Programmable Probiotics for Cancer

可编程益生菌治疗癌症

• 批准号: 9189187
• 负责人: Tal Danino
• 金额: $ 24.9万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9189187
• 关键词: Address; Adenocarcinoma; Apoptotic; Bacteria; Bacterial Luciferases; Behavior; Binding; Biological Markers; Cancer Model; Cell Line; Cells; Cleaved cell; Colorectal; Colorectal Cancer; Computer Simulation; Cytolysis; Data; Detection; Development; Diagnosis; Diagnostic; Dimensions; Dose-Limiting; Engineering; Environment; Enzyme-Linked Immunosorbent Assay; Fluorescence; Genes; Genetic; Genetically Engineered Mouse; Growth; Health; Histocompatibility Testing; Homing; Hour; Human; Human body; Image; Immune response; Immunocompetent; In Vitro; Infection; Injectable; Intestines; Laboratories; Luciferases; MDA MB 435; Malignant Neoplasms; Measurement; Measures; Membrane Proteins; Mentors; Metastatic Neoplasm to the Liver; Methods; Microbe; Microfluidic Microchips; Microfluidics; Microscopy; Modeling; Monitor; Mus; Necrosis; Oral; Oral Administration; Oxygen; Paper; Peptides; Pharmaceutical Preparations; Phase; Polyps; Population; Population Density; Prevalence; Probiotics; Production; Research; Ribosomes; Science; Sensitivity and Specificity; Specificity; Staging; Surface; Technology; Testing; Therapeutic; Time; Tissues; Toxic effect; Translating; Translations; Urine; Weight; base; clinical application; clinically relevant; design; dosage; engineering design; imaging system; improved; in vivo; in vivo imaging; intravenous administration; mathematical model; microbial; microbiome; mouse model; optimism; pathogen; peptide drug; pre-clinical; promoter; receptor; subcutaneous; synthetic biology; targeted treatment; time use; translational medicine; tumor; tumor progression; uptake; vector

 DESCRIPTION (provided by applicant): Over the past decade, the long-held view of bacteria as pathogens has been transformed by microbiome data revealing an astounding prevalence of microbes within the human body. These bacteria exist not only in well- known regions such as the intestines, but also in a multitude of tissue types including tumors. Given this prevalence, microbes represent a natural platform for the development of diagnostics and therapies engineered to sense their environment and deliver drugs to tumors. Current & Mentored Research: Our preliminary results have shown that oral delivery of probiotic bacteria can colonize colorectal liver metastases and enzymatically cleave an injected substrate that can be detected in the urine (Danino et al., Science Translational Medicine, in revision). Building upon this result to improve sensitivity and specificity, we will engineer a gene circuit to lyse at a threshold population density within the tumor environment and release a urine diagnostic peptide. Circuit parameters will be predicted by mathematical modeling, characterized by microfluidic measurements, and experimentally tuned by genetic copy number and ribosome binding strengths to optimize for detection of the diagnostic peptide in mouse models. Independent Phase Research: Following initial characterization of the engineered diagnostic circuit, we will assess colonization in more realistic orthotopic and genetically engineered mouse models of colorectal cancer. We will determine earliest stages at which bacteria colonization occurs and simultaneously measure diagnostic peptide concentrations in the urine. We will target our probiotics with tumor-homing peptides, express pro- apoptotic peptides as therapeutics, and monitor progression of tumors as a function of time. Altogether, the programmable probiotic platform will allow for more sophisticated microbial diagnostics and therapeutics for cancer and establish an engineering framework for in vivo applications for synthetic biology.

 描述（由申请人提供）：在过去的十年中，长期以来将细菌视为病原体的观点已经被微生物组数据所改变，这些数据揭示了人体内微生物的惊人流行。这些细菌不仅存在于肠道等众所周知的区域，而且存在于包括肿瘤在内的多种组织类型中。鉴于这种普遍性，微生物代表了开发诊断和治疗的天然平台，这些诊断和治疗旨在感知其环境并向肿瘤提供药物。当前和指导研究：我们的初步结果已经表明，益生菌的口服递送可以定殖结肠直肠肝转移并酶促切割可以在尿中检测到的注射底物（Danino等人，科学转化医学，修订版）。基于这一结果，以提高灵敏度和特异性，我们将设计一个基因电路，以在肿瘤环境中的阈值群体密度下裂解，并释放尿液诊断肽。回路参数将通过数学建模来预测，其特征在于微流体测量，并通过遗传拷贝数和核糖体结合强度进行实验调整，以优化小鼠模型中诊断肽的检测。独立阶段研究：在工程诊断电路的初步表征之后，我们将评估更现实的原位和基因工程小鼠结直肠癌模型中的定植。我们将确定细菌定植发生的最早阶段，同时测量尿液中的诊断肽浓度。我们将用肿瘤归巢肽靶向我们的益生菌，表达促凋亡肽作为治疗剂，并监测肿瘤随时间的进展。总之，可编程益生菌平台将允许更复杂的微生物诊断和癌症治疗，并为合成生物学的体内应用建立工程框架。