Engineering probiotics for tuberculosis therapy

项目总结 活细胞和微生物的工程学正在推动医学的新时代。这种变革性的方法 使活细胞的遗传编程能够智能地感知和响应健康和患病的环境-- 身体中的隆起物。细菌因其遗传域而引起了人们的极大兴趣-- 能力和能力渗透到疾病部位。大量研究表明，细菌传递的治疗作用 有效载荷选择性地进入肿瘤核心，显示出其他方法无法达到的特异性和有效性 基于分子的治疗学。鉴于癌症治疗中的这一范例，细菌提供了一个独特的机会 被设计成智能运送工具，到达难以到达的疾病地点。 我们最近发现，益生菌E.coliNissle 1917(ECN)选择性地引起肉芽肿，病理性 在包括结核病在内的感染地点发展起来的区域。结核病每年导致150万人死亡，但 常规抗生素具有毒性大、疗程长、抗药性强等特点。重要的是，虽然 传统疗法难以达到，肉芽肿具有独特的坏死和缺氧微环境- 类似于支持选择性细菌定植的肿瘤。 这项建议目标是使ECN成为肉芽肿的家园，并在当地生产治疗药物 消除致病结核分枝杆菌。我们将使用合成生物学方法进行基因工程-- Neer ECN作为一种智能药物输送工具。我们将利用低氧感应生长电路进一步限制 益生菌生长在缺氧、坏死性肉芽肿环境中。使用一种新的体外3D共培养实验，我们将 筛选抗菌蛋白文库，以确定最有效的抗分枝杆菌治疗候选药物。 我们将使用分枝杆菌肉芽肿小鼠模型来测试工程大肠杆菌的治疗效果。这 从根本上说，治疗肉芽肿性疾病的新方法将为利用工程菌打开机会 作为肉芽肿性感染的治疗剂。

PROJECT SUMMARY The engineering of living cells and microbes is driving a new era of medicine. This transformative approach allows for the genetic programming of living cells to intelligently sense and respond healthy and diseased envi- ronments in the body. Bacteria have specifically generated significant recent interest due to their genetic tracta- bility and ability to infiltrate disease sites. A multitude of studies have shown bacterial delivery of therapeutic payloads selectively into tumor cores, demonstrating specificity and efficacy that is otherwise unattainable with molecular-based therapeutics. Given this paradigm in cancer therapy, bacteria present a unique opportunity to be engineered as intelligent delivery vehicles to inaccessible disease sites. We recently discovered that probiotic E. coli Nissle 1917 (EcN) selectively home to granulomas, pathological regions developed at infection sites including tuberculosis. Tuberculosis kills 1.5 million people each year but conventional antibiotics are limited by toxicity, long treatment courses, and drug resistance. Importantly, while difficult for conventional therapeutics to reach, granulomas possess unique necrotic and hypoxic microenviron- ment similar to tumors that supports selective bacterial colonization. The objective of this proposal is to engineer EcN to home to granulomas and locally produce therapeutics to eliminate pathogenic Mycobacterium tuberculosis. We will use synthetic biology approaches to genetically engi- neer EcN as an intelligent drug delivery vehicle. We will utilize hypoxia-sensing growth circuits that further restrict probiotic growth the hypoxic, necrotic granuloma environment. Using a novel in vitro 3D coculture assay, we will screen a library of antimicrobial proteins to identify the most potent antimycobacterial therapeutic candidates. We will test the therapeutic efficacy of engineered E. coli using mycobacterium granuloma mouse models. This fundamentally new approach to granulomatous disease will open up opportunities to utilize engineered bacteria as therapeutic agent for granulomatous infections.