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经历了明显的代谢改变。最近的 研究表明，功能性线粒体对于肿瘤细胞发育至关重要。但是，我们很大程度 不知道线粒体因子如何导致转移性进展。识别新的线粒体因子 对于癌症转移至关重要，我们使用自定义的RNAi库进行了公正的基于RNAi的屏幕 靶向120个线粒体酶。使用各种类型的癌细胞筛选氨基磷酸酯 - 糖charopine途径中的半甲醛合酶（AASS）是入侵的常见关键因素。体外 使用AASS变体的体内研究表明，AASS的活性是癌细胞浸润所必需的 和转移。针对糖含量途径中的每种酶，均显示2-氨基磷酸盐（2-AAA）， 代谢中间体，是细胞浸润的关键因素。代谢，基因组和转录组学方法 被制作以获得对癌症转移中AASS的机械洞察力。通过一系列代谢测定， 我们发现AASS通过在肺部和头部/颈部癌症中管理氧化还原稳态来促进入侵。 基因组分析和GSEA表明，AASS损失导致过氧化物酶体基因的降低和氧化还原 蛋白质TXNDC5。此外，XBP1的拼接形式是TXNDC5的激活转录因子 缺乏AASS的细胞，表明AASS，SXBP1和TXNDC5之间存在潜在的联系。另外，通过 激酶抑制剂分析和全面的转录因子分析，我们确定了上游调节剂 aass。特别是，我们发现HGF/C-MET作为激酶和AASS的激活剂，而SATB1则是一种潜力 AASS的转录因子。此外，我们的初步数据表明AASS上调 与癌症患者的肿瘤组织转移性进展相关，表明AASS是一种有希望的 预后标记和治疗转移性癌症的治疗靶标。因此，我们筛选并确定了 美国FDA批准了复合单宁酸作为有效的AASS抑制剂和抗碎裂剂。 我们的中心假设是AASS及其代谢中间体2-AAA提供转移信号 通过它们的潜在调节剂和癌细胞的效应子。因此，AASS信号代表一个有前途的 人类癌症中的抗近时间靶标。我们将使用肺癌和头颈癌作为研究 验证AASS信号作为常见转移驱动器的平台。提出了三个具体目标：（1）到 破译AASS-XBP1激活的分子机制，该机制管理氧化还原信号和 赋予人类癌症中的转移性潜力； （2）确定HGF/C-如何激活和诱导AASS 大都会和satb1介导转移； （3）验证AASS作为治疗转移性的治疗靶标 使用新型AASS抑制剂单宁酸的癌症。