Topology and Mechanism of Broadband and Fast Multi-Characteristic Opsin for Neuromodulation

IF 2.6 4区 医学 Q2 PHARMACOLOGY & PHARMACY
Samarendra Mohanty, Pratyajit Mohapatra, Amir Singh, William Marks, Subrata Batabyal, Michael Carlson, Najam Sharif, Sanghoon Kim
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Abstract

Optogenetics has enabled targeted manipulation of neural circuits, offering insights into the intricacies of brain function and paving the way for potential therapeutic applications in neurological disorders. Multi-characteristic Opsin (MCO) is an optogenetics therapy candidate that is undergoing multiple clinical trials, including randomized control trials. However, its cellular topography and mechanism of action have remained unknown. Here, we reveal the biophysical characteristics of the MCO molecule, including its unique structure and function, using a combination of predictive protein modeling, western blotting-based protease protection assay, and electrophysiology. MCO’s first extracellular subunit forms a cation-channel pore, while the second subunit is partially embedded in the membrane, extending to the third intracellular cytoplasmic subunit. Functional studies using whole-cell recordings showed that MCO is activated by broadband visible light with fast on-off kinetics with high photosensitivity, and large dynamic range across the visible spectrum, enabling best-in-class light-activation of MCO-expressing cells. Further, multi-electrode array recording confirmed multi-color light-activation of MCO-expressing neurons in the retina. The unique topological and functional activation features of fused MCO protein represent novel findings linking the in-vitro and in-vivo efficacy of this opsin, which is particularly important for neuromodulation leading to vision restoration in retinal degenerative diseases and other neurological therapies.

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宽带和快速多特征视蛋白的神经调节拓扑结构和机制
光遗传学使神经回路的定向操作成为可能,为大脑功能的复杂性提供了见解,并为神经系统疾病的潜在治疗应用铺平了道路。多特征视蛋白(MCO)是一种光遗传学治疗候选药物,目前正在进行包括随机对照试验在内的多项临床试验。然而,其细胞形态和作用机制仍不清楚。在这里,我们揭示了MCO分子的生物物理特性,包括其独特的结构和功能,结合预测蛋白质模型,基于western blotting的蛋白酶保护实验和电生理学。MCO的第一个胞外亚基形成阳离子通道孔,而第二个亚基部分嵌入膜中,延伸到第三个胞内细胞质亚基。利用全细胞记录的功能研究表明,MCO可以被宽带可见光激活,具有高光敏性和快速的开关动力学,以及在可见光谱上的大动态范围,使表达MCO的细胞具有同类最佳的光激活能力。此外,多电极阵列记录证实了视网膜中表达mco的神经元的多色光激活。融合MCO蛋白独特的拓扑结构和功能激活特征代表了这种视蛋白在体外和体内功效的新发现,这对于视网膜退行性疾病和其他神经治疗中导致视力恢复的神经调节尤为重要。
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来源期刊
Advanced Therapeutics
Advanced Therapeutics Pharmacology, Toxicology and Pharmaceutics-Pharmaceutical Science
CiteScore
7.10
自引率
2.20%
发文量
130
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