Cell Reports
·
2024
·
IF 7.5
Swati Keshri,Mariella Vicinanza,Michael Takla,David C. Rubinsztein
The transcription factor EB (TFEB) is a master regulator of lysosomal biogenesis and autophagy. We identify a distinct nuclear interactome of TFEB, with ubiquitin-specific protease 7 (USP7) emerging as a key post-translational modulator of TFEB. Genetic depletion and inhibition of USP7 reveal its critical role in preserving TFEB stability within both nuclear and cytoplasmic compartments. Specifically, USP7 is identified as the deubiquitinase responsible for removing the K48-linked polyubiquitination signal from TFEB at lysine residues K116, K264, and K274, thereby preventing its proteasomal degradation. Functional assays demonstrate the involvement of USP7 in preserving TFEB-mediated transcriptional responses to nutrient deprivation while also modulating autophagy flux and lysosome biogenesis. As USP7 is a deubiquitinase that protects TFEB from proteasomal degradation, these findings provide the foundation for therapeutic targeting of the USP7-TFEB axis in conditions characterized by TFEB dysregulation and metabolic abnormalities, particularly in certain cancers.
研究领域:
细胞生物学
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分子生物学
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医学
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DOI
PubMed
高影响因子
Autophagy
·
2024
·
IF 14.6
Zhigang Zhang,Shuai Chen,Shirui Jun,Xirong Xu,Yuchuan Hong,Xifei Yang,Liangyu Zou,You-Qiang Song,Yu Chen,Jie Tu
Individuals with genetic elimination of MLKL (mixed lineage kinase domain like pseudokinase) exhibit an increased susceptibility to neurodegenerative diseases like Alzheimer disease (AD). However, the mechanism is not yet fully understood. Here, we observed significant compromise in macroautophagy/autophagy in the brains of mlkl knockout (KO) mice, as evidenced by the downregulation of BECN1/Beclin1 and ULK1 (unc-51 like autophagy activating kinase 1). We identified UBA52 (ubiquitin A-52 residue ribosomal protein fusion product 1) as the binding partner of MLKL under physiological conditions. Loss of Mlkl induced a decrease in ubiquitin levels by preventing UBA52 cleavage. Furthermore, we demonstrated that the deubiquitinase (DUB) USP7 (ubiquitin specific peptidase 7) mediates the processing of UBA52, which is regulated by MLKL. Moreover, our results indicated that the reduction of BECN1 and ULK1 upon Mlkl loss is attributed to a decrease in their lysine 63 (K63)-linked polyubiquitination. Additionally, single-nucleus RNA sequencing revealed that the loss of Mlkl resulted in the disruption of multiple neurodegenerative disease-related pathways, including those associated with AD. These results were consistent with the observation of cognitive impairment in mlkl KO mice and exacerbation of AD pathologies in an AD mouse model with mlkl deletion. Taken together, our findings demonstrate that MLKL-USP7-UBA52 signaling is required for autophagy in brain through maintaining ubiquitin homeostasis, and highlight the contribution of Mlkl loss-induced ubiquitin deficits to the development of neurodegeneration. Thus, the maintenance of adequate levels of ubiquitin may provide a novel perspective to protect individuals from multiple neurodegenerative diseases through regulating autophagy.Abbreviations: 4HB: four-helix bundle; AAV: adeno-associated virus; AD: Alzheimer disease; AIF1: allograft inflammatory factor 1; APOE: apolipoprotein E; APP: amyloid beta precursor protein; Aβ: amyloid β; BECN1: beclin 1; co-IP: co-immunoprecipitation; DEGs: differentially expressed genes; DLG4: discs large MAGUK scaffold protein 4; DUB: deubiquitinase; EBSS: Earle’s balanced salt solution; GFAP: glial fibrillary acidic protein; HRP: horseradish peroxidase; IL1B: interleukin 1 beta; IL6: interleukin 6; IPed: immunoprecipitated; KEGG: Kyoto Encyclopedia of Genes and Genomes; KO: knockout; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MLKL: mixed lineage kinase domain like pseudokinase; NSA: necrosulfonamide; OPCs: oligodendrocyte precursor cells; PFA: paraformaldehyde; PsKD: pseudo-kinase domain; SYP: synaptophysin; UB: ubiquitin; UBA52: ubiquitin A-52 residue ribosomal protein fusion product 1; UCHL3: ubiquitin C-terminal hydrolase L3; ULK1: unc-51 like autophagy activating kinase 1; UMAP: uniform manifold approximation and projection; UPS: ubiquitin-proteasome system; USP7: ubiquitin specific peptidase 7; USP9X: ubiquitin specific peptidase 9 X-linked.
研究领域:
神经科学
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分子生物学
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细胞生物学
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遗传学
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DOI
高影响因子
BIOCHEMICAL PHARMACOLOGY
·
2023
·
IF 5.8
Xuepeng Wang,Chunchun Zou,Changju Hou,Zhenyu Bian,Wu Jiang,Maoqiang Li,Liulong Zhu
Mesenchymal stem cells (MSCs) and their derived extracellular vesicles (EVs) have emerged as promising tools for promoting bone regeneration. This study investigates the functions of EVs derived from bone marrow-derived MSCs (BMSCs) in osteoporosis (OP) and the molecular mechanism. EVs were isolated from primary BMSCs in mice. A mouse model with OP was induced by ovariectomy. Treatment with EVs restored bone mass and strength, attenuated trabecular bone loss and cartilage damage, and increased osteogenesis while suppressing osteoclastogenesis in ovariectomized mice. In vitro , the EVs treatment improved the osteogenic differentiation of MC-3T3 while inhibiting osteoclastic differentiation of RAW264.7 cells. Microarray analysis revealed a significant upregulation of ubiquitin specific peptidase 7 (USP7) expression in mouse bone tissues following EV treatment. USP7 was found to interact with Yes1 associated transcriptional regulator (YAP1) and stabilize YAP1 protein through deubiquitination modification. YAP1-related genes were enriched in the Wnt/β-catenin signaling, and overexpression of YAP1 promoted the nuclear translocation of β-catenin. Functional experiments underscored the critical role of maintaining USP7, YAP1, and β-catenin levels in the pro-osteogenic and anti-osteoclastogenic properties of the BMSC-EVs. In conclusion, this study demonstrates that USP7, delivered by BMSC-derived EVs, stabilizes YAP1 protein, thereby ameliorating bone formation in OP through the Wnt/β-catenin activation.
研究领域:
干细胞生物学
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骨生物学
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分子生物学
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再生医学
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产品关联
DOI
PubMed
高影响因子
Autophagy
·
2026
·
IF 18.6
Zhangjie Wang,Chaoyi Li,Ruiqiu Zhu,Minghao Duan,Zhenyu Kuang,Xin Li,Weifang Cui,Chaojun Duan,Chunfang Zhang
Lactylation is an emerging post-translational modification that is well established for its involvement in epigenetic regulation. However, its functional significance and regulatory mechanisms in non-small cell lung cancer (NSCLC) remain poorly understood. In this study, integrated proteomic and lactylomic analyses of clinical NSCLC specimens and matched adjacent normal tissues showed that USP7 (ubiquitin specific peptidase 7) was upregulated and USP7 K1084 lactylation was increased in NSCLC. Knockout of USP7 or inhibition of USP7 lactylation impaired cellular mitophagy, resulting in mitochondrial damage and attenuated NSCLC tumorigenicity. We identified CREBBP/CBP (CREB binding lysine acetyltransferase) as the key lactyltransferase responsible for USP7 K1084 lactylation. USP7 lactylation induced its localization to mitochondria and increased its interaction with PINK1 (PTEN induced kinase 1), promoting PINK1 deubiquitination and stabilization. Pharmacological inhibition of CREBBP reduced USP7 lactylation levels and promoted PINK1 ubiquitination and degradation, exerting antitumor effects in vitro and in vivo. Analysis of NSCLC clinical specimens showed that the protein levels of USP7, CREBBP, and PINK1 were positively correlated, and their high expression was associated with poor patient prognosis. Collectively, our findings establish the CREBBP-USP7-PINK1 axis as a promising therapeutic target for NSCLC treatment.Abbreviations: CHX: cycloheximide; CREBBP/CBP: CREB binding lysine acetyltransferase; DUB: deubiquitinating enzyme; IHC: immunohistochemistry; IP: immunoprecipitation; KO: knockout; LDHA: lactate dehydrogenase A; MS: mass spectrometry; NALA: L-sodium lactate; NSCLC: non-small cell lung cancer; OCR: oxygen consumption rate; PBS: phosphate-buffered saline; PINK1: PTEN induced kinase 1; PLA: proximity ligation assay; ROS: reactive oxygen species; shRNA: short hairpin RNA; TEM: transmission electron microscopy; TUBE: tandem ubiquitin binding entity; UPS: ubiquitin-proteasome system; USP7: ubiquitin specific peptidase 7; WT: wild type.
✓ 已验证
DOI
高影响因子
Cell Reports
·
2024
·
IF 7.5
Xiaohan Sun,Dafa Zhou,Yuanfei Sun,Yunhe Zhao,Yanran Deng,Xiaolin Pang,Qingxin Liu,Zizhang Zhou
The transcriptional coactivator Yorkie (Yki) regulates organ size by promoting cell proliferation. It is unclear how cells control Yki activity when exposed to harmful stimuli such as oxidative stress. In this study, we show that oxidative stress inhibits the binding of Yki to Scalloped (Sd) but promotes the interaction of Yki with another transcription factor, forkhead box O (Foxo), ultimately leading to a halt in cell proliferation. Mechanistically, Foxo normally exhibits a low binding affinity for Yki, allowing Yki to form a complex with Sd and activate proliferative genes. Under oxidative stress, Usp7 deubiquitinates Foxo to promote its interaction with Yki, thereby activating the expression of proliferation suppressors. Finally, we show that Yki is essential for Drosophila survival under oxidative stress. In summary, these findings suggest that oxidative stress reprograms Yki from a proliferation-promoting factor to a proliferation suppressor, forming a self-protective mechanism.
研究领域:
细胞生物学
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分子生物学
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发育生物学
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产品关联
DOI
PubMed
高影响因子
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY
·
2022
·Vol. 49
·期 1
·86-100
·
IF 1.254
Lingling C.,Hao W.,Fuqiang Y.,Chao G.,Honglin D.,Xiaojie S.,Yang Z.,Jiaxin Z.,Lihong S.,Hongmin L.,Qiurong Z.
Abstract Design and synthesis of two series of trifluoromethyl-containing polysubstituted pyrimidine derivatives in order to seek more effective anti-tumor drugs, and the antiproliferative activity of these compounds against four human tumor cell lines (PC-3, MGC-803, MCF-7, HGC-27) was evaluated by MTT method. Most of the compounds have moderate anti-proliferative activities, among which compounds ( XXId ) and ( XXIe ) have good selectivity for PC-3 cells. The IC 50 (concentration required to achieve 50% inhibition of the tumor cell proliferation) values of compounds ( XXId ) and ( XXIe ) for PC-3 cell were 4.42 ± 0.46 μmol L –1 and 4.85 ± 0.59 μmol L –1 , respectively, and their anti-proliferative activities were significantly better than that of the positive control 5-fluorouracil (6.39 ± 0.71 μmol L –1 ). Molecular docking results indicated that compounds ( XXId ) and ( XXIe ) were likely to be well integrated in the hydrophobic pocket of ubiquitin-specific protease 7 (USP7), which has the value of further research and development.
研究领域:
药物化学
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肿瘤学
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医学
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产品关联
DOI
高影响因子
BRITISH JOURNAL OF CANCER
·
2026
·
IF 7.8
Huang Yue,Xi Yiling,Nie Huizong,Wang Yan,Hu Jiahui,Gao Yating,Li Junhui,Ma Yaofang,Chu Jiahao,Tu Linglan,Huang Dongsheng,Cheng Liyan
Background LRRC41 is critical for the progression of multiple cancers, especially hepatocellular carcinoma (HCC), yet its oncogenic mechanism in HCC remains unclear. This study aimed to explore the molecular mechanism by which LRRC41 promotes HCC malignancy and investigate the therapeutic potential of inhibiting its mediated signalling pathway. Methods LRRC41 expression in HCC was analysed via the TCGA, HPA databases and experimental detection. Its biological functions were assessed by in vitro CCK-8, Transwell, colony formation assays and in vivo xenograft models. IP–MS, Co-IP and ubiquitination assays were used to elucidate the molecular mechanism, and the anti-tumour effect of the USP7 inhibitor P6620 was verified in vitro and in vivo. Results LRRC41 was abnormally overexpressed in HCC and correlated with poor prognosis, promoting HCC cell proliferation, invasion and tumorigenesis. USP7 stabilised LRRC41 via deubiquitination, and LRRC41 activated the NF-κB pathway by targeting HNRNPC in a K63-linked ubiquitination-dependent manner. P6620 inhibited the USP7/LRRC41 axis to suppress HCC malignancy, and its combination with lenvatinib enhanced in vivo anti-tumor efficacy. Conclusions LRRC41 overexpression drives HCC progression and poor prognosis; the novel USP7/LRRC41/HNRNPC/NF-κB axis is identified in HCC. USP7 inhibitor P6620 blocks this axis and may serve as a potential agent for HCC combination chemotherapy. The regulatory mechanism of the USP7/LRRC41/HNRNPC/NF-κB signalling axis in HCC. Created with Figdraw.
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DOI
PubMed
高影响因子
Frontiers in Pharmacology
·
2022
·
IF 5.988
Gu Yu-Hui,Ren Kai-Wen,Wang Yu,Wang Shi-Hao,Yu Xiao-Hong,Xu Li-Wen,Li Hui-Hua,Bi Hai-Lian
Hypertension is one of the common causes of pathological cardiac hypertrophy and a major risk for morbidity and mortality of cardiovascular diseases worldwide. Ubiquitin-Specific Protease 7 (USP7), the first identified deubiquitinating enzymes, participated in a variety of biological processes, such as cell proliferation, DNA damage response, tumourigenesis, and apoptosis. However, its role and mechanism in cardiac remodeling remain unclear. Here, our data indicated that USP7 expression was increased during Ang II-induced cardiac hypertrophy and remodeling in mice and humans with heart failure, while the administration of its inhibitor p22077 attenuated cardiac hypertrophy, cardiac fibrosis, inflammation, and oxidase stress. Mechanistically, the administration of p22077 inhibited the multiple signaling pathways, including AKT/ERK, TGF-β/SMAD2/Collagen I/Collagen III, NF-κB/NLRP3, and NAPDH oxidases (NOX2 and NOX4). Taken together, these findings demonstrate that USP7 may be a new therapeutic target for hypertrophic remodeling and HF.
研究领域:
心脏病学
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药理学
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分子生物学
✓ 已验证
DOI
PubMed
高影响因子
MOLECULAR THERAPY
·
2026
·
IF 11.4
Ye Chen,Jiaxin Bei,Xinkun Huang,Yujun Cai,Han Xu,Kangshun Zhu,Yihu Li,Chengyu Jia,Jiabai Huang,Zebing Song,Yongjian Guo,Zhe-Sheng Chen,Weiguo Cai,Bixiang Zhan,Mingyue Cai,Yin Zhang,Wensou Huang
Insufficient microwave ablation (iMWA) of hepatocellular carcinoma (HCC) often leads to rapid and aggressive recurrence, and effective intervention remains a formidable challenge. Here, we show that the DNA methylation machinery is aberrantly activated in residual HCC following iMWA. The heat-responsive stress protein HSP90α collaborates with the deubiquitinase USP7 to stabilize and upregulate DNA methyltransferase DNMT1, thereby transducing the sublethal heat-stress input from iMWA into an epigenetic output and elevating DNA methylation levels. Sublethal heat-stress-induced DNMT1 upregulation significantly promotes the growth and invasive potential of post-iMWA residual HCC cells, a process that involves disruption of propionate metabolism. Mechanistically, the propionate-metabolizing enzyme ACSS3 in residual HCC is found to be silenced by DNMT1-mediated promoter hypermethylation, impairing propionate-to-propionyl-coenzyme A (CoA) conversion. Reduction in propionyl-CoA production enhances the activity of the fatty acid β-oxidation (FAO) rate-limiting enzyme carnitine palmitoyltransferase 1 (CPT1), ultimately stimulating FAO-dependent ATP regeneration. Critically, we developed a liposomal CRISPR-dCas9-based locus-specific demethylation system for precise epigenetic reactivation of ACSS3 and demonstrated its ability to effectively prevent post-iMWA HCC recurrence and metastasis, with more pronounced efficacy when combined with propionate supplementation. Our findings identify a novel DNMT1-driven “epigenetic-metabolic cascade” as a central mechanism promoting HCC recurrence following iMWA, emphasizing the therapeutic potential of ACSS3 -targeted methylation editing.
✓ 已验证
DOI
PubMed
高影响因子
CANCER GENE THERAPY
·
2026
·
IF 6.4
Shi Ruxue,Gu Kai,Li Haichuan,Sun Qingwei,Wu Xinyu,Huang Shuhong,Hu Qingxia
Castration-resistant prostate cancer (CRPC) is one of the most prevalent cancers in men. The new generation androgen receptor (AR) inhibitor enzalutamide can improve the therapeutic effectiveness of patients with CRPC. However, these patients eventually develop acquired enzalutamide resistance (ENZR), and the mechanisms underlying resistance are not well understood. Wilms’ tumor 1-associating protein (WTAP) plays an important role in m 6 A modification and has been reported as an oncogene in various cancers. Here, we utilized a tissue microarray and collected tissues from prostate cancer (PCa) patients to detect WTAP expression, and found that WTAP is upregulated in PCa. Meanwhile, WTAP overexpression promotes cell proliferation and accelerates tumor growth through colony formation assays and the establishment of a subcutaneous xenograft model in vivo. These findings establish the tumor promoter role of WTAP in prostatic tumorigenesis. Furthermore, we verified that WTAP is a novel responsive gene of AR via promoter activity and chromatin immunoprecipitation (ChIP) assays. Importantly, we uncovered that WTAP is upregulated in ENZR cells, and WTAP knockdown inhibited the proliferation of ENZR cells. Mechanistically, ubiquitin-specific protease (USP7) enhanced the stability of WTAP by the ubiquitin-proteasome pathway in ENZR cells, thereby WTAP increases promote AKT signaling through an m 6 A-mediated way, and an AKT inhibitor can abolish the pro-resistance phenotype mediated by WTAP. Together, these findings suggest that WTAP plays a key role in ENZR development of PCa cells, and WTAP may be a potential treatment target for ENZR tumors. The alternative text for this image may have been generated using AI. In this manuscript, we utilized a tissue microarray and collected tissues from prostate cancer (PCa) patients to detect WTAP expression, and found that WTAP is upregulated in PCa. Meanwhile, WTAP overexpression promotes cell proliferation and accelerates tumor growth through colony formation assays and the establishment of a subcutaneous xenograft model in vivo. These findings establish the tumor promoter role of WTAP in prostatic tumorigenesis. Furthermore, we verified that WTAP is a novel responsive gene of AR via promoter activity and chromatin immunoprecipitation (ChIP) assays. Importantly, we uncovered that WTAP is upregulated in ENZR cells, and WTAP knockdown inhibited the proliferation of ENZR cells. Mechanistically, ubiquitin-specific protease (USP7) enhanced the stability of WTAP by the ubiquitin-proteasome pathway in ENZR cells, thereby WTAP increases promote AKT signaling through an m 6 A-mediated way, and an AKT inhibitor can abolish the pro-resistance phenotype mediated by WTAP. Together, these findings suggest that WTAP plays a key role in ENZR development of PCa cells, and WTAP may be a potential treatment target for ENZR tumors.
✓ 已验证
DOI
PubMed
高影响因子
CANCER RESEARCH
·
2025
·
IF 16.6
Zhang Jie,Chen Weiwu,Zhang Chen,He Qian,Wang Xudong,Han Jiayu,Gao Peng,Wang Kunyu,Xie Haoze,Gao Feng,Guo Yining,Guo Wenhao,Jiang Haofei,Le Jing,Zang Ruichen,Mo Sisi,Fan Haobo,Zhu Xiaoxiao,Jiang Xinrong,Gao Fengbin,Yu Yanlan,Ding Guoqing,Chen Yicheng
PD-1 restrains effective killing of cancer cells by the immune system and is predominantly located on the surface of T cells or other immune cells. However, cancer cells also express PD-1 to varying degrees, which is commonly associated with a poor prognosis. Here, we investigated the regulation and function of PD-1 expression in prostate cancer (PCa) and revealed the impact on the tumor microenvironment. PD-1 expression in cancer cells positively correlated with Gleason grade and metastasis but negatively correlated with CD8+ T cell infiltration in PCa patients. PCa cells secreted PD-1 in exosomes that enhanced the activity of myeloid-derived suppressor cells (MDSCs) by activating JAK/STAT3 signaling. The activated MDSCs in turn reduced the infiltration of CD8+ T cells within the tumor, promoting tumor immune evasion. The ubiquitin-specific protease USP7 induced deubiquitination and elevated the abundance of PD-1 in PCa, and USP7 inhibition sensitized PCa tumors to anti-PD-1 antibody treatment. Given the modest efficacy of current immunotherapeutic approaches for PCa, strategies to inhibit the secretion of PD-1-bearing exosomes or USP7 function may emerge as promising immunostimulatory interventions for treating PCa.
研究领域:
癌症生物学
·
免疫学
·
分子生物学
·
细胞信号传导
·
泌尿学
·
肿瘤学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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