Decoding and targeting saccharopine pathway in cancer metastasis

解码和靶向癌症转移中的糖碱途径

• 批准号: 10344916
• 负责人: Sumin Kang
• 金额: $ 41.98万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10344916
• 关键词: Attenuated; Binding Proteins; Biological Assay; Cancer Patient; Cancer cell line; Cause of Death; Cells; Clinical; Custom; Data; Development; Disseminated Malignant Neoplasm; Down-Regulation; Enzymes; Equilibrium; FDA approved; Gene set enrichment analysis; Genes; Genomics; Glean; HGF gene; Head Cancer; Head and Neck Cancer; Homeostasis; Human; In Vitro; Libraries; Link; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of ovary; Mediating; Metabolic; Metastatic to; Mitochondria; Molecular; Neck Cancer; Neoplasm Metastasis; Oxidation-Reduction; Pathway interactions; Patients; Phosphotransferases; Play; Prognostic Marker; Proteins; RNA Interference; RNA Splicing; RNA interference screen; Research; Series; Signal Pathway; Signal Transduction; Solid Neoplasm; TXN gene; Tannic Acid; Treatment Efficacy; Tumor Tissue; Up-Regulation; Variant; XBP1 gene; Xenograft procedure; activating transcription factor; cancer cell; cancer therapy; cancer type; cohort; in vivo; inhibitor; insight; kinase inhibitor; malignant breast neoplasm; neoplastic cell; novel; patient derived xenograft model; saccharopine; small hairpin RNA; small molecule; therapeutic target; transcription factor; transcriptome sequencing; transcriptomics; tumor; upstream kinase

Growing evidence suggests that cancer cells undergo significant metabolic alterations during metastasis. Recent studies show that functional mitochondria are essential for the tumor cell development. However, we largely do not know how mitochondrial factors contribute to metastatic progression. To identify a novel mitochondrial factor critical for cancer metastasis, we performed an unbiased RNAi-based screen using a customized RNAi library targeting 120 mitochondrial enzymes. Screen using various types of cancer cells reveled that aminoadipate- semialdehyde synthase (AASS) in the saccharopine pathway is a common critical factor for the invasion. In vitro and in vivo study using AASS variants demonstrated that the activity of AASS is required for cancer cell invasion and metastasis. Targeting each of the enzymes in the saccharopine pathway revealed 2-aminoadipate (2-AAA), a metabolic intermediate, as a key factor for cell invasion. Metabolic, genomic, and transcriptomics approaches were made to gain mechanistic insight into AASS in cancer metastasis. Through a series of metabolic assays, we found that AASS promotes invasion by managing redox homeostasis in lung and head/neck cancers. Genomic profiling and GSEA showed that AASS loss results in decrease of peroxisomal genes and a redox protein TXNDC5. Moreover, spliced form of XBP1, an activated transcription factor of TXNDC5, was reduced in cells that lack AASS, suggesting a potential link among AASS, sXBP1, and TXNDC5. In addition, through a kinase inhibitor profiling and comprehensive transcription factor profiling, we identified upstream regulators of AASS. In particular, we found HGF/c-MET as a kinase and an activator of AASS, and SATB1 as a potential transcription factor of AASS. Furthermore, our preliminary data showed that AASS upregulation positively correlates with metastatic progression in tumor tissues from cancer patients, suggesting AASS as a promising prognostic marker and a therapeutic target to treat metastatic cancers. Therefore, we screened and identified a US FDA approved compound tannic acid as a potent AASS inhibitor and an anti-metastasis agent. Our central hypothesis is that AASS and its metabolic intermediate 2-AAA provide metastastic signals through their potential regulators and effectors in cancer cells. Thus, AASS signaling represents a promising anti-metastasis target in human cancers. We will use lung cancer and head and neck cancer as a research platform to validate AASS signaling as a common metastasis driver. Three specific aims are proposed: (1) To decipher the molecular mechanism underlying AASS-XBP1 activation, which manages redox signaling and confers metastatic potential in human cancer; (2) To determine how AASS is activated and induced by HGF/c- MET and SATB1 to mediate metastasis; (3) To validate AASS as a therapeutic target in treatment of metastatic cancers using the novel AASS inhibitor tannic acid.

越来越多的证据表明，癌细胞在转移过程中发生了显著的代谢改变。最近 研究表明，功能性线粒体对于肿瘤细胞的发展是必不可少的。然而，我们在很大程度上 不知道线粒体因子如何促进转移进展。鉴定一种新的线粒体因子 对于癌症转移至关重要，我们使用定制的RNAi文库进行了基于RNAi的无偏筛选， 靶向120种线粒体酶使用各种类型的癌细胞进行的筛选显示，氨基己二酸- 半醛合酶（AASS）在糖嘌呤途径是一个共同的关键因素的入侵。体外 使用AASS变体的体内研究表明，AASS活性是癌细胞侵袭所必需的 和转移。靶向糖精途径中的每种酶显示2-氨基己二酸（2-AAA）， 一种代谢中间体，是细胞侵袭的关键因素。代谢、基因组和转录组学方法 目的：探讨AASS在肿瘤转移中的作用机制。通过一系列的代谢分析， 我们发现AASS通过控制肺癌和头颈癌中的氧化还原稳态来促进侵袭。 基因组分析和GSEA表明，AASS缺失导致过氧化物酶体基因的减少和氧化还原酶的减少。 TXNDC 5蛋白。此外，剪接形式的XBP 1，一种激活的转录因子TXNDC 5，减少， 缺乏AASS的细胞，这表明AASS，sXBP 1和TXNDC 5之间存在潜在联系。此外，通过A 激酶抑制剂分析和综合转录因子分析，我们确定了上游调控因子， AASS。特别是，我们发现HGF/c-MET是AASS的激酶和激活剂，而SATB 1是AASS的潜在激活剂。 AASS转录因子。此外，我们的初步数据表明，AASS上调正 与癌症患者肿瘤组织中的转移进展相关，表明AASS是一种有前途的 预后标志物和治疗转移性癌症的治疗靶点。因此，我们筛选并鉴定了 美国FDA批准复方鞣酸作为一种有效的AASS抑制剂和抗转移剂。 我们的中心假设是，AASS和它的代谢中间体2-AAA提供了代谢信号 通过它们在癌细胞中的潜在调节器和效应器。因此，AASS信号转导代表了一种有前途的 抗转移靶点。我们将以肺癌和头颈癌为研究对象， 平台来验证AASS信号传导作为常见的转移驱动因素。提出了三个具体目标：（1） 破译AASS-XBP 1激活的分子机制，该机制管理氧化还原信号传导， （2）为了确定AASS如何被HGF/c激活和诱导， MET和SATB 1介导肿瘤转移;（3）验证AASS作为肿瘤转移治疗靶点的可行性 使用新型AASS抑制剂单宁酸治疗癌症。