Wound healing involving persistent inflammation and fibrosis is driven by reactive oxygen species (ROS) and Yes-associated protein (YAP), often leading to scars. A polyvinyl alcohol/alginate ultrasound-responsive hydrogel was developed, containing free epigallocatechin gallate (EGCG) in the matrix and EGCG-verteporfin (VP) nanoparticles embedded in the hydrogel network. Free EGCG is released rapidly, whereas EGCG-VP nanoparticles release is triggered by ultrasound. In vitro, this system reduced ROS by ∼69%. Western blot revealed ∼71% reduction in α-smooth muscle actin and ∼47% in YAP. In vivo, it promoted wound closure in rats. This system enables programmable modulation of the wound microenvironment.

Tendon healing is frequently accompanied by fibrosis, scar formation and incomplete functional recovery. Tendon stem cells (TSCs) are central to tendon regeneration; however, the molecular basis by which low-intensity pulsed ultrasound (LIPUS) regulates their tenogenesis-associated behavior through mechanotransduction remains incompletely understood. In the present study, it was investigated whether LIPUS enhances tenogenesis-associated responses and matrix-related gene expression in rat TSCs through the YAP/TAZ/SCX signaling axis. Public transcriptomic data from the Gene Expression Omnibus database (GSE273466) were analyzed to identify differentially expressed genes and enriched pathways associated with LIPUS exposure. Primary rat Achilles tendon-derived TSCs were isolated, expanded and characterized by morphology and flow cytometry. Passage-3 TSCs were assigned to the Control, LIPUS and LIPUS + Verteporfin (VP) groups. Cell migratory behavior was evaluated using a scratch assay. Immunofluorescence staining, western blotting and reverse transcription-quantitative PCR were performed to assess the expression of YAP, TAZ, SCX, COL I and TNMD. Transcriptomic analysis indicated that LIPUS enriched mechanotransduction-related, cytoskeleton-associated, and YAP/TAZ-related signaling programs, together with enhanced SCX-associated signatures. Cultured cells exhibited a typical TSC phenotype, characterized by positive expression of CD90, CD105 and CD146 and negative expression of CD11b, CD34 and CD45. LIPUS increased scratch closure at 24 and 48 h and upregulated YAP, TAZ, SCX, COL I and TNMD at the protein and/or mRNA level. These effects were attenuated by VP treatment. In conclusion, LIPUS enhances a tenogenesis-associated molecular phenotype in TSCs and promotes matrix-related gene expression, at least in part through mechanotransduction-dependent activation of YAP/TAZ signaling and upregulation of SCX. These findings provide mechanistic support for further investigation of LIPUS as a strategy for tendon regeneration.

The current study aims to present a novel approach to alleviate the aggressiveness of ovarian cancer by pharmacological disruption of the Yes-Associated Protein (YAP) - 1/Transcriptional Enhancer Factor domain (TEAD) heterodimer structure using Carmofur. The Ovcar-3 cell line was incubated with progressively diluted Carmofur solutions. Proliferation inhibition was assessed using the MTT assay. Flow cytometry was employed to determine the pro-apoptotic properties. Cellular morphological alterations were assessed with confocal microscopy. The migration inhibition status was assessed via the scratch assay, whilst the colony formation inhibition level was evaluated using the colony assay. The western blotting technique was employed to determine YAP-1 expression levels. The disruptive impact of Ceramide and Carmofur on the YAP-1/TEAD heterodimer structure was elucidated via molecular modeling, considering three interfaces: the β-strand, the α-helix, and the Ω-loop. Furthermore, validation was conducted by assessing the disruptive impact of Verteporfin on the YAP-1/TEAD heterodimer. The findings indicated that Carmofur diminished cell viability in a concentration- and time-dependent manner, with IC50 values of 17.26 µM at 24 h, 10.15 µM at 48 h, and 9.22 µM at 72 h, respectively. Carmofur showed a 26.65% apoptotic effect at the IC50 value over 24 h. Additionally, Carmofur diminished the relative YAP-1 levels from 9.72 to 2.98 in the Ovcar-3 cell line and exhibited an anti-metastatic effect by suppressing colony formation and migratory capabilities. This is the first study to provide in silico and in vitro evidence that Carmofur induces apoptosis by disrupting the YAP-1/TEAD heterodimer, thereby inhibiting the proliferation and migration of Ovcar-3 cells.

Photodynamic therapy (PDT) is a clinically established photochemical treatment in which light activation of photosensitizers (PS) generates reactive species, such as reactive oxygen species (ROS), that induce localized cytotoxicity. Beyond direct tumor ablation, PDT can modulate the tumor microenvironment, enhancing antitumor immunity and sensitizing tumors to combination therapies. Related sublethal regimens, termed photodynamic priming (PDP), aim to exploit these immunomodulatory effects. While deep tumors can be accessed using fiber-optic or endoscopic light delivery, extending PDT or PDP to disseminated metastatic disease would require numerous invasive insertions, limiting clinical practicality. Here, we examine immune cell-mediated light delivery, in which luciferase-expressing immune cells infiltrate tumors and act as internal bioluminescent sources for photodynamic activation. Although this strategy could enable PDT and PDP across widespread lesions without external illumination, it remains unclear whether far-field photon emission from luciferase is sufficient to activate PS, or whether near-field mechanisms such as bioluminescence resonance energy transfer are required. Using luciferase-engineered tumor-associated macrophages and verteporfin, we observed no detectable ROS generation or tumor cell cytotoxicity in vitro. Complementary Monte Carlo simulations incorporating tissue scattering and absorption show that achievable photon fluxes are several orders of magnitude below established thresholds for cytotoxic PDT. These findings quantitatively define the limits of far-field bioluminescence and highlight the need for near-field strategies for immune cell mediated PDT and PDP.

The main causes of posterior capsular opacification (PCO) are the proliferation, migration, and epithelial-mesenchymal transition (EMT) of leftover lens epithelial cells (LECs), though the exact mechanisms of EMT in PCO are not completely known. This research sought to determine whether PCO progression depends on Yes-associated protein 1 (YAP1) activation and EMT in LECs. We conducted gain- and loss-of-function experiments in human lens epithelial cells (HLE-B3) using YAP1-overexpressing plasmids and siRNA-YAP1. Cell migration was tested with a scratch wound assay, and Hippo/YAP signaling and EMT markers were analyzed via qPCR, Western blot, and immunocytochemistry. HLE-B3 cells were exposed to transforming growth factor-beta 2 (TGF-β2) and the YAP1 inhibitor verteporfin (VP) to evaluate cell proliferation, migration, and molecular expression using CCK-8, scratch assay, qPCR, and Western blot. A rabbit model for PCO was developed to test VP's effectiveness, evaluated by slit-lamp photography, H&E staining, immunofluorescence, and Western blotting for YAP1 and EMT-related proteins. In vitro, YAP1 overexpression increased TEAD2 and BIRC5, promoting EMT, while YAP1 knockdown reduced these proteins, inhibiting EMT. VP treatment decreased HLE-B3 proliferation and migration. Inhibiting YAP1 and its downstream targets, CTGF and BIRC5, suppressed EMT. In vivo, VP significantly delayed PCO progression in rabbits and decreased YAP1, α-SMA, and vimentin expression. These results suggest that YAP1 facilitates EMT in lens epithelial cells through the Hippo/YAP pathway, and its inhibition could be a promising strategy for PCO treatment.

Verteporfin (VP) is widely used as a light-independent pharmacologic inhibitor of the YAP-TEAD transcriptional complex, despite its potent photosensitizing properties. Here we show that VP-associated high-molecular-weight complexes and apparent depletion of key proteins on immunoblots can be substantially driven by ambient-light exposure during post-lysis handling, more than by light during cellular treatment. In this study, the hypothesis is tested independently using MEL270, HEK293, and MCF-7 cells treated with therapeutic and supra-therapeutic concentrations of VP under four systematically varied light/dark conditions. The results demonstrate that high-molecular-weight complex (HMWC) formation for p62, DIAP1, ROCK1, YAP, and phospho-YAP is driven principally by light exposure during sample processing rather than during cell treatment, occurs in cell-free lysates within 1 hour of ambient-light exposure, and is markedly attenuated by both N-acetylcysteine and L-histidine, consistent with contributions from both radical-mediated and singlet oxygen-mediated photochemistry. Notably, YAP and phospho-YAP are highly susceptible to photo-cross-linking, whereas transcription enhancer factor 1 (TEF1)/TEAD1 remains comparatively resistant, a pattern that mimics selective YAP-TEAD inhibition on standard immunoblots. In a single-experiment CCK-8 assay, viability was markedly lower under light than in darkness at 24 hours, an observation consistent with the immunoblot findings but requiring independent replication. These findings support the light-artifact hypothesis and may have implications for how preclinical data on VP are interpreted in the design of clinical investigations.

Subthreshold micropulse laser (SML) is a phototherapeutic strategy for central serous chorioretinopathy (CSC) and has been reported for selected serous pigment epithelial detachments (PEDs), especially when the verteporfin in photodynamic therapy (PDT) is unavailable. However, retinal pigment epithelium (RPE) tear following SML has not been well documented. This case report describes an RPE tear and chronic CSC-like subretinal fluid after SML treatment for serous PED in a 44-year-old male. Staged SML and focal laser photocoagulation guided by sequential multimodal imaging completely resolved the refractory subretinal fluid, with stable visual function and no recurrence during long-term follow-up. This report highlights a potential safety risk of SML for serous PED, underscoring the value of multimodal imaging in guiding treatment with sequential SML or focal photocoagulation for chronic CSC-like fluid.

While anti-PD-L1 antibody (αPD-L1) therapy holds promise, its efficacy against lung cancer brain metastasis (LCBM) is severely limited by the blood-brain barrier (BBB), the immunosuppressive tumor microenvironment and adaptive immune resistance. To overcome these barriers, we engineered a pH-responsive nanocomposite (VP-αPD-L1@REB) by functionalizing brain-metastatic tumor cell-derived exosomes (EB) with RGD peptides (REB) for targeted co-delivery of verteporfin (VP) and αPD-L1. Benefiting from homotypic affinity and integrin-mediated transcytosis, VP-αPD-L1@REB efficiently crosses the BBB, accumulates within intracranial tumors, and undergoes pH-responsive cargo release. Mechanistically, VP induces a lethal reactive oxygen species (ROS) storm for direct tumor ablation. Simultaneously, VP downregulates the chaperone protein CMTM6 and activates cellular autophagy, forcibly driving internalized PD-L1 toward degradation via dual "endosome-lysosome" and "autophagy-lysosome" pathways. Driven by the synergy of VP's robust intracellular clearance and αPD-L1's surface blockade, this targeted nanoplatform successfully remodels the intracranial immunosuppressive microenvironment and triggers potent systemic anti-tumor immunity. This study provides a highly promising translational paradigm for overcoming adaptive immune resistance in central nervous system (CNS) malignancies.

Idiopathic pulmonary fibrosis (IPF) is driven by persistent fibroblast activation, excessive extracellular matrix (ECM) deposition, and progressive tissue stiffening, yet the mechanosensitive signaling programs that sustain fibrotic fibroblast states remain incompletely defined. Here, we investigated the association between a periostin (POSTN)-enriched fibrotic program and PIEZO1-dependent Yes-associated protein (YAP)/TEA domain transcription factor (TEAD) activation in IPF. Integrated analysis of public bulk lung transcriptomic datasets identified a reproducible POSTN-enriched ECM remodeling program in IPF lungs. Single-cell and donor-level pseudobulk analyses further showed that a PIEZO1/YAP activity-related signature was co-enriched in POSTN-positive fibrotic fibroblast and myofibroblast compartments. In bleomycin-induced pulmonary fibrosis, Postn upregulation, Piezo1 induction, and increased Yap nuclear localization were recapitulated in fibrotic lung regions. Verteporfin, used as a pharmacological inhibitor of Yap/TEAD-associated transcription, attenuated collagen deposition, hydroxyproline accumulation, profibrotic gene expression, and Yap target protein induction in vivo. In transforming growth factor-β1 (TGF-β1)-stimulated lung fibroblasts, verteporfin suppressed fibroblast migration, Yap nuclear localization, and profibrotic marker expression at non-cytotoxic concentrations. Knockdown of Piezo1 or Yap1 reduced TGF-β1-induced fibroblast activation, while Piezo1 knockdown decreased Yap nuclear localization and Yap/TEAD reporter activity. Active YAP partially restored profibrotic gene expression after Piezo1 knockdown, supporting Yap/TEAD as an important downstream effector of Piezo1. Collectively, these findings identify an association between a POSTN-enriched fibrotic fibroblast state and enhanced PIEZO1-YAP/TEAD signaling, while functional experiments support a role for PIEZO1-YAP/TEAD in fibroblast activation and fibrotic lung remodeling.

Cabazitaxel (CTX) exhibits potent antineoplastic activity and is demonstrated to promote antitumor immunity by triggering immunogenic cell death (ICD). This is commonly used in the clinical treatment of advanced and metastatic triple-negative breast cancer (TNBC). However, the therapeutic index of commercial CTX injections is severely restricted by their definite systemic toxicity and low tolerability. Herein, we developed biomimetic low-density lipoprotein nanoparticles (CTX2TK&VP@LDL NPs) by integrating ROS-responsive CTX dimeric prodrugs (CTX2TK) and the photosensitizer verteporfin (VP) into LDL NPs to selectively target tumors and achieve synergistic chemophotodynamic therapy and immunotherapy for TNBC with improved biosafety. CTX2TK&VP@LDL NPs were able to enhance the accumulation and cellular uptake in 4T1 tumors overexpressing LDL receptor, improving the targeted codelivery efficiency. When CTX2TK&VP@LDL NPs were deposited at the tumor sites, localized 1O2 was generated upon irradiation for triggering ultrafast CTX release with low premature CTX leakage in the off-target tissue. Additionally, the generated 1O2 can further synergistically reinforce ICD and elicit robust antitumor immune activation. The combination treatment with CTX2TK&VP@LDL NPs and PD-L1 blockade notably inhibited abscopal tumor growth and lung metastasis. Taken together, the biomimetic CTX2TK&VP@LDL NPs exploit endogenous LDL functionality to enable receptor-mediated tumor targeting and synergistic chemophotodynamic immune activation, providing an effective and safe therapeutic strategy for TNBC.

G protein-coupled receptor 87 (GPR87) has been implicated in the growth and progression of several malignancies, but its role in cutaneous squamous cell carcinoma (cSCC) remains poorly understood. Here, we found that GPR87 was aberrantly upregulated in murine and human skin tumors and in 12-O-tetradecanoylphorbol-13-acetate (TPA)-stimulated HaCaT keratinocytes. Using a 7,12-dimethylbenz[a]anthracene (DMBA)/TPA-induced mouse skin tumor model, we demonstrated that GPR87 knockout significantly delayed epidermal hyperplasia, tumor initiation, and malignant progression. Consistently, GPR87 knockdown suppresses TPA-induced keratinocyte proliferation, migration, stemness, and epithelial-mesenchymal transition in keratinocytes and cSCC cells. Mechanistically, GPR87 deficiency markedly reduced SOX2 expression in cultured cells and skin lesions. GPR87 knockdown also enhanced mammalian STE20-like serine/threonine kinases 1/2 (MST1/2) and large tumor suppressor kinases (LATS1/2) phosphorylation, increased inhibitory YAP phosphorylation, and suppressed Yes-associated protein (YAP) nuclear localization and downstream transcriptional activity. Importantly, activation of YAP by either YAP(S127A) expression or XMU-MP-1 treatment restored SOX2 expression in GPR87-depleted cells, whereas verteporfin-mediated inhibition of YAP-TEA domain transcription factor (TEAD) activity attenuated TPA-induced SOX2 upregulation. Together, these findings identify GPR87 as a key positive regulator of skin tumor initiation and progression and demonstrate that its tumor-promoting effects are mediated, at least in part, through the YAP-SOX2 axis. GPR87 may therefore represent a potential therapeutic target for cSCC.

Despite advances in medical technology, cancer treatment remains challenging. Our research focused on combining microtubule polymerization inhibitors with sonodynamic therapy (SDT) as a promising approach for cancer. SDT is a minimally invasive modality that uses tumor-selective sonosensitizers to induce the death of malignant and tumor-associated cells within the tumor microenvironment through targeted ultrasound irradiation. In this study, verteporfin was selected because of its dual properties of high ultrasound sensitivity and the ability to inhibit the yes-associated protein. Liposome-encapsulated docetaxel and verteporfin were employed to enhance tumor accumulation. This intervention demonstrated remarkable antitumor efficacy in three patients with ovarian metastases from gastric cancer, advanced gastric cancer, and lung metastases from breast cancer, respectively, without notable adverse events. Although the patients with ovarian and lung metastases had experienced progressive disease despite prior chemotherapy and immune checkpoint inhibitor treatment, our intervention successfully inhibited tumor progression. Furthermore, although conventional treatment options were considered unsuitable because of advanced age in the patient with gastric cancer, this intervention demonstrated a favorable safety profile and potential therapeutic benefit. To our knowledge, this study represents the first clinical report demonstrating the potential antitumor efficacy and safety of the combination of verteporfin liposomes, liposomal docetaxel, and SDT.