ArticleJournal of nanobiotechnology2026
In silico design of CLDN4-directed peptide-loaded nanobubbles for ultrasound-assisted delivery, photothermal therapy, immune activation, and multimodal imaging in ovarian cancer.
Article in Journal of nanobiotechnology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundOvarian cancer remains among the most lethal gynecologic malignancies, partly because occult peritoneal disease limits complete macroscopic cytoreduction and image-guided intervention. Claudin 4 (CLDN4) is enriched in epithelial ovarian carcinoma but is not tumor-exclusive; therefore, rational CLDN4-directed delivery requires consideration of tumor enrichment, surface accessibility, and ligand engagement. This study aimed to develop a CLDN4-directed peptide-nanobubble phototheranostic platform integrating molecular recognition, ultrasound/fluorescence imaging, ultrasound-associated IP-related delivery/retention, and photothermal treatment.
resultsStructure-guided in silico optimization and surface plasmon resonance validation identified P15 as a CLDN4-binding peptide with nanomolar affinity (steady-state K_D = 11.28 nM). P15 was conjugated with indocyanine green (ICG) to generate an ICG-P15 photothermal conjugate (IP), which was incorporated into sulfur hexafluoride-filled lipid nanobubbles to form IP/NBs. Surface-preserving binding and competition assays, non-permeabilized CLDN4 immunofluorescence, cellular uptake imaging, and patient-derived organoid assays supported CLDN4-associated P15/IP interaction and intratissue penetration in the tested ovarian cancer models. IP/NBs preserved ultrasound visibility and photothermal responsiveness and showed ultrasound-associated changes in IP release and transport-related readouts under the applied acoustic conditions. In vivo imaging demonstrated tumor-associated IP-related fluorescence after IP/NBs administration, with D-NBs serving as a non-targeted dye-loaded nanobubble comparator. Matched post-ultrasound imaging further showed higher tumor-associated fluorescence in the IP/NBs + ultrasound group than in the no-ultrasound group, and an IP/NBs + near-infrared irradiation group without ultrasound showed intermediate tumor suppression compared with the full IP/NBs + ultrasound + near-infrared treatment. The full treatment regimen produced marked tumor suppression and was associated with increased oxidative-stress readouts, calreticulin exposure, ATP/HMGB1 release, cytokine secretion, immune-cell activation, and tumor-associated T-cell infiltration.
conclusionsThis study develops and evaluates a CLDN4-directed, IP-containing nanobubble framework for ovarian cancer phototheranostics. The data support ultrasound-associated enhancement of tumor IP-related delivery/retention, photothermal tumor suppression, and treatment-associated immune activation under the tested workflow, while defining areas for further optimization of peptide stability, nanobubble pharmacology, acoustic dosing, and translational safety.
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