NCS-FO: Developing engineering solutions to investigate microbiome-to-neuron communication

NCS-FO：开发工程解决方案来研究微生物组与神经元的通讯

• 批准号: 1926793
• 负责人: Reza Ghodssi
• 金额: $ 100万
• 依托单位: University of Maryland, College Park
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2023-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1926793&HistoricalAwards=false
• 关键词: NCS; FO; Developing; engineering; solutions

The goal of this project is to create an engineering solution to measure and predict the molecular communication across the gut-microbiome-brain axis. This platform has the potential to facilitate the fundamental understanding of gut microbiome communication with the nervous system. The project will quantify release patterns of key molecules involved in this cross-talk and identify their influence on neural activation and behavior. The gut-microbiome-brain axis, comprising a vast network of nerves innervating the gut and propagating signals to the brain, is a major influencer of behavior and cognition. The neurotransmitter serotonin is a key molecule in this pathway; gut epithelial cells sense luminal conditions and release serotonin to stimulate nearby neurons. The gut microbiome has been shown to mediate this serotonin release, a process that is also linked to the co-occurrence of gastric and neural disorders. The technical underpinnings of this work involve designing and constructing a device that enables researchers to assemble the essential components of the gut-microbiome-neuron tissue interface. The device is fabricated with sensors to obtain information that is currently inaccessible - collecting molecular information at the length and time scales of the cells and tissues under investigation. The data extracted from this platform will enable temporal correlation and prediction of microbial, gut, and neural signaling patterns. This work provides opportunities to bring together researchers and stakeholders from various disciplines including electrical and computer engineering, bioengineering, molecular biology, neuroscience, and data science to develop a system-oriented approach. Further, this project promotes the participation of women, historically underrepresented in engineering, and undergraduates through programs such as Women in Engineering Research Fellowship and First-Year Innovation and Research Experience (FIRE). Multidisciplinary engineering methods are essential to building an in vitro discovery platform capable of directly monitoring chemical transduction patterns along the gut-neuron axis. In TASK 1, electrochemical sensors will be directly fabricated on a porous cell culture substrate, allowing direct access to cellular and molecular mechanisms of an in vitro model gut epithelium. Impedance monitoring of the cell layer will detect physical changes over time (e.g., barrier integrity). Potentiometric monitoring will detect real-time serotonin released from gut cells due to bacterial metabolite stimulation. In TASK 2, the neural effect of gut serotonin signaling will be studied by exposing this cell-released serotonin to an isolated ex vivo crayfish nerve cord with connected and innervated hindgut. Neurobehavioral activation patterns will be recorded during hindgut peristalsis in motor and sensory neurons that bidirectionally connect the central and enteric nervous systems. Machine learning approaches will identify key variables to quantify discrete serotonin release and neuronal activation patterns. In TASK 3, the mucosal layer of the gut epithelium will be colonized with specific gut microbes to assess bacterial influence on barrier integrity, serotonin release patterns, and resulting neuromuscular activation. Classification via machine learning will quantify the wholistic and synergistic effects of different microbial combinations on time-dependent serotonin release profiles and downstream effects. There are multiple novel aspects of this work. First, this is a new platform implementing extensive integrated cell-interfacial sensors for direct access to real-time cell and molecular data. Second, the use of this technology to investigate the interplay between gut and nervous system can give unprecedented insight into the vast and relatively inaccessible gut-brain transduction pathways. Third, machine learning analysis can identify meaningful patterns of serotonergic communication and predict the expected impact of gut bacteria on neural behavior.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该项目的目标是创建一个工程解决方案，以测量和预测肠道-微生物组-大脑轴上的分子通信。该平台有可能促进对肠道微生物组与神经系统通信的基本理解。该项目将量化参与这种串扰的关键分子的释放模式，并确定它们对神经激活和行为的影响。肠道-微生物组-大脑轴，包括支配肠道并将信号传播到大脑的巨大神经网络，是行为和认知的主要影响因素。神经递质5-羟色胺是这一通路中的关键分子;肠道上皮细胞感知管腔条件并释放5-羟色胺以刺激附近的神经元。肠道微生物组已被证明介导这种血清素释放，这一过程也与胃和神经疾病的共同发生有关。这项工作的技术基础涉及设计和构建一种设备，使研究人员能够组装肠道-微生物组-神经元组织界面的基本组成部分。该设备与传感器一起制造，以获得目前无法获得的信息-收集正在研究的细胞和组织的长度和时间尺度上的分子信息。从该平台提取的数据将使微生物，肠道和神经信号模式的时间相关性和预测成为可能。这项工作提供了机会，汇集来自不同学科的研究人员和利益相关者，包括电气和计算机工程，生物工程，分子生物学，神经科学和数据科学，以开发一种面向系统的方法。此外，该项目还通过妇女工程研究奖学金和第一年创新和研究经验（FIRE）等方案，促进历来在工程领域代表性不足的妇女和本科生的参与。多学科的工程方法是必不可少的，以建立一个体外发现平台，能够直接监测化学转导模式沿着肠神经元轴。在任务1中，电化学传感器将直接制造在多孔细胞培养基质上，允许直接进入体外模型肠上皮的细胞和分子机制。细胞层的阻抗监测将检测随时间的物理变化（例如，屏障完整性）。电位监测将检测由于细菌代谢物刺激而从肠细胞释放的实时血清素。在任务2中，将通过将该细胞释放的5-羟色胺暴露于具有连接和受神经支配的后肠的分离的离体小龙虾神经索来研究肠5-羟色胺信号传导的神经效应。将在后肠痉挛期间记录双向连接中枢和肠神经系统的运动和感觉神经元中的神经行为激活模式。机器学习方法将识别关键变量，以量化离散的5-羟色胺释放和神经元激活模式。在任务3中，肠道上皮的粘膜层将被特定的肠道微生物定殖，以评估细菌对屏障完整性、5-羟色胺释放模式和导致的神经肌肉激活的影响。通过机器学习进行分类将量化不同微生物组合对时间依赖性5-羟色胺释放曲线和下游效应的整体和协同效应。这项工作有许多新颖的方面。首先，这是一个新的平台，实现了广泛的集成细胞界面传感器，可直接访问实时细胞和分子数据。其次，使用这项技术来研究肠道和神经系统之间的相互作用，可以对庞大且相对难以接近的肠-脑转导通路提供前所未有的见解。第三，机器学习分析可以识别有意义的神经元通讯模式，并预测肠道细菌对神经行为的预期影响。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

A Hybrid Biomonitoring System for Gut-Neuron Communication

• DOI: 10.1109/jmems.2020.3000392
• 发表时间: 2020-06
• 期刊: Journal of Microelectromechanical Systems
• 影响因子: 2.7
• 作者: A. Chapin;Jin-Gu Han;Ta Ho;J. Herberholz;R. Ghodssi

A portable electrochemical sensing platform for serotonin detection based on surface-modified carbon fiber microelectrodes

• DOI: 10.1039/d2ay01627c
• 发表时间: 2023-01-26
• 期刊: ANALYTICAL METHODS
• 影响因子: 3.1
• 作者: Han，Jinjing;Stine，Justin M.;Ghodssi，Reza
• 通讯作者: Ghodssi，Reza

A Coculture Based Tyrosine-Tyrosinase Electrochemical Gene Circuit for Connecting Cellular Communication with Electronic Networks

基于共培养的酪氨酸-酪氨酸酶电化学基因电路，用于连接细胞通信与电子网络

• DOI: 10.1021/acssynbio.9b00469
• 发表时间: 2020
• 期刊: ACS Synthetic Biology
• 影响因子: 4.7
• 作者: VanArsdale, Eric;Hörnström, David;Sjöberg, Gustav;Järbur, Ida;Pitzer, Juliana;Payne, Gregory F.;van Maris, Antonius J.;Bentley, William E.
• 通讯作者: Bentley, William E.