Developing a microphysiological system of a humanized Gut-Brain-Axis for age-associated transmissible neuropathologies

项目摘要 该项目的重点是设计一种新的神经微生理系统(MPS)，以满足青少年和 老年脑肠轴(GBA)与健康和疾病GBA由迷走神经所在的神经回路组成 将位于肠道的肠神经系统(ENS)神经元与中枢神经系统(CNS)神经元连接， 从而允许双向通信。因此需要简化的神经通路模型 肠道到大脑，在健康和疾病中的潜在作用机制还没有被很好地理解。这个 迷走神经，主要是位于结状神经节的感觉神经元，负责传递饮食线索和中枢神经系统 肠道细菌的功能，并与神经传递性神经退行性疾病有关，如 帕金森氏病(PD)。因此，捕捉人类大脑-肠轴(GBA)的关键组件的MPS， 包括初级中央后脑神经元、迷走神经和肠神经元，将是一种有价值的工具 了解和操纵年轻和老年组织中的通路功能，并促进发现和 通过神经调节进行治疗。这里的方法描述了激光制造、切割和组装的验证 议员们朝着人性化的GBA迈进。我们的多学科团队共同努力(1)建立了一个原型 GBA-MPS，(2)验证组装的MPS内的3D神经室，(3)展示原始人 神经球来源，以及(4)建立肠道支配结节神经元的转基因模型。两个目标将是 被追捕。AIM 1将设计一种MPS，通过现场电生理和 青年和老年人群神经功能的成像、时间进程和终点表征 与静态和单一培养相比，确保适当的命运和表型。AIM 2将验证以下各项的实用性 通过肠道神经元中的预形成纤维(PFF)建立疾病模型PD表型的平台 年轻和老年MPS，并利用视蛋白表达转基因结节神经元的光刺激，电路 Lynchpin，评估神经调节作为一种方法来减缓帕金森病从肠道扩散到脑室。 首个模拟人类GBA的MPS的成功完成将加速对GBA的机理研究 健康和疾病中的脑-肠道沟通，通过启用分析来促进治疗靶点的发现 神经生物学和神经退行性疾病的一个可访问和仪器化的MPS平台。

Project Summary This project is focused on engineering a novel innervated microphysiological system (MPS) of the young and aged brain-gut axis (GBA) in health and disease. The GBA is comprised of neural circuits where the vagus nerve connects enteric nervous system (ENS) neurons that reside in the gut to central nervous system (CNS) neurons, thus allowing for bi-directional communication. There is a need for simplified models of the neural pathways from gut-to-brain where the underlying mechanisms of action in health and disease are not well understood. The vagus nerve, predominantly sensory neurons located in the Nodose Ganglia, transduces dietary cues and CNS function by gut bacteria and has been implicated in neurotransmittable neurodegenerative diseases such as Parkinson’s Disease (PD). Thus, an MPS that captures key components of the human brain-gut axis (GBA), including primary central hind brain neurons, vagal neurons, and enteric neurons, would be a valuable tool for understanding and manipulating pathway function in young and aged tissue and for advancing discovery and therapies via neuromodulation. The approach here describes validation of a laser-fabricated, cut & assembled MPS towards a humanized GBA. Our multidisciplinary team has worked together to (1) establish a prototype GBA-MPS, (2) validate 3D neural compartments within assembled MPS, (3) demonstrate primary human neurosphere sourcing, and (4) create a transgenic model of gut innervating nodose neurons. Two aims will be pursued. Aim 1 will engineer an MPS with real-time sensing of GBA function via live electrophysiology and imaging, followed by time-course and end point characterization of neural function in young and aged populations compared to static and monocultures to ensure appropriate fate and phenotype. Aim 2 will validate the utility of the platform to create a disease model PD phenotype by pre formed fibrils (PFFs) in the enteric neurons of the young and aged MPS, and harness photostimulation of opsin expressing transgeneic nodose neurons, the circuit lynchpin, to evalutate neuromodulation as a method to slow PD spread from the gut to the brain compartment. Successful completion of the first ever MPS that simulates human GBA will accelerate the mechanistic study of brain-gut communication in health and disease and advance therapeutic target discovery by enabling analysis of neural biology and neurodegenerative disorders an accessible and instrumented MPS platform.