Ref-1 redox activity alters cancer cell metabolism in pancreatic cancer: exploiting this novel finding as a potential target.
Ref-1 redox activity alters cancer cell metabolism in pancreatic cancer: exploiting this novel finding as a potential target.
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Ref-1氧化还原活性改变胰腺癌中的癌细胞代谢：利用这一新发现作为潜在靶点
DOI：
10.1186/s13046-021-02046-x
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发表时间：
2021-08-10
期刊：
影响因子：
--
通讯作者：
Fishel ML
中科院分区：
文献类型：
--
作者：
Gampala S;Shah F;Lu X;Moon HR;Babb O;Umesh Ganesh N;Sandusky G;Hulsey E;Armstrong L;Mosely AL;Han B;Ivan M;Yeh JJ;Kelley MR;Zhang C;Fishel ML
胰腺癌是一种复杂的疾病，具有促结缔组织增生的间质、极度缺氧和对治疗的固有抵抗。了解这种侵袭性癌症的信号传导和适应性反应是提高治疗效果的关键。氧化还原因子-1（Ref-1）是一种氧化还原信号蛋白，可调节包括HIF-1 α、STAT3和NF κ B在内的多种转录因子（TF）从氧化态到还原态的转化，从而增强其与DNA的结合。在我们先前发表的工作中，在常氧条件下敲低Ref-1导致包括EIF2、蛋白激酶A和mTOR在内的通路上的基因表达模式改变。本研究采用单细胞RNA测序技术（scRNA-seq）和蛋白质组学技术，探讨缺氧条件下Ref-1对代谢途径的影响。使用左截断混合高斯模型分析比较表达少于20%的Ref-1蛋白的胰腺癌细胞的scRNA-seq，并使用蛋白质组学和qRT-PCR验证。利用线粒体功能测定、qRT-PCR、western blotting和NADP测定证实了所鉴定的Ref-1在线粒体功能中的作用。此外，使用体外3D共培养和体内异种移植研究测定Ref-1氧化还原功能抑制对胰腺癌代谢的作用。从scRNA-seq数据中，在Ref-1敲低与加扰对照中鉴定了中枢代谢、细胞周期、细胞凋亡、免疫应答以及一系列信号传导途径和转录调控因子下游基因中的独特转录变异。在Ref-1氧化还原抑制后，Ref-1敲低下调的线粒体DEG亚群显著减少，并且在体外与Devimistat组合时更显著。线粒体功能测定表明，Ref-1敲低和Ref-1氧化还原信号传导抑制降低了TCA循环底物的利用，并减缓了胰腺癌共培养球体的生长。在Ref-1敲减细胞中，观察到NADP+消耗反应的较高通量率，表明这些细胞中NADP+的可用性较低，氧化应激水平较高。体内异种移植物研究表明，与Devimistat相似的Ref-1氧化还原抑制剂可有效减少肿瘤。Ref-1氧化还原信号传导抑制通过引起TCA循环功能障碍而最终改变癌细胞代谢，同时还在体外以及体内减少胰腺肿瘤生长。在线版本包含补充材料，可通过10.1186/s13046 - 021 - 02046-x获得。
Pancreatic cancer is a complex disease with a desmoplastic stroma, extreme hypoxia, and inherent resistance to therapy. Understanding the signaling and adaptive response of such an aggressive cancer is key to making advances in therapeutic efficacy. Redox factor-1 (Ref-1), a redox signaling protein, regulates the conversion of several transcription factors (TFs), including HIF-1α, STAT3 and NFκB from an oxidized to reduced state leading to enhancement of their DNA binding. In our previously published work, knockdown of Ref-1 under normoxia resulted in altered gene expression patterns on pathways including EIF2, protein kinase A, and mTOR. In this study, single cell RNA sequencing (scRNA-seq) and proteomics were used to explore the effects of Ref-1 on metabolic pathways under hypoxia. scRNA-seq comparing pancreatic cancer cells expressing less than 20% of the Ref-1 protein was analyzed using left truncated mixture Gaussian model and validated using proteomics and qRT-PCR. The identified Ref-1’s role in mitochondrial function was confirmed using mitochondrial function assays, qRT-PCR, western blotting and NADP assay. Further, the effect of Ref-1 redox function inhibition against pancreatic cancer metabolism was assayed using 3D co-culture in vitro and xenograft studies in vivo. Distinct transcriptional variation in central metabolism, cell cycle, apoptosis, immune response, and genes downstream of a series of signaling pathways and transcriptional regulatory factors were identified in Ref-1 knockdown vs Scrambled control from the scRNA-seq data. Mitochondrial DEG subsets downregulated with Ref-1 knockdown were significantly reduced following Ref-1 redox inhibition and more dramatically in combination with Devimistat in vitro. Mitochondrial function assays demonstrated that Ref-1 knockdown and Ref-1 redox signaling inhibition decreased utilization of TCA cycle substrates and slowed the growth of pancreatic cancer co-culture spheroids. In Ref-1 knockdown cells, a higher flux rate of NADP + consuming reactions was observed suggesting the less availability of NADP + and a higher level of oxidative stress in these cells. In vivo xenograft studies demonstrated that tumor reduction was potent with Ref-1 redox inhibitor similar to Devimistat. Ref-1 redox signaling inhibition conclusively alters cancer cell metabolism by causing TCA cycle dysfunction while also reducing the pancreatic tumor growth in vitro as well as in vivo. The online version contains supplementary material available at 10.1186/s13046-021-02046-x.
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影响因子：
5.7
作者：
Arpin CC;Mac S;Jiang Y;Cheng H;Grimard M;Page BD;Kamocka MM;Haftchenary S;Su H;Ball DP;Rosa DA;Lai PS;Gómez-Biagi RF;Ali AM;Rana R;Hanenberg H;Kerman K;McElyea KC;Sandusky GE;Gunning PT;Fishel ML
通讯作者：
Fishel ML
影响因子：
11.2
作者：
Chang, Qing;Jurisica, Igor;Hedley, David W.
通讯作者：
Hedley, David W.
影响因子：
2.8
作者：
Illuzzi JL;McNeill DR;Bastian P;Brenerman B;Wersto R;Russell HR;Bunz F;McKinnon PJ;Becker KG;Wilson DM 3rd
通讯作者：
Wilson DM 3rd
多功能APE1 DNA Repair-Redox信号蛋白是人类疾病中的药物靶标。
影响因子：
7.4
作者：
Caston RA;Gampala S;Armstrong L;Messmann RA;Fishel ML;Kelley MR
通讯作者：
Kelley MR
影响因子：
6.6
作者：
Chiu, Hsin Yao;Tay, Emmy Xue Yun;Taneja, Reshma
通讯作者：
Taneja, Reshma