Autonomous Optogenetic Inhibition of Epileptic Activity Using a Bioluminescent Li

描述(申请人提供)：大约20-40%的癫痫患者有难治性发作，对药物治疗无效。因此，有必要为患癫痫相关残疾的风险较高的这一大群人开发替代疗法。光遗传学-它提供了一种强大的方法来以细胞类型特定的方式兴奋和/或抑制神经活动-具有巨大的潜力 限制或取消癫痫发作背后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有用性和可译性。在这项提案中，我们提出了一种通过结合使用光遗传学和生物发光记者来解决这些问题的高度创新的解决方案。钙敏感的荧光素酶是一种生物发光报告，已成功地用于体内神经活动的成像。这些荧光素酶通过发出与各种抑制性光敏离子通道的吸收光谱相兼容的光来响应去偏振相关的钙内流。我们建议使用这些记者来创造我们所说的能够驱动光遗传反馈来控制病理活动的自主生物控制器。在我们提出的模型中，钙敏感的荧光素酶将以光的形式报告神经活动，抑制性视蛋白将被释放的LiHT激活，并且由于视蛋白的超极化，活动的传播将被抑制。这种癫痫活动的自主生物反馈通过消除对外部光源的需求并提供自主闭环反馈控制的手段，为光遗传学的技术限制提供了一种新的解决方案。我们认为，这项研究具有很高的创新性，有可能加速利用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.