A positive feedback loop regulation between NOTCH1 and USP11 in T-cell leukemia.
A positive feedback loop regulation between NOTCH1 and USP11 in T-cell leukemia.
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DOI：
10.1038/s41375-023-02096-4
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发表时间：
2024-01
期刊：
影响因子：
11.4
通讯作者：
Ntziachristos, Panagiotis
中科院分区：
文献类型：
--
作者：
Fijalkowski, Igor;Wang, Jin;Jin, Qi;Van Laere, Jolien;Serafin, Valentina;Crispino, John D.;Ntziachristos, Panagiotis
NOTCH 1是一种高度保守的信号通路中的跨膜受体，其控制各种生物体和组织中的细胞命运决定。相反，其失调与发育障碍转化相关[1]。NOTCH 1由细胞外和细胞内结构域组成。配体与NOTCH 1胞外区结合后，分别由ADAM家族金属蛋白酶和γ-分泌酶介导两个连续的切割步骤，产生有活性的NOTCH 1胞内区，并与其他转录辅激活因子协同作用，激活下游基因表达。值得注意的是，超过60%的T细胞淋巴母细胞白血病（T-ALL）病例中报告了NOTCH 1突变的功能获得。因此，控制NOTCH 1信号传导的分子机制的解剖可能提供新的治疗窗口。600种泛素E3连接酶调节泛素化，并可被约100种去泛素化（DUB）酶逆转[2]。先前的研究强调了由泛素特异性蛋白酶（USP）介导的去泛素化在稳定T-ALL或血管生成发芽的背景下的NOTCH 1中的关键作用[3，4]。我们最近证明了USP 7在这种疾病中去泛素化和稳定NOTCH 1蛋白水平的作用[4]以及去泛素化在治疗抗性中的作用[5]。来自儿科癌症基因组计划（PeCan）的生存数据以及52个USP家族成员的表达水平表明，USP 11水平与不良预后相关[5]。在高风险疾病病例的诊断样本中，USP 11的蛋白水平显著升高，这表明它可能与对治疗的耐药性有关[5]。为了更好地证明USP 11在调节NOTCH 1信号传导中的功能，我们利用生物化学和分子测定结合癌症数据集的生物信息学分析来揭示T-ALL中NOTCH 1和USP 11之间的新的正反馈回路。在T-ALL中，NOTCH 1表达经常通过突变、转录和翻译后调节激活[1]。USP 7和NOTCH 1存在一个前馈循环，NOTCH 1转录启动USP 7基因座[4]。我们首先分析了儿科癌症基因组计划（PeCan）数据库，以确定52个USP和NOTCH 1在mRNA水平上的相关性（图1A）。在这52个USP中，USP 11被鉴定为与NOTCH 1转录物正相关的转录物（图1A、B）。对癌细胞系百科全书（CCLE）数据库的进一步分析显示，发现与来自其他癌症类型的细胞系相比，NOTCH 1和USP 11在T-ALL细胞系中共同高度表达（图1C），并且它们的水平在T-ALL组中呈现正相关（图1D）。为了进一步证实NOTCH 1和USP 11表达之间的正相关性，我们在T-ALL患者样本中进行了反相蛋白阵列（RPPA）分析。我们的分析显示，N 0 TCH 1蛋白水平与USP 11蛋白水平正相关（图1 E）。总之，这些结果表明T-ALL中USP 11和NOTCH 1表达之间呈正相关。接下来，我们质疑NOTCH 1是否转录调控USP 11。首先，我们分析了T-ALL细胞中USP 11基因座上NOTCH 1、BRD 4和H3 K27 Ac的染色质免疫沉淀结合下一代测序（ChIP-seq）数据。值得注意的是，在与H3 K27 Ac信号相关的USP 11启动子上鉴定到NOTCH 1峰，表明NOTCH 1对USP 11的潜在转录调节（图1F）。此外，γ-分泌酶抑制剂（γSI）介导的。
NOTCH1 is a transmembrane receptor in a highly conserved signaling pathway that controls cell fate decisions in various organisms and tissues. In contrast, its dysregulation is associated with developmental disorders transformation [1]. NOTCH1 is composed of extracellular and intracellular domains. Ligand binding of NOTCH1 extracellular domain leads to two consecutive cleavage steps mediated by ADAM-family metalloproteases and γ-secretase respectively, to generate active NOTCH1 intracellular domain, which orchestrated with other transcriptional coactivators, activates downstream gene expression. Notably, gain of function NOTCH1 mutations have been reported in more than 60% of T cell lymphoblastic leukemia (T-ALL) cases. Therefore, the dissection of the molecular mechanisms controlling NOTCH1 signaling might provide novel therapeutic windows.> 600 ubiquitin E3 ligases regulate ubiquitination and can be reversed by about 100 deubiquitinating (DUB) enzymes [2]. Previous studies highlighted the critical role of deubiquitination mediated by ubiquitin-specific proteases (USP) in stabilizing NOTCH1 both in the context of T-ALL or angiogenic sprouting [3, 4]. We recently demonstrated the role of USP7 in deubiquitinating and stabilizing NOTCH1 protein levels in this disease [4] and the role of deubiquitination in therapy resistance [5]. Survival data from the pediatric cancer genome project (PeCan) coupled with the expression levels of 52 USP family members showed that USP11 levels are associated with poor prognosis [5]. Protein levels of USP11 are significantly elevated in diagnostic samples of highrisk disease cases, suggesting it might be associated with resistance to therapy [5]. To better demonstrate the function of USP11 in regulating NOTCH1 signaling, we utilized biochemistry and molecular assays coupled with bioinformatic analysis of cancer datasets to reveal a novel positive feedback loop between NOTCH1 and USP11 in T-ALL. NOTCH1 expression is frequently activated via mutation, transcriptional, and posttranslational regulation in T-ALL [1]. USP7 and NOTCH1 present with a feed-forward loop, with NOTCH1 transcriptionally starting the USP7 locus [4]. We first analyzed the pediatric cancer genome project (PeCan) database to determine the correlation between the 52 USPs and NOTCH1 at the mRNA level (Fig. 1 A). Among those 52 USPs, USP11 was identified as transcript showing positive correlation with NOTCH1 transcript (Fig. 1 A, B). Further analysis of the Cancer Cell Line Encyclopedia (CCLE) database showed that both NOTCH1 and USP11 were found to be collectively highly expressed in T-ALL cell lines compared with cell lines from other cancer types (Fig. 1 C), and their levels present with a positive correlation in the T-ALL group (Fig. 1 D). To further confirm the positive correlation between NOTCH1 and USP11 expression, we performed reverse-phase protein array (RPPA) analysis in T-ALL patient samples. Our analysis shows that NOTCH1 protein levels positively correlate with USP11 protein levels (Fig. 1 E). Together, these results demonstrate a positive correlation between USP11 and NOTCH1 expression in T-ALL. Next, we questioned whether NOTCH1 transcriptionally regulates USP11. First, we analyzed chromatin immunoprecipitation coupled to next-generation sequencing (ChIP-seq) data of NOTCH1, BRD4, and H3K27Ac on the USP11 locus in T-ALL cells. Of note, NOTCH1 peaks were identified on the USP11 promoter associated with the H3K27Ac signal suggesting the potential transcriptional regulation of USP11 by NOTCH1 (Fig. 1 F). Furthermore, γ-secretase inhibitor (γSI)-mediated …
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影响因子：
13.6
作者：
通讯作者：
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DOI：
10.1073/pnas.1315023111
发表时间：
2014-01-14
影响因子：
11.1
作者：
Wang, Hongfang;Zang, Chongzhi;Aster, Jon C.
通讯作者：
Aster, Jon C.
影响因子：
82.9
作者：
Kourtis N;Lazaris C;Hockemeyer K;Balandrán JC;Jimenez AR;Mullenders J;Gong Y;Trimarchi T;Bhatt K;Hu H;Shrestha L;Ambesi-Impiombato A;Kelliher M;Paietta E;Chiosis G;Guzman ML;Ferrando AA;Tsirigos A;Aifantis I
通讯作者：
Aifantis I
影响因子：
50.3
作者：
Ntziachristos P;Lim JS;Sage J;Aifantis I
通讯作者：
Aifantis I
影响因子：
56.9
作者：
Lim, R.;Sugino, T.;Potente, M.
通讯作者：
Potente, M.