Nicotinamide N-Methyltransferase (NNMT) as a master regulator of cancer stroma

烟酰胺 N-甲基转移酶 (NNMT) 作为癌症基质的主要调节因子

• 批准号: 9382387
• 负责人: Ernst Lengyel
• 金额: $ 43.2万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-15 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9382387
• 关键词: Abdominal Cavity; Address; Adhesions; Affect; Alternative Splicing; Biochemical; Biological Assay; Cancer Cell Growth; Cancer Patient; Cell Adhesion; Cell physiology; Cells; Chemicals; Data; Enzymes; Epigenetic Process; Epithelial; Epithelial ovarian cancer; Extracellular Matrix; Fibroblasts; Gene Expression; Genes; Genetic Transcription; Genomics; Goals; Growth; Growth Factor; Histones; Human; Immunocompetent; In Vitro; Label; Lead; Light; Maintenance; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Mesenchymal; Metabolic; Metabolic Pathway; Metabolism; Neoplasm Metastasis; Niacinamide; Nicotinamide N-Methyltransferase; Normal Cell; Oncogenic; Oxidative Phosphorylation; Patients; Pattern; Phenotype; Proteins; Proteomics; Role; S-Adenosylhomocysteine; Sampling; Serous; System; Translational Research; Tumor Burden; Tumor Promotion; United States National Institutes of Health; Work; Xenograft procedure; base; cancer cell; cell type; clinically relevant; cytokine; differential expression; experimental study; genome wide methylation; high throughput screening; histone methylation; in vivo; inhibitor/antagonist; knock-down; laser capture microdissection; metabolomics; methyl group; mouse model; neoplastic cell; novel; outcome forecast; overexpression; protein function; small molecule inhibitor; targeted treatment; three dimensional cell culture; three-dimensional modeling; transcriptome; tumor; tumor growth; tumor microenvironment; tumor progression; tumorigenesis

ABSTRACT Serous epithelial ovarian cancer (OvCa) typically presents with wide dissemination of cancer cells within the abdominal cavity and significant tumor burden. OvCa metastases have a high proportion of stroma, which consists primarily of cancer associated fibroblasts (CAFs), a mesenchymal cell type known to promote the invasion and metastasis of tumor cells. However, it is unclear how normal fibroblasts are reprogrammed into CAFs and how CAFs promote tumor growth. To directly address these open questions, we performed proteomic analyses of the primary and metastatic stroma in OvCa patient samples to identify proteins that were differentially expressed. We detected, specifically in the metastases, a conserved stromal signature associated with metastasis that included high stromal expression of Nicotinamide N-Methyltransferase (NNMT). NNMT catalyzes the transfer of a methyl group from S-adenosyl methionine (SAM) to nicotinamide. This depletes cellular SAM stores and leads to global hypomethylation of histones and expression of tumor-promoting genes. In preliminary experiments, we found that NNMT can reprogram normal fibroblasts into CAFs and that NNMT inhibition in CAFs blocks OvCa cancer cell adhesion, proliferation, and in vivo tumor growth. In our system, NNMT expression specifically regulates trimethylation of H3K4 and H3K27. Analysis of NNMT expression in OvCa patients revealed that strong stromal NNMT expression is significantly associated with a poor prognosis. Based on these data, the primary hypothesis underlying this application is that expression of NNMT in normal fibroblasts transforms them to CAFs through metabolically-mediated epigenetic alterations. The proposed experiments will systematically characterize the contribution of NNMT-driven epigenetic and metabolic changes to the transformation of normal fibroblasts to CAFs. In Aim I, we propose to investigate NNMT-driven epigenetic remodeling in the acquisition and maintenance of the CAF phenotype. We will systematically analyze how NNMT regulates the transcriptome and how alteration of epigenetics functionally drives the conversion of normal fibroblasts to CAFs. Since NNMT impinges on multiple metabolic pathways, in Aim II we will systematically assess the NNMT-driven metabolic state and its contribution to CAF differentiation and the promotion of tumor progression. Finally, in Aim III, our group will work with the NIH Center for Advancing Translational Sciences (NCATS) Chemical Genomics Center (NCGC) to discover compounds that inhibit NNMT biochemical activity, using state of the art high-throughput screening with an optimized NNMT biochemical screen of over 300,000 potential inhibitors. Lead compounds will be functionally screened in high- throughput 3D models of the tumor microenvironment at U of C. Successful inhibition of NNMT activity in the tumor stroma could result in a novel and clinically relevant approach to the treatment of metastatic ovarian cancer.

抽象的 浆液性上皮性卵巢癌 (OvCa) 通常表现为癌细胞在体内广泛扩散。 腹腔和显着的肿瘤负荷。 OvCa 转移瘤具有高比例的间质， 主要由癌症相关成纤维细胞 (CAF) 组成，这是一种已知可促进癌症相关的间充质细胞类型 肿瘤细胞的侵袭和转移。然而，目前尚不清楚正常成纤维细胞如何被重编程为 CAF 以及 CAF 如何促进肿瘤生长。为了直接解决这些悬而未决的问题，我们执行了 对 OvCa 患者样本中的原发性和转移性基质进行蛋白质组学分析，以确定 差异表达。我们检测到，特别是在转移瘤中，与相关的保守基质特征 转移包括烟酰胺 N-甲基转移酶 (NNMT) 的高基质表达。神经网络MT 催化甲基从 S-腺苷甲硫氨酸 (SAM) 转移到烟酰胺。这耗尽了 细胞 SAM 储存并导致组蛋白整体低甲基化和促肿瘤基因的表达。 在初步实验中，我们发现NNMT可以将正常成纤维细胞重编程为CAF，并且NNMT CAF 的抑制可阻止 OvCa 癌细胞粘附、增殖和体内肿瘤生长。在我们的系统中， NNMT 表达特异性调节 H3K4 和 H3K27 的三甲基化。 NNMT表达分析 OvCa 患者显示，强烈的基质 NNMT 表达与不良预后显着相关。 基于这些数据，该应用的主要假设是正常人中 NNMT 的表达 成纤维细胞通过代谢介导的表观遗传改变将它们转化为 CAF。拟议的 实验将系统地表征 NNMT 驱动的表观遗传和代谢的贡献 正常成纤维细胞向 CAF 转化的变化。在目标 I 中，我们建议研究 NNMT 驱动的 CAF 表型的获得和维持中的表观遗传重塑。我们将系统地 分析 NNMT 如何调节转录组以及表观遗传学的改变如何在功能上驱动 正常成纤维细胞向 CAF 的转化。由于 NNMT 影响多种代谢途径，因此在 Aim II 中我们 将系统地评估 NNMT 驱动的代谢状态及其对 CAF 分化和 促进肿瘤进展。最后，在目标 III 中，我们的小组将与 NIH 推进中心合作 转化科学 (NCATS) 化学基因组中心 (NCGC) 发现抑制化合物 NNMT 生化活性，采用最先进的高通量筛选和优化的 NNMT 超过 300,000 种潜在抑制剂的生化筛选。先导化合物将在高功能上进行筛选 加州大学肿瘤微环境的吞吐量 3D 模型。成功抑制 NNMT 活性 肿瘤基质可能会产生一种新颖的、临床相关的方法来治疗转移性卵巢 癌症。