靶向酸敏感离子通道调控新机制的系统药理学研究

Acid-sensing ion channel (ASIC) is a type of cation channels activated by extracellular acidification and is ubiquitously expressed throughout the body, which probably constitutes a potential drug target for the treatment of various diseases. We are long engaging in research on the function and regulation of ASIC, and have pioneered the identification of ASIC action in acidotoxicity during the pathological process in the disease models of stoke and chronic pain. In addition, we proofed the concept of the nonproton ligand sensor in ASIC, and screened a number of novel exogenous and endogenous nonproton ligands for the channel. Furthermore, we identified a novel metabolic signal pathway mediated by ASIC, which expands the non-conducting mechanism for the channel complexes. Based on these progresses, here we propose to deepen the functional validation studies on drugablity of ASIC-mediated whole signaling cascades in stroke and chronic pain, by taking advantages of conventional and conditional ASIC knockout mice, introducing CRISPR/Cas9 gene editing animals with loss-of-function or gain-of-function mutants on particular aspect of ASIC signaling, developing chemical biology tools, in addition to using a variety of physiological assays including electrophysiology, cell culture, disease models, and behavioral analysis. We expect to develop novel screening models of pharmacological activity on ASIC, discover several lead compounds with novel molecular structures that are effective on the ASIC activity, characterize the pharmacological activity of these compounds at the molecular and cellular as well as the whole animal levels, in order to elucidate the underlying mechanisms and comprehensively verify the causal relationship with the ASIC as the drug target. The final aim is to provide further theoretical basis and practical guidance for the clinical application of ASIC-targeted therapy.

酸敏感离子通道（ASIC）是一类被胞外酸化激活的阳离子通道，在体内分布广泛，可能成为多种疾病治疗的潜在药物靶标。基于脑卒中和慢性痛等疾病模型，申请人所在项目组揭示了ASIC介导酸毒性神经损伤的分子机制；建立和发展了ASIC非质子配体感受域的新概念，筛选出多个非质子新配体；鉴定了ASIC代谢性信号转导新途径，拓展了ASIC的非离子通道作用机制。本项目旨在前期工作的基础上，借助于基因敲除小鼠，引入CRISPR-Cas9基因编辑的离子通道特定作用机制选择性缺失或获得的动物模型，同时发展化学生物学手段，综合运用电生理学、细胞培养、疾病模型和行为学分析等功能研究方法，基于ASIC信号转导的完整过程开展成药性的功能确证研究，通过发展药理活性筛选模型，发现结构新颖的先导化合物，在分子、细胞和整体动物水平研究先导化合物的药理活性并阐明分子机制，为开展靶向ASIC疾病治疗的临床应用提供理论基础和实践指导。

酸敏感离子通道（ASICs）是一类被胞外酸化激活的阳离子通道，在体内分布广泛，可能成为多种疾病治疗的潜在药物靶标。基于其在脑卒中和慢性痛等神经系统疾病中的重要作用，本项目围绕ASIC靶标的结构调控与病理功能这一核心问题，按计划借助于多种基因修饰模型及高通量筛选手段，深入解析了ASIC1a代谢性信号机制在缺血性神经损伤中的核心作用，描绘了ASICs的非离子通道作用途径，阐释了ASIC1a介导的中枢可塑性在慢性痛及其他神经精神疾病中的贡献，鉴定了ASIC3在神经免疫互作及相关感觉异常过程中的作用。通过发展药理活性筛选模型，项目系统性确证了药理活性与ASICs介导特定信号转导途径相关性，为最终开展靶向ASICs疾病治疗的临床应用提供理论基础和实践指导。项目在《Nature Communications》、《Science Advances》、《PNAS》、《Redox Biology》等国际学术期刊发表论文13篇，获中国授权专利3项，4项申请专利等待获批；筛选到靶向ASICs的数个全新小分子及单克隆抗体，在动物模型上确证了ASICs在神经保护和镇痛等方面的药理学可靶性。培养博士研究生毕业8人，博士后出站3人，以联合培养的方式为国内合作单位培养研究生毕业2人。培养项目组成员获各类资助、奖励或人才计划6项；项目组参与组织学术会议7次，接待学术来访交流14场，项目组成员参加国内外学术会议42人次，圆满完成了预期目标。