Optogenetic inhibition of motor neuron and muscle activity in vivo

体内运动神经元和肌肉活动的光遗传学抑制

基本信息

批准号：
8661322
负责人：
SCOTT L DELP
金额：
$ 34.2万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8661322
关键词：
Action Potentials Activities of Daily Living Afferent Neurons Animals Axon Botulinum Toxins Cerebral Palsy Chloride Ion Chlorides Chronic Clinical Development Devices Disease Dorsal Electrodes Foundations Future Genes Genetic Goals Halorhodopsins Hindlimb Human Implant Injection of therapeutic agent Intramuscular Ion Channel Laboratories Lasers Lead Life Light Mediating Membrane Potentials Methods Motor Neurons Movement Mus Muscle Muscle Contraction Nerve Nervous System Trauma Neurons Neurotoxins Operative Surgical Procedures Opsin Optics Oral Paralysed Performance Peripheral Peripheral Nerves Peripheral Nervous System Pharmaceutical Preparations Physiologic pulse Property Pump Reporting Rhizotomy procedure Rodent Smooth Muscle Speech Spinal cord injury Stroke System Techniques Technology Testing Time Transgenic Mice Transgenic Organisms Translating Viral Viral Vector Virus Work base design gene therapy implantation improved in vivo nervous system disorder neuronal cell body neuroregulation new technology novel novel strategies optogenetics prevent relating to nervous system response retrograde transport sciatic nerve trafficking uptake

项目摘要

DESCRIPTION (provided by applicant): Neurological disorders and injuries frequently result in spasticity and involuntary muscle contractions that interfere with speech, movement and activities of daily living. Current approaches to reduce spasticity, such as selective dorsal rhizotomy, oral medications, or injections of botulinum toxin, have important limitations. There is currently no treatment for spasticity that provides targeted, tunable and rapidly reversible inhibition of unwanted muscle activity. This proposal describes a novel strategy to achieve precise, tunable, and reversible inhibition of motor neuron and muscle activity in vivo. Our approach is made possible by the recent discovery of optogenetics, a technique that enables the use of light-sensitive ion channels to facilitate optical excitation or inhibition of mammalian neurons. Our laboratory has recently reported the first use of optogenetics to excite sciatic nerve motor neurons in transgenic mice. However, it is unknown if a similar approach can be used to inhibit motor neuron activity using halorhodopsin (NpHR), a light sensitive ion channel that hyperpolarizes axons in response to light and impedes action potential propagation along a nerve. Our first aim is to apply light to the sciatic nerve of transgenic mice that express NpHR in their motor neuron axons. We will ask whether the application of light can block action potentials induced by electric nerve stimulation in a tunable and reversible manner and thereby prevent muscle contraction. Our preliminary results in anesthetized animals indicate that such inhibition is not only possible, but is repeatable and robust. After we characterize the light delivery properties required for effective inhibition, we will build an implantable light delivery device an use it to achieve motor neuron inhibition in freely moving transgenic mice. Finally, as a necessary prelude to future clinical use of this technology, we will translate this work to non-transgenic animals. For this purpose we will use state of the art gene therapy techniques to selectively deliver the NpHR gene to targeted motor neuron pools in a manner that is readily translated to humans. This project will lay the foundation for optogenetic modulation of activity i motor neurons and other peripheral nerves. Successful demonstration of optical inhibition will provide a proof-of-principle for a novel treatment for spasticity. Various components of this project, including techniques for opsin delivery to target nerves and the development of an implantable light delivery device, are needed to enable optogenetics-based treatments for a variety of other disorders.

描述（由申请人提供）：神经系统疾病和伤害经常导致痉挛和非自愿性肌肉收缩干扰言语，运动和日常生活的活动。当前减少痉挛的方法，例如选择性背根茎，口服药物或肉毒杆菌毒素的注射具有重要局限性。有目前尚无痉挛的治疗方法，可提供对不良肌肉活动的靶向，可调节和快速可逆的抑制作用。该提案描述了一种新的策略，以实现对体内运动神经元和肌肉活动的精确，可调和可逆的抑制作用。通过最近发现的光遗传学使我们的方法成为可能，该技术使光敏离子通道可以促进光激发或抑制哺乳动物神经元。我们的实验室最近报道了转基因小鼠中首次使用光遗传学来激发坐骨神经运动神经元。但是，尚不清楚使用类似的方法使用甲状腺素（NPHR）抑制运动神经元活性，这是一种光敏的离子通道，可响应光线响应光并阻碍沿神经沿神经的动作电位传播。我们的第一个目的是将光施加到表达NPHR的转基因小鼠的坐骨神经他们的运动神经元轴突。我们将询问光的应用是否可以以可调和可逆的方式阻止电神经刺激引起的动作电位，从而防止肌肉收缩。我们在麻醉动物中的初步结果表明，这种抑制不仅是可能的，而且是可重复且健壮的。在表征有效抑制所需的光递送特性之后，我们将建立一个可植入的光递送装置，以实现自由移动的转基因小鼠的运动神经元抑制。最后，作为该技术未来临床使用的必要前奏，我们将将这项工作转化为非转基因动物。为此，我们将使用ART基因治疗技术的状态将NPHR基因选择性地传递给靶向运动神经元池，以容易转化为人类。该项目将奠定活性I运动神经元和其他周围神经的光学调节基础。成功证明光学抑制作用将为新颖的痉挛治疗提供原则证明。该项目的各种组成部分，包括针对靶神经的Opsin递送技术以及开发可植入的轻型递送装置，需要为其他各种疾病提供基于光遗传学的治疗方法。