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: 10551995
• 负责人: Sumin Kang
• 金额: $ 41.14万
• 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/10551995
• 关键词: Attenuated; Binding Proteins; Biological Assay; Cancer Patient; Cancer cell line; Cause of Death; Cells; Clinical; 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; Invaded; Libraries; Link; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of ovary; Mediating; Metabolic; 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 Promotion; Tumor Tissue; Up-Regulation; Variant; XBP1 gene; Xenograft procedure; activating transcription factor 1; 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会经历显著的代谢变化。近期 研究表明，功能性线粒体对肿瘤细胞的发育是必不可少的。然而，我们基本上是这样做的 不知道线粒体因素如何促进转移进展。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的无偏见筛查 以120个线粒体酶为靶点。使用各种类型的癌细胞筛选出令人陶醉的氨基己二酸酯- 糖胺途径中的半醛合成酶(AASS)是肿瘤侵袭的共同关键因素。离体 用AASS变异体进行的体内研究表明，AASS的活性是癌细胞侵袭所必需的 和转移。以糖胺途径中的每个酶为靶标显示2-氨基己二酸酯(2-AAA)， 一种代谢中间体，是细胞入侵的关键因素。代谢、基因组和转录组学方法 是为了从机制上深入了解AASS在癌症转移中的作用。通过一系列新陈代谢测试， 我们发现，AASS通过管理肺癌和头颈癌的氧化还原动态平衡来促进侵袭。 基因组图谱和GSEA分析表明，AASS缺失导致过氧化体基因减少和氧化还原 蛋白TXNDC5。此外，TXNDC5的激活转录因子XBP1的剪接形式在 缺乏AASS的细胞，提示AASS、sXBP1和TXNDC5之间存在潜在的联系。此外，通过一个 激酶抑制因子谱和综合转录因子谱，我们确定了 阿斯。特别是，我们发现hgf/c-met作为ass的一种激酶和激活剂，而sab1作为一种潜在的。 AASS转录因子。此外，我们的初步数据显示，AASS正向上调 与癌症患者肿瘤组织的转移进展相关，提示AASS是一种有前途的 预后标记物和治疗转移性癌症的靶点。因此，我们筛选并鉴定了一种 美国FDA批准复合单宁酸作为一种有效的AASS抑制剂和抗转移药物。 我们的中心假设是AASS及其代谢中间产物2-AAA提供转移信号 通过它们在癌细胞中的潜在调节器和效应器。因此，AASS信令代表着一种很有前途的 人类癌症中的抗转移靶点。我们将以肺癌和头颈癌为研究对象 验证AASS信号作为常见转移驱动因素的平台。提出了三个具体目标：(1) 破译AASS-XBP1激活的分子机制，它管理氧化还原信号和 赋予人类癌症转移潜能；(2)确定HGF/c-AAS是如何被激活和诱导的。 MET和SATB1介导转移；(3)验证AASS作为治疗转移性肿瘤的靶点 使用新型AASS抑制剂单宁酸治疗癌症。