Deubiquitinase Activity Profiling Identifies UCHL1 as a Candidate Oncoprotein That Promotes TGFβ-Induced Breast Cancer Metastasis.
Deubiquitinase Activity Profiling Identifies UCHL1 as a Candidate Oncoprotein That Promotes TGFβ-Induced Breast Cancer Metastasis.
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DOI：
10.1158/1078-0432.ccr-19-1373
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发表时间：
2020-03-15
期刊：
影响因子：
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通讯作者：
Ten Dijke P
中科院分区：
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作者：
Liu S;González-Prieto R;Zhang M;Geurink PP;Kooij R;Iyengar PV;van Dinther M;Bos E;Zhang X;Le Dévédec SE;van de Water B;Koning RI;Zhu HJ;Mesker WE;Vertegaal ACO;Ovaa H;Zhang L;Martens JWM;Ten Dijke P
针对特异性分子靶点的治疗对于三阴性乳腺癌（TNBC）患者仍然未得到满足。去泛素化酶（DUBs）是新兴的药物靶点。TNBC中高活性DUB的鉴定可能导致新的疗法。使用DUB活性探针，我们分析了52个乳腺癌细胞系和52个患者肿瘤组织中的总体DUB活性。为了在体内验证我们的发现，我们采用了斑马鱼和小鼠乳腺癌异种移植模型。利用体内和体外生物化学方法阐明了细胞和分子机制。合成了一种特异性抑制剂，并在一系列试验中评估了其生化和生物学功能。最后，我们使用患者血清样本来研究临床相关性。两种DUB活性分析方法将UCHL1鉴定为在TNBC细胞系和侵袭性肿瘤中具有高度活性。在功能上，UCHL1促进了斑马鱼和小鼠乳腺癌异种移植模型的转移。在机制上，UCHL1通过保护TGFβ I型受体和SMAD2免于泛素化而促进TGFβ信号传导诱导的转移。我们发现这些反应被特异性UCHL1抑制剂6RK73有效抑制。此外，UCHL1水平在TNBC患者血清中显著增加，并且在血清外泌体以及TNBC细胞条件培养基中高度富集。UCHL1富集的外泌体刺激乳腺癌迁移和外渗，表明UCHL1可能以旁分泌方式促进肿瘤进展。我们的DUB活性分析将UCHL1鉴定为促进TGFβ诱导的乳腺癌转移的候选癌蛋白，并可能为TNBC治疗提供潜在靶点。
Therapies directed to specific molecular targets are still unmet for triple-negative breast cancer (TNBC) patients. Deubiquitinases (DUBs) are emerging drug targets. The identification of a highly active DUBs in TNBC may lead to novel therapies. Using DUB activity probes, we profiled global DUB activities in 52 breast cancer cell lines and 52 patients’ tumor tissues. To validate our findings in vivo, we employed both zebrafish and murine breast cancer xenograft models. Cellular and molecular mechanisms were elucidated using in vivo and in vitro biochemical methods. A specific inhibitor was synthesised and its biochemical and biological functions were assessed in a range of assays. Finally, we used patient sera samples to investigate clinical correlations. Two DUB activity profiling approaches identified UCHL1 as being highly active in TNBC cell lines and aggressive tumors. Functionally, UCHL1 promoted metastasis in zebrafish and murine breast cancer xenograft models. Mechanistically, UCHL1 facilitates TGFβ signaling-induced metastasis by protecting TGFβ type I receptor and SMAD2 from ubiquitination. We found that these responses are potently suppressed by the specific UCHL1 inhibitor, 6RK73. Furthermore, UCHL1 levels were significantly increased in TNBC patient sera, and highly enriched in sera exosomes as well as TNBC cell conditioned media. UCHL1 enriched exosomes stimulated breast cancer migration and extravasation, suggesting that UCHL1 may act in a paracrine manner to promote tumor progression. Our DUB activity profiling identified UCHL1 as a candidate oncoprotein that promotes TGFβ-induced breast cancer metastasis and may provide a potential target for TNBC treatment.