Micro-engineered capsules for spatial sampling of microbiome in vivo

摘要 背景和原理：我们建议进行探索性研究，以开发和测试微工程 可食用胶囊，用于采样胃肠道(GI)的内容物。已知的胃肠道 港口丰富多样的微生物群落(微生物群或微生物群)适应不同的酸度， 氧分压、胆汁浓度等条件存在于不同的器官中。这些微生物完成了 对消化和控制宿主的免疫功能至关重要的重要代谢功能。穿过 代谢产物的产生，这些微生物群落调节免疫反应，促进 上皮细胞的完整性。已知异常的(非生物的)微生物群与某些 炎症和对肠道感染抵抗力减弱等情况。高通量测序和 其他高级分析技术支持对复杂环境进行详细分析，如GI中所示 一条小路。与高通量测序和其他组学技术的快速发展相比，我们的 对胃肠道进行非侵入性采样的能力并不令人满意。大多数研究使用的生物分子都是从 以推断小肠和大肠的生态，并检测肠道病原体。我们正在提议 开发微型工程可食用胶囊，旨在从胃肠道的特定位置进行采样。 采样将使用由石蜡制成的无线触发的热驱动微型阀进行控制。 采样将使用嵌入式被动渗透泵，采样率为 100~300微米L/小时，不需要电源。外部磁性传感器阵列将使跟踪 胶囊由于蠕动而通过胃肠道。 具体目标：本研究有两个具体目标：1)体外和体外微球的设计、制造和测试 经过改造的可食用胶囊。体外试验将在断奶后解剖的肠道器官中进行。 猪；2)利用猪作为人胃肠道的模型，胶囊设计满足体外和体外的特异性 性能标准将在体内进行测试。核糖体RNA(16S)高通量测序将用于 描述胶囊采样的细菌群落。将采样的微生物区系的组成与 从猪肠不同部分收集的安乐死后的数据将揭示胶囊的能力 采集特定胃肠道器官的含量。 创新：我们提出的研究旨在开发一种新的研究工具。如果成功，这项技术 这将为研究胃肠道的生理学及其对扰动的反应提供一种新的方法 由肠道感染或其他情况引发。这项技术将促进肠道内容物的分析 以非侵入性的方式从不同的器官收集。预计这一领域的进一步发展 技术将导致临床应用，提高诊断感染性和非感染性胃肠道疾病的能力 条件。 好了！

Summary Background and rationale: We are proposing exploratory research to develop and test microengineered ingestible capsules designed to sample the content of the gastro-intestinal (GI) tract. The GI tract is known to harbor abundant and diverse microbial communities (microbiome or microbiota) adapted to different acidity, oxygen tension, bile concentration and other conditions found in different organs. These microbes fulfill important metabolic functions essential to digestion and to controlling immune functions of the host. Through the production of metabolites, these microbial communities regulate the immune response and promote the integrity of the epithelium. Abnormal (dysbiotic) microbiomes are known to be associated with certain conditions such as inflammation and impaired resistance to enteric infections. High-throughput sequencing and other advanced analytical techniques enable detailed analyses of complex environments as found in the GI tract. In contrast to the rapid development of high-throughput sequencing and other omics technologies, our ability to non-invasively sample the GI tract is unsatisfactory. Most research uses biomolecules extracted from feces to infer the ecology of the small and large intestine and to detect enteric pathogens. We are proposing to develop microengineered ingestible capsules designed to sample from specific locations in the GI tract. Sampling will be controlled using wirelessly triggered thermally actuated microvalves made from paraffin wax. Sampling will be achieved using an embedded passive osmotic pump capable of sampling at a rate of 100~300 µL/hr without the need for electrical power. External magnetic sensor array will enable tracking of the capsule as it moves through the GI tract due to peristalsis. Specific Aims: The research has two Specific Aims; 1) Design, manufacture and test in vitro and ex vivo micro- engineered ingestible capsules. Ex vivo testing will be performed in intestinal organs dissected from weaned pigs; 2) Using the pig as a model of the human GI tract, capsule designs meeting specific in vitro and ex vivo performance criteria will be tested in vivo. Ribosomal RNA (16S) high-throughput sequencing will be used to profile the bacterial communities sampled by the capsule. Comparing the make-up of sampled microbiota with those collected post-euthanasia from different sections of the pig intestine will reveal the ability of the capsules to sample the content of specific GI organs. Innovation: The research we are proposing aims to develop a new research tool. If successful, the technology will enable a novel approach to studying the physiology of the GI tract and its response to perturbation triggered by enteric infections or other conditions. The technology will facilitate the analysis of gut content collected from various organs in a non-invasive manner. It is anticipated that further development of this technology will lead to clinical application with improved capabilities to diagnose infectious and non-infection GI conditions. !