Microbial Synthesis of Therapeutic Bile Acids for Alzheimer's Disease

微生物合成治疗阿尔茨海默病的胆汁酸

• 批准号: 10602316
• 负责人: Gunda I. Georg
• 金额: $ 134.42万
• 依托单位: METSELEX, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10602316
• 关键词: APP-PS1; Abate; Acute; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Amyloid beta-Protein; Anabolism; Animal Disease Models; Animal Model; Animals; Apoptosis Inhibitor; Apoptotic; Artemia; Bacteria; Bile Acids; Biological Assay; Biomanufacturing; Businesses; Cause of Death; Cell Death; Cell model; Cells; Cellular Membrane; Chemicals; Cholesterol; Cholic Acids; Chronic; Cognitive deficits; Complex; Contracts; DNA; DNA biosynthesis; Dendrites; Dendritic Spines; Development; Disease; Dose; Drug Kinetics; Economic Burden; Engineering; Enzymes; Ergosterol; Fermentation; Foundations; Gallbladder; Genes; Genetic; Goals; Grant; Harvest; Hippocampus; Human; In Vitro; Industrialization; Industry; Lead; Learning; Livestock; Marketing; Memory impairment; Metabolic; Metabolic Pathway; Minnesota; Mitochondria; Nerve Degeneration; Neurodegenerative Disorders; Neuroprotective Agents; Organism; Outcome; Pathology; Pathway interactions; Patient-Focused Outcomes; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacodynamics; Phase; Phase I Clinical Trials; Phosphorylation; Preparation; Process; Production; Productivity; Property; Proteins; Qualifying; Reactive Oxygen Species; Recombinants; Research; Role; Small Business Technology Transfer Research; Societies; Solid; Source; Specificity; Synapses; System; Techniques; Technology; Technology Transfer; Testing; Therapeutic; Treatment Cost; Universities; Ursodeoxycholic Acid; Vertebral column; Work; Yeasts; Zebrafish; amyloid precursor protein processing; analog; analytical method; care costs; combat; cost estimate; design; design and construction; disability; drug development; endoplasmic reticulum stress; experience; frontal lobe; gene product; gut microbiome; improved; in vitro Assay; in vivo; in vivo evaluation; large scale production; manufacture; manufacturing organization; manufacturing technology; microbial; microbial host; mouse model; neural; neuroprotection; new technology; novel; novel therapeutics; pharmacologic; potency testing; prevent; reconstitution; response; scale up; screening; success; supply chain; tau Proteins; tauroursodeoxycholic acid; tool; unethical

ABSTRACT This Phase STTR Phase II proposal aims to engineer and scale-up a synthetic metabolic pathway in a microbial host to produce UDCA and related compounds. Additionally, the UDCA produced from the engineered synthetic metabolic pathway will be used to synthesize derivatives for testing in our Alzheimer’s Disease cell and animal models. The proposed work has high intellectual merit for the following reasons: (i) UDCA biosynthesis does not occur in any organism whose cultivation can be scaled to meet global demand, (ii) UDCA requires a multi- organism biosynthetic pathway (animal cholic acids converted to UDCA by the gut microbiome) that will have to be reconstituted in a single cell, and (iii) the anticipated scale and complexity of this engineering effort (combining more than a dozen genes from four organisms) are on the edge of what is feasible. Technical hurdles involve the discovery of new enzymes that convert ergosterol to cholesterol (a process known to exist in brine shrimp), achieving proper localization and enzymatic activity for fifteen recombinant gene products in yeast, and balancing the expression levels of each gene to support high-titer production of UDCA in yeast. The combined Metselex/University of Minnesota team will overcome these hurdles using their platform for multi-gene pathway refactoring and high-throughput DNA assembly and analytical methods. Experience with these tools/approaches and success in engineering similar synthetic metabolic pathways qualifies this team conduct these studies and will enable them to accomplish their Phase II goals.

摘要 该阶段STTR第二阶段提案旨在设计和扩大微生物中的合成代谢途径， 生产UDCA和相关化合物的宿主。此外，由工程合成的UDCA 代谢途径将用于合成衍生物，用于在我们的阿尔茨海默病细胞和动物中进行测试 模型建议的工作具有很高的智力价值，原因如下：（i）UDCA生物合成确实 不会发生在任何生物体，其培养可以扩大规模，以满足全球需求，（ii）UDCA需要多- 生物体生物合成途径（动物胆酸通过肠道微生物组转化为UDCA）， 在单个细胞中重建，以及（iii）这种工程努力的预期规模和复杂性（组合 来自四种生物体的十几个基因）正处于可行的边缘。技术障碍包括 发现了将麦角甾醇转化为胆固醇的新酶（已知存在于卤虫中的过程）， 对15个重组基因产物在酵母中进行了正确的定位和酶活性测定， 每个基因的表达水平，以支持酵母中UDCA的高滴度生产。将合并的 Metselex/明尼苏达大学团队将利用他们的多基因途径平台克服这些障碍 重构和高通量DNA组装和分析方法。使用这些工具/方法的经验 在设计类似的合成代谢途径方面的成功使这个团队有资格进行这些研究， 这将有助于他们实现第二阶段的目标。