Bioluminescent optogenetics to promote axon regeneration

生物发光光遗传学促进轴突再生

• 批准号: 9979122
• 负责人: Arthur W. English
• 金额: $ 42.9万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2022-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9979122
• 关键词: Acute; Address; American; Anatomy; Axon; Biological Assay; Biological Models; Bioluminescence; Cells; Clinical; Development; Dose; Effectiveness; Electric Stimulation; Electrophysiology (science); Environment; Exercise; Exposure to; Goals; Growth; Health Care Costs; High Prevalence; Human; Individual; Injury; Investigational Therapies; Ion Channel; Light; Luciferases; Moderate Exercise; Motor; Mus; Natural regeneration; Nerve; Neuronal Injury; Neurons; Operative Surgical Procedures; Opsin; Outcome Measure; Pain; Pathway interactions; Patients; Peripheral; Peripheral Nerves; Peripheral nerve injury; Persons; Pharmacodynamics; Property; Public Health; Quality of life; Recovery; Recovery of Function; Research; Resolution; Route; Sensory; Technology; Time; Transgenic Mice; Translations; Treatment Effectiveness; adeno-associated viral vector; axon injury; axon regeneration; base; clinical translation; clinically relevant; coelenterazine; comorbidity; effectiveness evaluation; functional outcomes; gene therapy; improved; in vivo bioluminescence imaging; injured; injury and repair; mouse model; nerve injury; neuronal excitability; new technology; novel strategies; novel therapeutics; optimal treatments; optogenetics; pain behavior; prevent; reinnervation; repaired; severe injury; standard care; success; virtual

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.

在大量美国人中发现的外周神经损伤（PNIS）中恢复不良， 是一个重大的公共卫生问题。适度运动和短暂的电刺激具有 成为PNI最有效的实验治疗方法，但将其适应患者 证明使用很困难。生物发光光学遗传学（BL-OG）使用兴奋性发光素， 光感应离子通道与发光荧光素酶融合。当暴露于同类时 底物（Coelenterazine，CTZ），生物发光是由荧光素酶部分和生物发光产生的 附着的Opsin已激活。如果诱导受伤的神经元表达兴奋性发光素，则 它们将被这种生物发光激活，刺激其轴突的再生。在 这个项目我们将使用BL-OG以人类可能使用的方式治疗神经损伤 患者。现在将解决该策略的两个关键方面：调查 CTZ给药以激发引起兴奋性神经元的有效性 Luminopsin，在神经损伤后使用基因疗法，而不会加剧通常的疼痛 伴随PNI；以及使用BL-OG对受伤神经元的激发是否会增强 当神经修复延迟以进行有效的基因治疗时，轴突再生。这 BL-OG期间产生的生物发光将被利用以评估药效学 CTZ在临床相关的外周神经损伤的小鼠模型中。有效性 将使用CTZ对轴突再生进行治疗，将在良好的外围神经损伤的小鼠模型中使用电生理和神经 - 动物学结局测量方法进行研究。我们 预计这项探索性/发展研究的结果将为明确 解决了促进使用令人兴奋的新的障碍的根本障碍 技术。

项目成果

Bioluminescent Optogenetics: A Novel Experimental Therapy to Promote Axon Regeneration after Peripheral Nerve Injury.

• DOI: 10.3390/ijms22137217
• 发表时间: 2021-07-05
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: English AW;Berglund K;Carrasco D;Goebel K;Gross RE;Isaacson R;Mistretta OC;Wynans C
• 通讯作者: Wynans C