Characterization of polyamine biosynthetic enzymes from human gut microbes associated with colon and pancreatic cancer

与结肠癌和胰腺癌相关的人类肠道微生物的多胺生物合成酶的表征

• 批准号: 10664019
• 负责人: Jeffrey S McFarlane
• 金额: $ 16.25万
• 依托单位: FORT LEWIS COLLEGE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10664019
• 关键词: Adenosylmethionine Decarboxylase; Amines; Anabolism; Aspartate; Bacterial Gene Expression Regulation; Bacteroides; Bacteroides fragilis; Biological Assay; Biological Process; Campylobacter; Carbon Dioxide; Carboxy-Lyases; Cell Proliferation; Cells; Clinical Trials; Clostridium; Colon Carcinoma; DNA Repair; Data; Decarboxylation; Development; Disease; Drug Design; Drug Targeting; Enzyme Kinetics; Enzymes; Future; Genes; Goals; Human; Kinetics; Life; Link; Malignant Neoplasms; Malignant neoplasm of pancreas; Metabolic Pathway; Microbial Biofilms; Minor; NADP; NADPH Dehydrogenase; Operon; Organism; Orthologous Gene; Oxidoreductase; Pathway interactions; Patients; Physical condensation; Polyamines; Process; Production; Public Health; Putrescine; Role; S-Adenosylhomocysteine; Spermidine; Spermidine Synthase; Structural Models; Structure; Variant; Vibrio; Work; X-Ray Crystallography; anti-cancer therapeutic; aspartic semialdehyde; cancer cell; colon cancer progression; colorectal cancer treatment; commensal bacteria; design; enzyme pathway; enzyme structure; gut bacteria; gut microbes; gut microbiome; inhibitor; kinetic model; member; new therapeutic target; norspermidine; programs; therapeutic target; tumorigenesis; uptake

PROJECT SUMMARY Polyamine biosynthesis in human cancer cells is a long-standing therapeutic target. Accumulating evidence suggests that polyamine production by gut microbes contributes to the progression of colon and pancreatic cancer. Biosynthesis of the polyamine spermidine by gut bacteria follows a pathway distinct from the human pathway making this bacterial pathway a unique drug target. The two central enzymes in this pathway, carboxyspermidine dehydrogenase and carboxyspermidine decarboxylase, have received only limited characterization. We propose to solve the first structure of a carboxyspermidine dehydrogenase and to development kinetic and structural models describing the function of both carboxyspermidine dehydrogenase and carboxyspermidine decarboxylase in major gut constituents from the Bacteroides and Clostridium genera. These aims serve as a necessary precursor to a future program of mechanism-guided drug design. The long- term goal of this work is to open new avenues for the treatment of colorectal and pancreatic cancer. Interestingly, the genes of predicted polyamine biosynthetic enzymes from many gut genera appear to encode redundant paths toward the production of spermidine. We hypothesize that these paths are not redundant, but generate unique polyamine products. We propose the use of LC-TQ-MS assays to delineate the polyamine products from apparently redundant enzyme pairs found in Clostridium. Spermidine synthase is a key focus of this aim because the prokaryotic orthologs of this enzyme have been shown to produce a variety of polyamine products besides spermidine. This work will expand our fundamental understanding of the functional variations found within a key gut microbe metabolic pathway: polyamine biosynthesis.

项目概要 人类癌细胞中的多胺生物合成是一个长期存在的治疗靶点。积累证据 表明肠道微生物产生的多胺有助于结肠和胰腺的进展 癌症。肠道细菌生物合成多胺亚精胺遵循与人类不同的途径 途径使该细菌途径成为独特的药物靶点。该途径中的两种中心酶， 羧基亚精胺脱氢酶和羧基亚精胺脱羧酶，仅收到有限的 表征。我们建议解决羧基亚精胺脱氢酶的第一个结构并 描述羧基亚精胺脱氢酶功能的发育动力学和结构模型 拟杆菌属和梭状芽胞杆菌属主要肠道成分中的羧基亚精胺脱羧酶。 这些目标是未来机制引导药物设计计划的必要先导。长- 这项工作的长期目标是为结直肠癌和胰腺癌的治疗开辟新途径。有趣的是， 来自许多肠道属的预测多胺生物合成酶的基因似乎编码冗余 生产亚精胺的途径。我们假设这些路径不是冗余的，而是生成 独特的多胺产品。我们建议使用 LC-TQ-MS 分析来描述来自 在梭菌中发现明显冗余的酶对。亚精胺合酶是这一目标的关键焦点，因为 该酶的原核直系同源物已被证明可以产生多种多胺产品，除了 亚精胺。这项工作将扩展我们对密钥中发现的功能变化的基本理解 肠道微生物代谢途径：多胺生物合成。