Creating a niche for engineered live biotherapeutics

为工程活生物治疗创造利基市场

• 批准号: 10622534
• 负责人: Thomas J Mansell
• 金额: $ 29万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10622534
• 关键词: Bacteria; Biological Response Modifier Therapy; Carbohydrates; Carbon; Cells; Coculture Techniques; Complex; Drug Delivery Systems; Engineering; Environment; Enzymes; Escherichia coli; Goals; Growth; Human; In Vitro; Injections; Lactococcus lactis; Libraries; Methods; Mus; Organoids; Outcome; Peptides; Pharmaceutical Preparations; Phenotype; Population; Production; Proteins; Small Intestines; Source; System; Testing; Therapeutic; delivery vehicle; disease diagnosis; fitness; gene product; genome-wide; improved outcome; in vivo; interest; prebiotics; public health relevance; small molecule; small molecule therapeutics

PROJECT SUMMARY The long-term goal of this proposal is to improve outcomes of delivery of small molecules, peptides, and proteins to the human gut by engineered live biotherapeutic bacteria. To accomplish this, we will use two strategies to optimize biotherapeutic growth in the gut environment. First, we will match prebiotic compounds (complex carbohydrates not digested by the body) with heterologous expression of enzymes that allow cells to utilize these privileged carbon sources. Second, we will create genome-scale libraries in our candidate biotherapeutics, E. coli Nissle 1917 and Lactococcus lactis, to identify gene products that increase fitness in the gut and gut-like environments. The methods will be tested in in vitro culture, small intestinal organoid co-culture, and in vivo in the mouse gut to determine enrichment of the biotherapeutic population, enhanced production of the therapeutic small molecule, peptide, or protein of interest, and phenotypic outcome in systems of increasing verisimilitude to the human gut.

项目概要 该提案的长期目标是改善小分子的递送结果， 通过工程化的活生物治疗细菌将肽和蛋白质输送到人体肠道。到 为了实现这一目标，我们将使用两种策略来优化生物治疗药物在肠道中的生长 环境。首先，我们将匹配益生元化合物（不被消化的复杂碳水化合物） 身体）具有酶的异源表达，使细胞能够利用这些特权 碳源。其次，我们将在候选生物治疗药物中创建基因组规模的文库， 大肠杆菌 Nissle 1917 和乳酸乳球菌，鉴定可增强身体健康的基因产物 肠道和类肠道环境。该方法将在体外培养、小肠 类器官共培养，并在小鼠肠道内进行体内测定，以确定生物治疗药物的富集 群体，增强治疗性小分子、肽或蛋白质的产生 人们对系统的兴趣和表型结果越来越接近人类肠道。