Remote Electromagnetic Control of Neural Activity for Treatment of Parkinson's Disease

项目摘要 在这种协作和跨学科的应用中，我们建议进一步开发一种新的非侵入性 适用于临床前概念验证研究的细胞调节方法(NICR)。这项技术 潜在地可以用于治疗神经系统疾病，并提供一种侵入性较小的替代深部大脑的方法 刺激(DBS)或光遗传学。因此，我们建议改进技术并开发一个原型设备来 测试NICR对帕金森病(PD)小鼠症状的治疗作用。细胞活动是 由两种成分控制；与铁结合的铁蛋白自发形成5纳米铁 纳米颗粒和TRPV1，一种温度和机械敏感的通道。通过将铁蛋白拴在TRPV1上，1 可以用射频(RF)(加热或诱导铁蛋白的机械运动)或磁铁来控制通道 (这会引起运动)。该方法已被证明能够在体外和体内控制神经活动。 活体，后者通过增加神经放电来实现。此外，我们还在TRPV1中引入了一种突变，它可以将 它进入氯离子通道，而突变通道的使用使抑制神经活动成为可能 电磁波(例如，射频)。因为这个系统是基因编码的，所以人们可以调节 通过重组腺病毒将系统的两个蛋白质组分导入的细胞-- 相关病毒(AAV)株。AAV已被用于包括帕金森病患者在内的许多人体研究。 因此，NICR可以提供一种比植入电极(DBS)或植入的光设备侵入性更小的替代方案 (光遗传学)用于调节神经活动(脑深部刺激)，也可同时用于 控制神经电路中的几个不同节点。 在这个应用中，我们提出了一套治疗帕金森病的临床前概念验证研究 包括：1)改进技术以提高其效率并创造合适的AAV毒株以 改善帕金森病的症状。我们还建议通过使用通道来提高系统的灵敏度 可以在较低的场强下进行选通，并通过识别铁蛋白的变异体来增强对 电磁场；2)开发一种原型装置，它将产生 适当的强度，目的是使该方法能够在常规实验室环境中使用，并最终作为 便携式/可穿戴设备；3)测试改进的方法和适当的仪器设备的能力 减轻帕金森病小鼠的症状；以及4)建立依赖于cre表达的敲击小鼠 构建评估长期TRPV1和铁蛋白表达的安全性。这项技术的验证 还可以用于治疗神经系统内外的其他疾病，以 增加或减少细胞活动或调节蛋白质的产生。最后，NICR的进一步发展 可以通过简单的交配来非侵入性地激活或抑制细胞，从而影响基础研究 转基因小鼠，并将它们暴露在射频或磁场中。

Project Summary In this collaborative and interdisciplinary application, we propose to develop further a novel non-invasive method for cell regulation (NICR) that is suitable for preclinical proof of concept studies. This technology potentially could be used to treat neurologic diseases and provide a less invasive alternative to deep brain stimulation (DBS) or optogenetics. We thus propose to refine the technology and develop a prototype device to test the use of NICR for the treatment of symptoms of Parkinson's Disease (PD) in mice. Cell activity is controlled by two components; the iron binding ferritin protein that spontaneously forms 5 nm iron nanoparticles and TRPV1, a temperature and mechano-sensitive channel. By tethering ferritin to TRPV1, one can gate the channel with radiofrequency (RF) (which heat or induce mechanical motion of ferritin) or a magnet (which induces motion). The method has been shown to be capable of controlling neural activity in vitro and in vivo, the latter by increasing neural firing. In addition, we have introduced a mutation into TRPV1 that converts it into a chloride channel, and the use of the mutant channel makes it possible to inhibit neural activity using electromagnetic waves (e.g., RF). Because the system is genetically encoded, one can regulate the activity of cells into which the two protein components of the system have been delivered by recombinant Adeno- Associated Virus (AAV) strains. AAV has been used in numerous human studies including patients with PD. Thus NICR could provide a less invasive alternative to implanted electrodes (DBS) or implanted light devices (optogenetics) for the modulation of neural activity (deep brain stimulation) and also be used to simultaneously control several different nodes in a neural circuit. In this application, we propose a set of preclinical proof-of-concept studies for the treatment of PD including: 1) refinement of the technology to improve its efficiency and to create suitable AAV strains to ameliorate the symptoms of PD. We also propose to increase the sensitivity of the system by using channels that can be gated with lower field strength and by identifying variants of ferritin with enhanced sensitivity to an electromagnetic field; 2) development of a prototype device that would create local electromagnetic fields of suitable strength with the aim of enabling the use of the method in routine laboratory settings and ultimately as a portable/wearable device; 3) testing the ability of the improved method and suitable instrumentation to alleviate the symptoms of PD in mice; and 4) creating knockin mice with cre dependent expression of the constructs to assess the safety of long term TRPV1 and ferritin expression. The validation of this technology could also lead to its use for the treatment of other diseases at sites within and outside the nervous system to either increase or decrease cell activity or regulate protein production. Finally, the further development of NICR could impact basic research by allowing the non-invasive activation or inhibition of cells by simply mating genetically modified mice and exposing them to RF or magnetic fields.