A Tissue Engineering Approach to Analyzing Host-Microbe Interactions in Cancer

分析癌症中宿主-微生物相互作用的组织工程方法

基本信息

批准号：
9980814
负责人：
Daniel J Slade
金额：
$ 19.63万
依托单位：
VIRGINIA POLYTECHNIC INST AND ST UNIV
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9980814
关键词：
3-Dimensional 3D Print Address Anaerobic Bacteria Animal Model Attenuated Automobile Driving Bacteria Biological Models Biomimetics Blood Blood Circulation Blood Vessels Brain Cancer Vaccines Cells ChIP-seq Chronic Disease Coculture Techniques Colon Colon Carcinoma Colonic Neoplasms Colorectal Cancer Complement Complex Culture Media Devices Disease Distant Endothelial Cells Engineering Epithelial Epithelial Cells Epithelium Exhibits Fusobacterium nucleatum Future Gene Deletion Genetic Transcription Goals Health Human Hypoxia In Vitro Infection Invaded Knowledge Liver Lung Malignant Neoplasms Mammalian Cell Methods Microbe Microfluidic Analytical Techniques Microfluidics Modeling Monitor Neoplasm Metastasis Oncogenic Oral cavity Organ Organoids Paracrine Communication Pathway interactions Perivascular Neoplasm Phenotype Physiological Play Positioning Attribute Process Regulation Research Resolution Role Signal Transduction Site Sterility Study models System Technology Testing Tissue Engineering Tissue Model Tissues Tropism Validation bacterial genetics base biological systems cancer cell cancer prevention cancer therapy cell transformation colorectal cancer progression comparative cost effectiveness confocal imaging cost effective cost effectiveness epigenome epigenomics epithelial to mesenchymal transition experience genetic technology host-microbe interactions in vivo innovation migration neoplastic cell novel oral anaerobes pathogen recruit screening single-cell RNA sequencing spatiotemporal stem cell niche stem cells tool transcriptome transcriptomics tumor tumor immunology tumor microenvironment

项目摘要

There remains a critical knowledge gap regarding the spatiotemporal dynamics of bacterial tissue dissemination, as well as how a diverse range of bacteria broadly and differentially influence phenotypes referred to as the hallmarks of cancer. As one particularly illustrative example, Fusobacterium nucleatum is a resident, non-motile, Gram-negative anaerobe of the oral cavity with the ability to disseminate and cause lethal infections of the brain, lungs, and liver, and is often found in abundance in colorectal cancer (CRC) tissue. It is known that entry of this bacterium into host cells is a critical component in the activation of pro- oncogenic pathways, yet the tissue dissemination mechanisms for F. nucleatum past initial cellular entry in the oral cavity remains poorly understood. Recently it was demonstrated that this bacterium is able to survive within infected host tumor cells for long enough to be carried along to distant metastatic sites such as the liver. And we have more recently demonstrated that these bacteria invade CRC organoids, and intriguingly localize to crypt niches of critical importance in CRC progression. These observations suggest a potential role for bacteria in CRC and provide a rationale to further understand how F. nucleatum gains access to normally sterile tissues. We believe that recent breakthroughs in tissue engineering will enable studies that are capable of revealing the broad-ranging effects of bacteria in the tumor microenvironment (TME), including but not limited to i) the dissemination dynamics through blood vessels and within the tumor perivascular niche, and ii) the impacts of bacteria on CRC progression via regulation of epithelial-to- mesenchymal transition (EMT) that has been implicated in the colon stem cell niche. In this highly exploratory R21 project we will overcome the major limitations to such studies, which include the over- proliferation of tumor and bacterial cells in vitro, and the limited spatiotemporal resolution in vivo. We will leverage tissue engineering combined with microfluidic analysis and bacterial genetic technologies to enable high-resolution studies of tumor-microbe interactions in an in vitro biological system modeled after the human TME. We will proceed with the following aims: 1) Develop a microfluidic spheroid-based tissue model capable of mimicking F. nucleatum tropism out of the circulation and into the colon TME, and 2) Perform unbiased transcriptome and epigenome profiling of epithelial pathway activation resulting from F. nucleatum infection under fully-defined CRC TME conditions. More broadly, this project could result in an adaptable technology with higher throughput, accuracy, and cost-effectiveness compared with competing methods to analyze dynamic host-pathogen interactions, help to define new roles for tumor-microbe interactions in complex cancer processes, and aid in the identification and screening of future targets for cancer vaccines or therapies.

关于细菌组织的时空动力学仍然存在关键的知识差距传播，以及各种细菌如何广泛和差异化表型被称为癌症的标志。作为一个特别说明性的例子，核细菌核是口腔的居民，非运动，革兰氏阴性的厌氧菌，能够传播和引起大脑，肺部和肝脏的致命感染，通常在大肠癌中发现丰富（CRC）组织。众所周知，该细菌进入宿主细胞是激活促进细胞的关键成分致癌途径，然而，F.核的组织传播机制在初始细胞进入中口腔知之甚少。最近证明了该细菌能够在感染的宿主肿瘤细胞中生存足够长的时间，可以沿着遥远的转移部位携带作为肝脏。而且我们最近证明了这些细菌侵入CRC器官，并且有趣的是，在CRC进展中具有至关重要的地下个性属性。这些观察表明细菌在CRC中的潜在作用，并提供了一个理由，以进一步了解F. nutleatum如何增长进入正常无菌组织。我们认为，最近的组织工程突破将实现能够揭示细菌在肿瘤微环境中的广泛影响的研究（TME），包括但不限于i）通过血管和肿瘤内的传播动力学血管周期和II）细菌对CRC进展的影响通过调节上皮到 - 与结肠干细胞生态位有关的间充质转变（EMT）。在这方面探索性R21项目，我们将克服此类研究的主要局限性，其中包括过度体外肿瘤和细菌细胞的增殖，体内有限的时空分辨率。我们将利用组织工程与微流体分析和细菌遗传技术结合到启用对以以后建模的体外生物学系统中肿瘤微生物相互作用的高分辨率研究人类TME。我们将继续以下目的：1）开发基于微流体的组织能够模仿循环中并进入结肠TME的F. tupleatum tropism的模型，2）对F。在完全定义的CRC TME条件下的核感染。更广泛地，这个项目可能导致与竞争相比，具有更高吞吐量，准确性和成本效益的适应性技术分析动态宿主 - 病原体相互作用的方法，有助于定义肿瘤微叶的新作用复杂癌症过程中的相互作用，并有助于识别和筛选未来目标癌症疫苗或疗法。