulation assays, P-gp substrate retention experiments, and Seahorse-based metabolic profiling, we identified NanoVP as the lead formulation for P-gp modulation in cancer cells. NanoVP effectively depleted ATP in drug-resistant cancer cells, while being recognized as a P-gp substrate. Photodynamic priming with NanoVP at sub-cytotoxic light doses enhanced P-gp substrate retention within the cells without damaging P-gp protein, indicating ATP depletion as the primary mode of functional inhibition. These findings highlighted NanoVP's clinical potential to enhance chemotherapeutic efficacy via photoactivation-based modulation of P-gp's function in multidrug-resistant cancers.

Photodynamic therapy (PDT) is a highly selective, clinically approved, minimally invasive technique that effectively eliminates cancer cells. Its effectiveness is limited by poor light penetration into tissue and the hydrophobic nature of photosensitizers, highlighting the need for new approaches to treatment. Here, a theranostic upconversion nanoplatform, consisting of a NaYF4:Yb,Er,Tm,Fe core and a NaHoF4 shell codoped with Yb, Nd, Gd and Tb ions, was designed to enhance PDT outcomes by integrating multi-wavelength upconversion luminescence, T2-weighted magnetic resonance imaging (MRI) and PDT. The synthesized core–shell upconversion nanoparticles (CS-UCNPs) were coated with new verteporfin (VP)-conjugated alendronate-terminated poly(N,N-dimethylacrylamide-co-2-aminoethyl acrylate) [Ale-P(DMA-AEA)] grafted with poly(ethylene glycol) (PEG). Under 980 nm NIR irradiation, CS-UCNP@Ale-P(DMA-AEA)-PEG-VP nanoparticles generated reactive oxygen species (ROS) due to the efficient energy transfer between CS-UCNPs and VP. In a pilot preclinical study, intratumoral administration of nanoparticle conjugates to mice, followed by exposure to NIR light, induced necrosis of pancreatic tumor and suppressed its growth.

Verteporfin is a broad-spectrum inhibitor of arboviruses and influences viral and host-based events.

Targeting YAP1-dependent aerobic glycolysis to mitigate fibrosis progression in morphea.

Abstract BACKGROUND Glioblastoma multiforme (GBM) is one of the most aggressive and treatment-resistant forms of brain cancer. Current therapeutic strategies, including surgery, chemotherapy, and radiotherapy, often fail due to the tumor’s ability to develop resistance. The proteins YAP-1 (Yes-associated protein 1) and PARP-1 (Poly-(ADP-ribose)-polymerase-1) have been implicated in this resistance, playing crucial roles in cell proliferation and DNA repair mechanisms, respectively. MATERIAL AND METHODS This study explored the inhibitory potential of natural compounds from Lepidium meyenii (Peruvian Maca) on the YAP-1 and PARP-1 protein systems to develop novel therapeutic strategies for GBM. By molecular dynamics simulations, we identified N-(3-Methoxybenzyl)-(9Z,12Z,15Z)-octadecatrienamide (DK5) as the most promising natural inhibitor for PARP-1 and stearic acid (GK4) for YAP-1 RESULTS Although synthetic inhibitors, such as Olaparib (ODK) for PARP-1 and Verteporfin (VER) for YAP-1, only VER was superior to the naturally occurring molecule and proved a promising alternative. CONCLUSION In conclusion, natural compounds from Lepidium meyenii (Peruvian Maca) offer a potentially innovative approach to improve GBM treatment, complementing existing therapies with their inhibitory action on PARP-1 and YAP-1

IntroductionMechanical ventilation (MV) activates inflammatory signaling pathways, leading to ventilator-induced lung injury (VILI), the activation of lung repair processes, persistent inflammatory stimulation and incomplete tissue repair leads to pulmonary fibrosis. The role of Yes-associated protein (YAP) in VILI and related tissue repair mechanisms remains elusive. MethodsWe examined the effects of inhibiting or stimulating YAP activity on VILI, lung repair and fibrosis in a mouse model of MV-induced lung injury. Mice were subjected to either low tidal volume ventilation (LVT) or high tidal volume ventilation (HVT), and HVT was used in subsequent experiments. Additional mice were treated with or without the YAP inhibitor verteporfin (VP) and with or without the YAP stimulator XMU-MP-1 (X) and then subjected to HVT. The severity of lung injury and fibrosis was evaluated via histological analysis; the extent of lung repair was tested by measuring the levels of alveolar epithelial cell (AEC) marker proteins; YAP activity was assessed via Western blotting, immunoprecipitation and immunofluorescence. ResultsMV caused lung injury and fibrosis, decreased the protein expression of AEC markers and β-catenin, increased YAP expression, and the effect of HVT was greater than that of LVT. After inhibition of YAP activity, HVT decreased β-catenin expression, further inhibiting regeneration of AECs and worsening lung injury and fibrosis. In contrast, after stimulation of YAP activity, the reduction in β-catenin was mitigated, the impairment of AEC regeneration was ameliorated, lung injury and fibrosis were alleviated. DiscussionThe results indicate stimulation of YAP activity alleviates VILI by promoting lung repair and inhibiting fibrosis development.

In lung cancer, the tyrosine kinase receptor c-MET is often overexpressed, driving tumour progression and metastasis. This aberrant surface expression distinguishes the tumour cells from surrounding healthy tissue, providing an opportunity for targeted delivery of cytotoxic agents. This study aimed at designing and developing a tailor-made engineered nanoparticulate platform tuneable for the selective targeting of c-MET overexpressing cells. For this purpose, an effective conjugation method of a potent in-house developed single-chain variable fragment (3H3-HisC scFv) with PLGA-based nanoparticles (NPs) was set up for targeted delivery of the antitumor agent verteporfin (VP) to lung cancer cells (A549). The 3H3-HisC scFv was modified to allow site-directed sulfhydryl-reactive conjugation at the C-terminus, preserving its integrity and binding capacity. Comprehensive characterization confirmed the NP functionalization and drug-loading efficiency (3µg VP/mg NPs). When tested in vitro, the nanoplatform demonstrated specific enhanced uptake into c-MET-overexpressing A549 cells after 1h of incubation. A key advantage of this nanoplatform is its flexibility in modulating the VP release rate by adjusting the structural polymer's Mw or L:G ratio, without altering the functionalization, allowing for application-specific customization.

Overcoming hypoxia and enhancing therapeutic precision remain critical challenges for sonodynamic therapy (SDT) in oncology. Herein, we develop a biomimetic liposomal platform (DiR-VT@cmLipo) coencapsulating the sonosensitizer verteporfin (VP) and hypoxia-activated prodrug evofosfamide (TH302), which synergistically inhibits tumor progression via fluorescence imaging-guided ultrasound-activated spatiotemporally selective sonodynamic-chemotherapy. Engineered with natural membrane components, DiR-VT@cmLipo exhibits prolonged systemic circulation while maintaining precise tumor-specific accumulation after intravenous injection. The therapeutic cascade was precisely initiated through an ultrasound-triggered VP-mediated ROS burst, simultaneously consuming intratumoral oxygen. This creates a self-amplifying hypoxia gradient to promote the activation of cytotoxic payload TH302, enhancing SDT efficacy through synergistic mechanisms. This biomimetic nanoplatform represents an innovative strategy for overcoming microenvironmental limitations in SDT, establishing a paradigm for synergistic tumor microenvironment remodeling and precision-controlled combination therapy. The cascaded self-amplifying activation mechanism and spatiotemporally tumor-selective therapeutic amplification position DiR-VT@cmLipo as a promising candidate for clinical translation in solid tumor management.

Oxidative stress activates YAP/TEAD1/NCOA4 axis to promote ferroptosis of endplate chondrocytes and aggravate intervertebral disc degeneration.