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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos s disease model Alzheimer&apos 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

项目摘要

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.

抽象的此阶段STTR II期建议旨在在微生物中设计和扩大合成代谢途径宿主生产UDCA和相关化合物。此外，由工程合成产生的UDCA 代谢途径将用于合成在阿尔茨海默氏病细胞和动物中测试的衍生物型号。拟议的工作具有以下原因具有很高的智力优点：（i）UDCA生物合成确实（ii）UDCA不得有机体生物合成途径（由肠道微生物组转化为UDCA的动物胆酸）将不得不在单个单元格中重构，（iii）此工程工作的预期规模和复杂性（结合）来自四个生物体的十几个基因都处于可行的边缘。技术障碍涉及发现将麦角固醇转化为胆固醇的新酶（已知存在于盐水虾中的过程），在酵母中实现15种重组基因产物的适当定位和酶活性，并平衡每个基因的表达水平支持酵母中UDCA的高电位产生。组合 Metselex/明尼苏达大学团队将使用其多基因途径的平台克服这些障碍重构和高通量DNA组装和分析方法。这些工具/方法的经验和工程的成功类似合成代谢途径符合该团队进行这些研究的资格，并将使他们能够实现第二阶段的目标。