Autonomous Optogenetic Inhibition of Epileptic Activity Using a Bioluminescent Li

描述（由申请人提供）： 大约20-40%的癫痫患者有难治性癫痫发作，对药物治疗无反应。因此，有必要为这一大群癫痫相关残疾风险较高的人开发替代治疗方法。光遗传学提供了一种以细胞类型特异性方式激发和/或抑制神经活动的强大方法，具有巨大的潜力， 限制或消除作为癫痫发作基础的病理性神经活动的扩散。尽管有这种治疗潜力，但光遗传学的使用面临许多技术挑战，限制了其在临床环境中的实用性和可转化性。在这个建议中，我们提出了一个高度创新的解决方案，这些问题相结合的使用光遗传学与生物发光报告。钙敏感性神经元酶是一种生物发光报告基因，已成功地用于活体神经活动的成像。这些酶通过发射与各种抑制性光敏离子通道的吸收光谱相容的光来响应去极化相关的钙内流。我们建议使用这些报告者来创建我们所说的自主生物控制器，能够驱动光遗传学反馈来控制病理活动。在我们提出的模型中，钙敏感性神经元酶将以光的形式报告神经活动，抑制性视蛋白将被发射的光激活，并且当细胞被视蛋白超极化时，活动的传播将被抑制。癫痫活动的这种自主生物反馈通过消除对外部光源的需要并提供自主闭环反馈控制的手段，为光遗传学的技术局限性提供了一种新的解决方案。我们认为这项研究具有高度创新性，有可能加速光遗传学作为开发癫痫新疗法的工具。

DESCRIPTION (provided by applicant): Approximately 20-40% of patients with epilepsy have refractory seizures unresponsive to pharmacotherapy. There is therefore a need to develop alternative treatments for this large population of people who are at higher risk of developing epilepsy-related disabilities. Optogenetics - which provides a powerful approach to excite and/or inhibit neural activity in a cell-type specific manner - possesses great potential to limit or abolish the spread of the pathological neural activity underlying epileptic seizures. Despite this therapeutic potential, the use of optogenetics faces many technical challenges that limit its usefulness and translatability to the clinical setting. In this proposal, we present a hihly innovative solution to these problems by combining the use of optogenetics with bioluminescent reporters. Calcium-sensitive luciferases are a kind of bioluminescent reporter that has been successfully used for imaging neural activity in vivo. These luciferases respond to depolarization-associated calcium influx by emitting light that is compatible with the absorption spectrum of various inhibitory light-sensitive ion channels. We propose to use these reporters to create what we term an autonomous biologic controller capable of driving optogenetic feedback to control pathological activity. In our proposed model, calcium-sensitive luciferases would report neural activity in the form of light, inhibitory opsins would be activated by the emitted liht, and propagation of activity would be inhibited as the cell is hyperpolarized by the opsin. This autonomous, biological feedback of epileptic activity provides a novel solution to the technical limitations of optogenetics by eliminating the need for an external light source and offering a means for autonomous closed-loop feedback control. We suggest that this research is highly innovative and has the potential of accelerating the use of optogenetics as a tool to develop new therapies for epilepsy.