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

项目摘要 在这种合作和跨学科的应用，我们建议进一步开发一种新的非侵入性 细胞调节方法（NICR），适用于临床前概念验证研究。这项技术 有可能用于治疗神经系统疾病，并为深部脑提供侵入性较小的替代方案。 刺激（DBS）或光遗传学。因此，我们建议改进技术并开发原型设备， 测试NICR用于治疗小鼠帕金森病（PD）症状的用途。细胞活性 由两种成分控制;自发形成5 nm铁的铁结合铁蛋白 纳米颗粒和TRPV 1，温度和机械敏感通道。通过将铁蛋白与TRPV 1连接， 可以用射频（RF）（加热或诱导铁蛋白的机械运动）或磁铁门控通道 （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。 体内，后者通过增加神经放电。此外，我们还在TRPV 1中引入了一种突变， 它进入氯离子通道，并且突变通道的使用使得有可能使用 电磁波（例如，RF）。因为这个系统是基因编码的，人们可以调节 所述系统的两种蛋白质组分已经通过重组腺病毒递送到其中的细胞中， 相关病毒（AAV）株。AAV已用于许多人类研究，包括PD患者。 因此，NICR可以提供植入电极（DBS）或植入光装置的侵入性较小的替代方案 （光遗传学）用于调节神经活动（脑深部刺激），也可用于同时 控制神经回路中的几个不同节点 在本申请中，我们提出了一组临床前概念验证研究，用于治疗PD 包括：1）改进技术以提高其效率并产生合适的AAV菌株， 改善PD症状。我们还建议通过使用通道来增加系统的灵敏度 可以用较低的场强门控，并通过识别对铁蛋白的敏感性增强的铁蛋白变体， 电磁场; 2）开发一种原型设备，可以产生局部电磁场， 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）测试改进的方法和合适的仪器的能力， 减轻小鼠中的PD症状;和4）产生具有cre依赖性表达的 构建体以评估长期TRPV 1和铁蛋白表达的安全性。这项技术的验证 也可以导致其用于治疗神经系统内外部位的其他疾病， 增加或减少细胞活性或调节蛋白质产生。最后，对NICR的进一步发展进行了展望。 可以通过简单的交配来非侵入性地激活或抑制细胞， 转基因小鼠并将其暴露于射频或磁场。

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.