Bioluminescent optogenetics to promote axon regeneration

在大量美国人中发现的周围神经损伤(PNIS)恢复较差， 是一个重大的公共卫生问题。适度的运动和短暂的电刺激有 成为治疗PNI最有效的实验性治疗方法，但要根据患者的情况进行调整 事实证明，使用起来很困难。生物发光光遗传学(BL-OG)使用兴奋性发光素， 感光离子通道与发光的荧光素酶融合。当接触到同源词时 底物(coelenterazine，CTZ)，生物发光是由荧光素酶部分和 附着的视蛋白被激活。如果损伤的神经元被诱导表达兴奋性发光素， 它们会被这种生物发光激活，刺激轴突的再生。在……里面 在这个项目中，我们将使用BL-OG来治疗神经损伤，这种方式可能会用于人类 病人。现在将讨论该战略的两个关键方面：调查 CTZ对兴奋诱导的神经元表达兴奋性的影响 使用发光素，在神经损伤后使用基因治疗，而不会加剧经常发生的疼痛 以及使用BL-OG对受损神经元的这种兴奋是否会增强 当神经修复延迟时轴突再生，以便进行有效的基因治疗。这个 将利用BL-OG过程中产生的生物发光来评估药效学 CTZ在临床相关的周围神经损伤小鼠模型中的作用。的有效性。 CTZ对轴突再生的治疗将使用电生理学和神经解剖学结果指标在已建立的周围神经损伤小鼠模型中进行研究。我们 预期这一探索性/发展性研究的结果将提供一个明确的 解决推动使用令人振奋的新技术的根本障碍 技术

The poor recovery from peripheral nerve injuries (PNIs), found in a large number of Americans, is a significant public health issue. Moderate exercise and brief electrical stimulation have emerged as the most effective experimental treatments for PNI, but adapting them to patient use has proven difficult. Bioluminescent optogenetics (BL-OG) uses excitatory luminopsins, light-sensing ion channels fused with a light-emitting luciferase. When exposed to a cognate substrate (coelenterazine, CTZ), bioluminescence is generated by the luciferase moiety and the attached opsin is activated. If injured neurons are induced to express an excitatory luminopsin, they would be activated by this bioluminescence, stimulating the regeneration of their axons. In this project we will use BL-OG to treat nerve injuries in a manner that might be used in human patients. Two critical aspects of that strategy will be addressed now: Investigating the effectiveness of CTZ administration to excite neurons induced to express an excitatory luminopsin, using gene therapy after nerve injury, without exacerbating the pain that often accompanies PNI; and whether this excitation of injured neurons using BL-OG will enhance axon regeneration when nerve repair is delayed to allow for effective gene therapy. The bioluminescence produced during BL-OG will be exploited to evaluate the pharmacodynamics of CTZ in a clinically relevant mouse model of peripheral nerve injury. The effectiveness of treatment with CTZ on axon regeneration will be studied using electrophysiological and neuro-anatomical outcome measures in a well-established mouse model of peripheral nerve injury. We anticipate that the results of this exploratory/developmental research will provide a clear resolution to the fundamental obstacle to advancement of the use of an exciting new technology.