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Identification and Functional Analysis of Targets of Dehydrodiisoeugenol in Bladder Cancer Based on
Zhao Zhai1, Fan Wu2, Guoli Sheng2
1Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Urology, Peking University Cancer Hospital & Institute, Beijing 100089, China.
Abstract:
Background/Objectives: The clinical management of bladder cancer is severely impeded by high recurrence rates and the rapid emergence of chemoresistance, necessitating the discovery of novel therapeutic agents with distinct mechanisms of action. Dehydrodiisoeugenol (DHE), a bioactive neolignan, exhibits potent anti-tumor efficacy, yet its direct molecular targets and mode of action remain elusive. Methods: To deconvolute the mechanism of DHE, we integrated a phenotypic screening approach using 2D cell lines and 3D patient-derived organoids with a chemoproteomics-based activity-based protein profiling (ABPP) strategy. We synthesized a functionalized photoaffinity probe to capture the specific interactome of DHE under physiological conditions and validated targets via cellular thermal shift assays (CETSA), quantitative mass spectrometry, and 100 ns molecular dynamics (MD) simulations. Results: DHE exhibited potent dose-dependent cytotoxicity in bladder cancer cells, with IC50 values of 39.23 μM in T24 and 34.58 μM in 5637 cells. In 3D patient-derived organoids, DHE significantly reduced viability (p < 0.0001). Using a dual-filtering ABPP strategy, we identified 65 high-confidence candidate targets, prioritizing PTPN1 (PTP1B) as the primary functional interactor. Comparative molecular docking and 100 ns MD analyses showed that multiple stereoisomers of DHE could adopt plausible PTPN1-binding modes. Mechanistically, organoid proteomics indicated that DHE engagement with PTPN1 disrupts ER membrane homeostasis, thereby modulating the PI3K-Akt signaling axes. Conclusions: These findings establish PTPN1 as a critical druggable vulnerability in bladder cancer and define the molecular basis for the therapeutic potential of DHE. This study highlights the power of combining chemoproteomics with physiological 3D models to accelerate the translation of natural products into precision cancer therapies.
Insights
Dehydrodiisoeugenol (DHE) shows promise in treating bladder cancer by targeting PTPN1, a key protein involved in cell signaling and homeostasis. This natural compound effectively reduces tumor cell viability, offering a new therapeutic avenue.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Bladder cancer management is challenged by high recurrence and chemoresistance.
- Novel therapeutic agents with distinct mechanisms are needed.
- The molecular targets and action of Dehydrodiisoeugenol (DHE) were previously unknown.
Purpose of the Study:
- To elucidate the mechanism of action of Dehydrodiisoeugenol (DHE) in bladder cancer.
- To identify direct molecular targets of DHE.
- To explore DHE's therapeutic potential.
Main Methods:
- Integrated phenotypic screening with 2D cell lines and 3D patient-derived organoids.
- Employed chemoproteomics-based activity-based protein profiling (ABPP) with a photoaffinity probe.
- Validated targets using CETSA, mass spectrometry, and molecular dynamics (MD) simulations.
Main Results:
- DHE demonstrated potent dose-dependent cytotoxicity against bladder cancer cells (T24, 5637) and reduced viability in 3D organoids.
- Identified 65 high-confidence candidate targets, with PTPN1 (PTP1B) prioritized as a key interactor.
- DHE's engagement with PTPN1 disrupts ER membrane homeostasis, modulating PI3K-Akt signaling.
Conclusions:
- PTPN1 is a critical druggable target in bladder cancer.
- The study defines the molecular basis for DHE's therapeutic potential.
- Combined chemoproteomics and 3D models accelerate natural product translation for precision cancer therapy.
