ACTA PHARMACOLOGICA SINICA
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2025
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IF 8.4
Lu Bin,Sun Yi-yun,Zhou Jia-hao,Chen Dan-ni,Guo Yue,Chen Yu-lu,Pan Cheng-hao,Chen Zheng-yang,He Qiao-jun,Yuan Meng,Cao Ji,Jiang Li,Yang Bo
Ubiquitin-specific protease 14 (USP14) is a crucial modulator of proteasomal function and cellular proteostasis, which plays an important role in the development and progression of various cancers including colorectal cancer (CRC). In this study we screened 670 covalent compounds using the in vitro Ub-AMC hydrolysis assay, and identified AKOS, initially a Chikungunya virus inhibitor, as a novel small-molecule inhibitor of USP14. We showed that AKOS inhibiting USP14 deubiquitinase activity with an IC 50 value of 0.98 μM. AKOS directly bound to USP14, covalently modifying the active-site cysteine residue (Cys 114 ), thereby effectively inhibiting its deubiquitinase activity. We demonstrated that inhibition of USP14 by AKOS might destabilize MEF2D, a critical substrate, resulting in downregulation of the expression and translation of ECM-related transcription factors such as ITGB4 . AKOS exhibited potent anti-cancer effects: the USP14 inhibitor significantly inhibited the proliferation and metastasis of CRC cells in vitro with IC 50 values of 9.88 and 16.57 μM, respectively, in SW620 cells and HCT116 cells. Intratumoral injection of AKOS (15, 30 mg/kg, every 5 days) effectively suppressed the tumor growth in HCT116 xenograft mouse models in vivo. Collectively, we demonstrate that AKOS is a promising chemical probe for targeting USP14 in CRC, offering a novel strategy for disrupting the malignant progression of CRC.
研究领域:
癌症生物学
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生物化学
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病毒学
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药理学
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细胞生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
iScience
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2026
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IF 4.5
Dan Xiao,Jun Fang,Hui Jian,Yang Yu
Combining immunotherapy with other treatments improves survival in colorectal cancer, yet some patients remain unresponsive. Tumor-associated macrophages (TAMs) are a key immune cell population driving this immunotherapy resistance and fostering an immunosuppressive microenvironment. To overcome this, we screened a deubiquitinating enzyme (DUB) library targeting TAMs and identified USP14 as specifically upregulated in TAMs. Inhibiting USP14 reversed their pro-tumor functions, promoted M1 polarization, enhanced tumor cell killing, and activated effector T cells. USP14 inhibition also increased PD-L1 expression on tumor cells, alleviating T-cell suppression. In vivo , combining a USP14 inhibitor with an anti-PD-1 antibody synergistically enhanced immunotherapy efficacy, suppressed tumor progression, and improved survival in a mouse colon cancer model. Thus, USP14 is a promising target to overcome immunotherapy resistance in colorectal cancer.
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产品关联
DOI
高影响因子
Heliyon
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2024
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IF 3.4
Lina Ba,Mingyao E,Ruixuan Wang,Nan Wu,Rui Wang,Renling Liu,Xiang Feng,Hanping Qi,Hongli Sun,Guofen Qiao
Background Cardiac remodeling is a common pathological feature in many cardiac diseases, characterized by cardiac hypertrophy and fibrosis. Triptolide (TP) is a natural compound derived from Tripterygium wilfordii Hook F. However, the related mechanism of it in cardiac remodeling has not been fully understood. Methods and results Transverse aortic constriction (TAC)-induced cardiac hypertrophic mouse model and angiotensin II (Ang II)-induced cardiomyocytes hypertrophic model were performed. Firstly, the results indicate that TP can improve cardiac function, decreased cardiomyocyte surface area and fibrosis area, as well as lowered the protein expressions of brain natriuretic peptide (BNP), β-major histocompatibility complex (β-MHC), type I and III collagen (Col I and III). Secondly, TP suppressed cardiac pyroptosis, and decreased the levels of Interleukin-1β (IL-1β), Interleukin-18 (IL-18) by Enzyme-linked immunosorbent assay (ELISA), and pyroptosis-associated proteins. Furthermore, TP enhanced the expressions of Nuclear factor erythroid 2-related factor 2 (Nrf2) and Heme oxygenase 1 (HO-1). Interestingly, when Nrf2 was silenced by siRNA, TP lost its properties of reducing pyroptosis and cardiac hypertrophy. In addition, in the Transforming Growth Factor β1 (TGF-β1)-induced primary human coronary artery endothelial cells (HCAEC) model, TP was found to inhibit the process of endothelial-to-mesenchymal transition (EndMT), characterized by the loss of endothelial-specific markers and the gain of mesenchymal markers. This was accompanied by a suppression of Slug, Snail, and Twist expression. Meanwhile, the inhibitory effect of TP on EndMT was weakened when Nrf2 was silenced by siRNA. Lastly, potential targets of TP were identified through network pharmacology analysis, and found that Ubiquitin-Specific Protease 14 (USP14) was one of them. Simultaneously, the data indicated that decrease the upregulation of USP14 and Kelch-like ECH-Associated Protein 1 (Keap1) caused by cardiac remodeling. However, Keap1 was decreased and Nrf2 was increased when USP14 was silenced. Furthermore, CoIP analysis showed that USP14 directly interacts with Keap1. Conclusion TP can observably reduce pyroptosis and EndMT by targeting the USP14/Keap1/Nrf2 pathway, thereby significantly attenuating cardiac remodeling.
研究领域:
心脏病学
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药理学
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分子生物学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AFFECTIVE DISORDERS
·
2025
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IF 4.9
Huiying Xie,Yan Xue,Rui Deng,Liuxiang Chen,Yiyi Wang,Jia Xu,Hao Liu,Xingxing Li,Wei Cui,Jun-Fang Zhang
Background Stress is a significant trigger of depression, and miR-124 has been implicated in stress-induced depression, while the underlying molecular mechanisms remain unclear. Methods We established mouse depression models using chronic restraint stress (CRS) and chronic unpredictable mild stress (CUMS), and the HT-22 cell models with corticosterone (CORT) and glutamate (Glu). Depression-like behaviors were assessed using the forced swimming test, tail suspension test, and sucrose preference test. Adeno-associated viruses were employed to manipulate miR-124 and ubiquitin-specific protease 14 (USP14) expression. mRNA levels of miR-124 , Usp14 , and Rest were measured by qRT-PCR, the interaction between miR-124 and Usp14 mRNA was assessed by luciferase reporter assays, protein expression and interaction of USP14 and repressor element 1 silencing transcription factor (REST) were detected by western blotting, ubiquitination assay and co-immunoprecipitation. Immunofluorescence visualized altered REST expression in hippocampal neurons. Results CRS and CUMS robustly increased miR-124 levels in the hippocampus, while decreasing the USP14 and REST expression. Similar expression changes were seen in CORT- and Glu-treated HT-22 cells. Mice with miR-124 overexpression but not inhibition in the hippocampus exhibited notable depression-like behavior. Mechanistically, miR-124 directly targeted USP14, reduced USP14 binding to REST, thereby increasing REST ubiquitination and degradation. Inhibition of miR-124 produced antidepressant-like effects and restored the expression and interaction of USP14 and REST in the CRS and CUMS mice. Furthermore, overexpression of USP14 alleviated depression-like behaviors and low REST levels. Conclusion Hippocampal miR-124 mediates depression-like behavior under chronic stress via regulating the USP14/REST signaling pathway, highlighting its potential as a therapeutic target.
研究领域:
神经生物学
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分子精神病学
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信号转导
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表观遗传学
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应激生理学
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神经药理学
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产品关联
DOI
PubMed
高影响因子
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