Engineering probiotics for tuberculosis therapy

用于结核病治疗的工程益生菌

• 批准号: 10511520
• 负责人: Tal Danino
• 金额: $ 28.97万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10511520
• 关键词: 3-Dimensional; Antibiotics; Antimycobacterial Agents; Automobile Driving; Bacteria; Biological Assay; Cells; Cessation of life; Coculture Techniques; Combined Antibiotics; Complement; Coupling; Cutaneous; Data; Disease; Drug Delivery Systems; Drug resistance; Engineered Probiotics; Engineering; Environment; Escherichia coli; Euthanasia; Genetic; Genetic Engineering; Genetic Programming; Genus Mycobacterium; Granuloma; Granulomatous; Granulomatous disease; Growth; Histologic; Home; Hypoxia; Immune response; In Vitro; Individual; Infection; Infiltration; Intelligence; Libraries; Measures; Medicine; Microbe; Microscopy; Modeling; Molecular; Monitor; Mus; Mycobacterium Infections; Mycobacterium marinum; Mycobacterium tuberculosis; Necrosis; Organ; Pathogenicity; Pathologic; Peptides; Persons; Probiotics; Production; Proteins; Recombinants; Reporter; Reporter Genes; Resistance; Safety; Site; Solid Neoplasm; Specificity; T-Lymphocyte; Techniques; Testing; Therapeutic; Therapeutic Agents; Time; Tissues; Toxic effect; Treatment Efficacy; Tuberculosis; Virulent; Work; antimicrobial; base; cancer therapy; clinically relevant; delivery vehicle; density; design; draining lymph node; in vivo; in vivo imaging system; interest; macrophage; mouse model; mycobacterial; neutrophil; novel; novel strategies; pathogen; safety testing; small molecule; synergism; synthetic biology; therapeutic candidate; therapeutic evaluation; therapeutic protein; tool; treatment duration; tuberculosis granuloma; tuberculosis treatment; tumor; tumor microenvironment; two-dimensional

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.

项目摘要 活细胞和微生物的工程正在推动医学的新时代。这种变革性的方法 允许活细胞的遗传编程智能地感知和响应健康和患病的环境， 身体里的骨头。细菌特别产生了显着的最近的兴趣，由于其遗传tracta- 渗透疾病部位的能力。大量研究表明，细菌递送治疗剂 有效载荷选择性地进入肿瘤核心，证明了特异性和有效性，这是用其他方法无法达到的。 分子疗法鉴于癌症治疗的这种模式，细菌提供了一个独特的机会， 被设计成智能运输工具，运送到难以到达的疾病地点。 我们最近发现益生菌E.大肠杆菌Nissle 1917（EcN）选择性地归巢于肉芽肿，病理性 在包括结核病在内的感染部位形成的区域。结核病每年造成150万人死亡， 常规抗生素受到毒性、疗程长和耐药性的限制。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， 支持选择性细菌定植的类似于肿瘤的组织。 该提案的目的是设计EcN，使其成为肉芽肿的家园，并在当地生产治疗药物， 消灭致病性结核分枝杆菌。我们将使用合成生物学方法进行基因工程- neer EcN as a intelligent智能drug药物delivery交付vehicle车辆.我们将利用缺氧感应生长回路， 益生菌生长的缺氧，坏死肉芽肿环境。使用新的体外3D共培养试验，我们将 筛选抗微生物蛋白文库以鉴定最有效的抗分枝杆菌治疗候选物。 我们将测试工程E.大肠杆菌中进行。这 治疗肉芽肿病的一种全新方法将为利用工程菌提供机会 作为肉芽肿感染的治疗剂。