A 3D-Printed Nerve Cuff for 1-Photon Optogenetic Vagal Stimulation

项目总结/摘要 越来越多地考虑使用自主神经系统的外周神经的治疗性干预， 用于调节器官功能以治疗疾病。迷走神经刺激（VNS）已获得临床批准 二十多年来一直用于治疗癫痫发作和抑郁症。它还表现出积极的临床 治疗炎症性疾病如类风湿性关节炎和克罗恩病的结果， 临床前数据表明在心脏病和糖尿病中的治疗益处。胆碱能（副交感神经）通路是 被认为是观察到的治疗效果的核心。“胆碱能抗炎通路”是一种 已建立的信号传导机制，通过该机制，促炎细胞因子的表达和释放减少， 脾脏和其他内脏组织中。因为炎症是许多疾病的驱动因素， 炎性介质是主要关注的。 虽然这些神经通路具有很好的治疗潜力，但目前的设备具有明显的局限性。电极- 在现有VNS治疗中使用的基于装置将电流广泛地施加到神经。这种电气的非特定性质 刺激激活脱靶通路，对患者造成不利影响，并且此外， 足够的工具来研究和理解治疗相关的神经通路。这项研究将开发一种新的 用于光遗传迷走神经刺激的装置，可以消除这些脱靶并发症。利用光遗传学工具， 光敏致动器可以通过遗传靶向和组织特异性病毒递送靶向感兴趣的细胞类型， 从而能够高度特异性地激活和研究神经回路。 目前的研究将开发和验证用于光遗传学研究的3D打印迷走神经袖带系统。新型微型 分辨率立体光刻3D打印能力将被用来生产生物相容性，单组分， 光刺激神经植入物（Aim 1）。这种方法将实现易于制造和易于植入的装置， 可以应用于研究和识别许多疾病状态的神经回路。该器械将在 转基因动物模型（Aim 2），其中胆碱能途径将在以下小鼠模型中被光遗传学刺激： 急性和慢性全身性炎症。促炎介质的调节将在体内测量， 为期沿着用胆碱能-迷走神经心率反应测试系统的功效。据推测 该装置将实现用于慢性光遗传迷走神经刺激的简单使用的系统， 在动物模型中的全身炎症介质。

Project Summary/Abstract Therapeutic interventions using peripheral nerves of the autonomic nervous system are increasingly being considered and applied to modulate organ function for disease treatments. Vagus nerve stimulation (VNS) has been clinically approved for over two decades for its treatment of epileptic seizures and depression. It has also demonstrated positive clinical outcomes in the treatment of inflammatory disorders such as rheumatoid arthritis and Crohn's disease, while clinical and pre-clinical data suggest therapeutic benefit in heart disease and diabetes. Cholinergic (parasympathetic) pathways are believed to be central in the therapeutic effects that are observed. The `cholinergic anti-inflammatory pathway' is an established signaling mechanism by which the expression and release of pro-inflammatory cytokines are reduced in the spleen and in other visceral tissues. Because inflammation is a driver of numerous diseases, the ability to regulate inflammatory mediators is of major interest. While these neural pathways hold promising therapeutic potential, current devices have significant limitations. Electrode- based devices used in existing VNS therapies apply current broadly to the nerve. The non-specific nature of this electrical stimulation activates off-target pathways causing adverse effects to the patient, and furthermore, does not provide an adequate tool to study and understand therapeutically relevant nerve pathways. The proposed research will develop a new device for optogenetic vagus nerve stimulation that can eliminate these off-target complications. With optogenetic tools, light-sensitive actuators can be targeted to cell-types of interest via genetic targeting and tissue-specific viral delivery, thus enabling highly specific activation and investigation of neural circuits. The current study will develop and validate a 3D printed vagus nerve cuff system for optogenetic studies. Novel micro- resolution stereolithographical 3D-printing capabilities will be leveraged to produce a biocompatible, single-component, optical stimulation nerve implant (Aim 1). This approach will enable an easy-to-fabricate and easy-to-implant device that can be applied to study and identify neural circuits across many disease states. The device will be validated in a transgenic animal model (Aim2) in which cholinergic pathways will be optogenetically stimulated in mouse models of acute and chronic systemic inflammation. Modulation of pro-inflammatory mediators will be measured in vivo over a period of two months along with the cholinergic-vagal heart rate response to test efficacy of the system. It is hypothesized that this device will enable a simply-employed system for chronic optogenetic vagus nerve stimulation that will reduce systemic inflammatory mediators in an animal model.