Creating a niche for engineered live biotherapeutics

项目总结 这项提议的长期目标是改善小分子输送的结果， 通过工程活的生物治疗细菌将多肽和蛋白质输送到人体肠道。至 要做到这一点，我们将使用两种策略来优化肠道内的生物治疗生长 环境。首先，我们将匹配益生菌化合物(未被消化的复杂碳水化合物 身体)异源表达的酶，使细胞能够利用这些特权 碳源。其次，我们将在我们的候选生物疗法中创建基因组规模的文库， E.ColiNissle 1917和乳酸乳球菌，以确定增加人体健康的基因产品 肠胃和肠胃一样的环境。这些方法将在体外培养、小肠 有机化合物共培养，并在体内测定对小鼠肠道的浓缩生物治疗作用 群体，提高治疗性小分子、多肽或蛋白质的生产 兴趣，以及与人类肠道越来越逼真的系统的表型结果。

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.