Micro-engineered capsules for spatial sampling of microbiome in vivo

总结 背景和基本原理：我们正在提出探索性研究，以开发和测试微工程 设计用于对胃肠道（GI）内容物进行采样的可摄取胶囊。众所周知，胃肠道 拥有丰富多样的微生物群落（微生物组或微生物群），适应不同的酸度， 氧分压、胆汁浓度和不同器官中发现的其他条件。这些微生物满足了 对消化和控制宿主免疫功能至关重要的重要代谢功能。通过 代谢产物的产生，这些微生物群落调节免疫反应并促进免疫应答。 上皮的完整性。已知异常（生态失调）微生物组与某些 例如炎症和对肠道感染的抵抗力受损的病症。高通量测序和 其他先进的分析技术能够对地理信息系统中发现的复杂环境进行详细分析 道。与高通量测序和其他组学技术的快速发展相比，我们的 非侵入性地对胃肠道取样的能力是不令人满意的。大多数研究使用的生物分子是从 粪便来推断小肠和大肠的生态并检测肠道病原体。我们建议 开发微工程可摄取胶囊，旨在从胃肠道的特定位置采样。 将使用由石蜡制成的无线触发热致动微型阀控制采样。 将使用能够以以下速率采样的嵌入式被动渗透泵进行采样： 100~300微升/小时，无需电源。外部磁传感器阵列将能够跟踪 胶囊，因为它通过胃肠道移动由于消化道。 具体目的：本研究有两个具体目的：1）设计、制造和测试体外和离体微生物制剂。 工程可摄取胶囊。离体试验将在断奶仔猪解剖的肠器官中进行。 2）使用猪作为人胃肠道的模型，胶囊设计满足特定的体外和离体 将在体内测试性能标准。核糖体RNA（16 S）高通量测序将用于 对胶囊取样的细菌群落进行分析将采样微生物群的组成与 从猪肠的不同部分收集的那些将揭示胶囊的能力 对特定胃肠道器官进行取样 创新：我们提出的研究旨在开发一种新的研究工具。如果成功的话， 将使一种新的方法来研究胃肠道的生理学及其对扰动的反应 由肠道感染或其他情况引发。该技术将有助于分析肠道内容物 以非侵入性的方式从各种器官中收集。预计这方面的进一步发展 技术将导致临床应用，提高诊断感染性和非感染性GI能力 条件 !

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. !