Design and characterization of bacterial population dynamics in solid tumor models

实体瘤模型中细菌种群动态的设计和表征

基本信息

批准号：
10212134
负责人：
JEFF M HASTY
金额：
$ 64.29万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10212134
关键词：
Affect Ally Anaerobic Bacteria Animal Model Bacteria Bacterial Genes Behavior Biological Models Biosensing Techniques Biosensor CCL21 gene CRISPR/Cas technology Cancer Model Colonoscopy Colorectal Colorectal Cancer Colorectal Neoplasms Complex Cues Cytolysis Data Development Disease Drug Delivery Systems Engineered Probiotics Engineering Environment Escherichia coli Future Gases Gel Gene Cluster Gene Expression Genes Genetic Genome Glucose Goals Growth Growth Factor Harvest Health High Fat Diet Histology Human Human body Hypoxia Image Imaging technology Immunologic Surveillance In Situ Hybridization In Vitro Intercellular Fluid Intestines Lead Liquid substance Liver Luciferases Maintenance Malignant Neoplasms Malignant neoplasm of gallbladder Malignant neoplasm of liver Malignant neoplasm of lung Malignant neoplasm of ovary Malignant neoplasm of pancreas Measures Methods Microbe Microfluidic Microchips Microscopy Modeling Molecular Monitor Mucous Membrane Mus Nutrient Optical reporter Oral Organoids Pathologic Penetration Play Population Population Dynamics Primary carcinoma of the liver cells Probiotics Property Reporting Research Resolution Role Safety Solid Neoplasm Specificity Structure System Techniques Technology Testing Time Tissues Ultrasonography Vesicle base clinically relevant design design and construction drug production effectiveness study environmental change genome wide screen genome-wide high resolution imaging human disease improved in vivo in vivo Model luminescence malignant breast neoplasm malignant mouth neoplasm microbial microfluidic technology mouse model neoplastic nonalcoholic steatohepatitis novel pre-clinical preference promoter quorum sensing response screening synthetic biology tool tumor tumor growth tumor microenvironment

Affect Ally Anaerobic Bacteria Animal Model Bacteria Bacterial Genes Behavior Biological Models Biosensing Techniques Biosensor CCL21 gene CRISPR/Cas technology Cancer Model Colonoscopy Colorectal Colorectal Cancer Colorectal Neoplasms Complex Cues Cytolysis Data Development Disease Drug Delivery Systems Engineered Probiotics Engineering Environment Escherichia coli Future Gases Gel Gene Cluster Gene Expression Genes Genetic Genome Glucose Goals Growth Growth Factor Harvest Health High Fat Diet Histology Human Human body Hypoxia Image Imaging technology Immunologic Surveillance In Situ Hybridization In Vitro Intercellular Fluid Intestines Lead Liquid substance Liver Luciferases Maintenance Malignant Neoplasms Malignant neoplasm of gallbladder Malignant neoplasm of liver Malignant neoplasm of lung Malignant neoplasm of ovary Malignant neoplasm of pancreas Measures Methods Microbe Microfluidic Microchips Microscopy Modeling Molecular Monitor Mucous Membrane Mus Nutrient Optical reporter Oral Organoids Pathologic Penetration Play Population Population Dynamics Primary carcinoma of the liver cells Probiotics Property Reporting Research Resolution Role Safety Solid Neoplasm Specificity Structure System Techniques Technology Testing Time Tissues Ultrasonography Vesicle base clinically relevant design design and construction drug production effectiveness study environmental change genome wide screen genome-wide high resolution imaging human disease improved in vivo in vivo Model luminescence malignant breast neoplasm malignant mouth neoplasm microbial microfluidic technology mouse model neoplastic nonalcoholic steatohepatitis novel pre-clinical preference promoter quorum sensing response screening synthetic biology tool tumor tumor growth tumor microenvironment

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项目摘要

Project Summary It is increasingly clear that bacteria play an important role in human health. While it is natural to focus on how intestinal bacteria affect disease, intriguing ﬁndings have elucidated the extent to which bacteria inhabit solid tumors. Microbes have been detected in lung, pancreatic, breast, oral, gallbladder, ovarian, liver, and colorectal cancers. Localization has been ascribed to several mechanisms, including preference for anaerobic or facultative anaerobic bacteria to grow in the hypoxic core of tumors, presence of bacterial nutrients, lack of immune surveil- lance, and leakiness of the often poorly structured vasculature surrounding neoplastic tissue. This tendency for localization to solid tumors suggests that bacteria could be engineered for precise and robust drug production and delivery from within the solid tumor environment. This dovetails with 20 years of progress in synthetic biology, which has tended to focus on microbial engineering. However, information on how the tumor microenvironment affects bacterial growth is largely unknown. The microenvironment will affect bacterial gene expression that ul- timately underlies the functionality of engineered therapies, and it is difﬁcult to imagine a predictive framework for engineered bacterial therapies without a quantitative understanding of how bacteria react to the environment of a growing tumor. We will use a probiotic strain of E. coli with an established safety record to develop a novel class of biosensors to noninvasively investigate bacterial growth in the tumor microenvironment. Initially, we will develop lysis-based biosensors that respond to speciﬁc tumor environment targets: hypoxia, pH, glucose, and lactate (Aim 1). We will also engineer an inducible quorum sensing (QS) system that enables external control of bacterial population dynamics, including the ability to eliminate a speciﬁc strain whenever desired (Aim 1). Together these strains will allow us to modulate and monitor population dynamics in vivo, enabling both sens- ing of the local environment and maintenance of an external control switch. We will test these strains using an established in vitro organoid model (Aim 2) and in two clinically relevant animal models for solid tumor growth. Additionally, we will use our previously developed dynOMICS technology to screen tumor extract from the two animal models and construct a second suite of biosensors for monitoring the tumor environment (Aim 2). These biosensors will then be tested in the animal models. We will visualize bacterial populations in a colorectal tumor model with bacteria that are engineered to produce luciferase in order to monitor colony dynamics using our es- tablished methods (Aim 3). We will also build on recently reported technology whereby bacteria are modiﬁed for use with ultrasound through addition of gas vesicles that permit high resolution imaging of the engineered bac- teria. We will use the ultrasound method to investigate NASH-induced hepatocellular carcinoma (HCC) where a high-fat diet is used to induce HCC at 20 weeks in mice (Aim 4). This project will quantitatively characterize how bacterial strains sense, respond, and grow in the tumors. The results will establish a platform for future exploration of therapies that are produced and delivered by bacteria that grow within solid tumors.

项目摘要越来越明显的是，细菌在人类健康中起着重要作用。虽然很自然地专注于肠道细菌影响疾病，有趣的发现已经阐明了细菌居住在多大程度上在肺，胰腺，乳房，口服，胆囊，卵巢，肝脏和结直肠癌中检测到微生物癌症。本地化已分配给多种机制，包括偏爱厌氧或兼职厌氧菌细菌可在肿瘤的低氧核心中生长，存在细菌营养素，缺乏免疫外星长矛，以及肿瘤组织周围结构较差的脉管系统的泄漏。这种趋势对实体瘤的本地化表明，细菌可以用于精确和稳健的药物生产和从实体瘤环境中传递。这在合成生物学方面取得了20年的进展，它倾向于专注于微生物工程。但是，有关肿瘤微环境如何的信息影响细菌生长在很大程度上未知。微环境将影响细菌基因表达特别是，工程疗法的功能是基础，很难想象一个预测框架用于工程细菌疗法，没有对细菌如何对环境反应的定量了解肿瘤的生长。我们将使用具有既定安全记录的大肠杆菌的益生菌菌株来发展新颖生物传感器类别非侵入性研究肿瘤微环境中细菌的生长。最初，我们将开发基于裂解的生物传感器，以应对特定的肿瘤环境目标：缺氧，pH，葡萄糖，并裂开（目标1）。我们还将设计一个可诱导的法定感应（QS）系统，该系统启用外部控制细菌种群动力学，包括每当需要消除特定菌株的能力（AIM 1）。这些菌株一起将使我们能够在体内调节和监测种群动态，从而使两个传感器 - 当地环境和外部控制开关的维护。我们将使用一个建立的体外器官模型（AIM 2）和在两个临床相关的实体瘤生长模型中。此外，我们将使用先前开发的Dynomics技术从两者中筛选肿瘤提取物动物模型并构建了第二套生物传感器，用于监测肿瘤环境（AIM 2）。这些然后将在动物模型中测试生物传感器。我们将可视化结直肠肿瘤中的细菌种群具有生产荧光素酶的细菌模型，以便使用我们的ES- 提出的方法（AIM 3）。我们还将基于最近报道的技术，从而修饰细菌通过添加允许高分辨率成像的加油蔬菜与超声检查特里亚。我们将使用超声方法研究NASH诱导的肝细胞癌（HCC）高脂饮食用于在小鼠20周时诱导HCC（AIM 4）。该项目将定量特征细菌如何在肿瘤中感知，反应和生长。结果将建立一个未来的平台探索在实体瘤内生长的细菌产生和输送的疗法。