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

项目摘要 该项目的重点是工程一种新的神经支配的微生理系统（MPS）的年轻和 老年脑肠轴（GBA）在健康和疾病中的作用。GBA由迷走神经回路组成， 将位于肠道中的肠神经系统（ENS）神经元连接到中枢神经系统（CNS）神经元， 从而允许双向通信。有必要简化模型的神经通路，从 肠道到大脑，其中对健康和疾病的潜在作用机制还没有很好的理解。的 迷走神经，主要是位于结状神经节中的感觉神经元， 肠道细菌的功能，并已牵连在神经传递性神经退行性疾病， 帕金森病（PD）。因此，捕获人脑-肠轴（GBA）的关键组分的MPS， 包括初级中枢后脑神经元，迷走神经元和肠神经元，将是一个有价值的工具， 理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 MPS朝向人源化GBA。我们的多学科团队共同努力（1）建立一个原型 GBA-MPS，（2）确认组装MPS内的3D神经隔室，（3）证明主要人体 神经球来源，和（4）创建肠神经支配结状神经元的转基因模型。两个目标将是 追求。目标1将设计一种MPS，通过实时电生理学实时感知GBA功能， 影像学检查，随后是年轻和老年人群神经功能的时程和终点特征 与静态和单一培养相比，以确保适当的命运和表型。目标2将验证 该平台通过在肠神经元中的预形成原纤维（PFF）来创建疾病模型PD表型， 年轻和老年MPS，并利用光刺激表达视蛋白的转基因结状神经元，电路 lynchpin，以评估神经调节作为减缓PD从肠道扩散到脑室的方法。 成功完成第一个模拟人类GBA的MPS将加速对 健康和疾病中脑-肠通信以及通过启用分析来推进治疗靶点发现 神经生物学和神经退行性疾病的一个可访问的和仪器化的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.