Ion Channel Screening Using Photoswitches

使用光电开关进行离子通道筛选

• 批准号: 8646676
• 负责人: Andrew Blatz
• 金额: $ 19.33万
• 依托单位: PHOTOSWITCH BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8646676
• 关键词: Action Potentials; Affect; Arrhythmia; Award; Behavior; Biological Assay; Budgets; Calcium Channel; Cell Adhesion; Cell Line; Cell membrane; Cells; Cellular Membrane; Charge; Computer software; Coupled; Cystic Fibrosis; Data; Detection; Development; Disease; Dose; Drug Targeting; Dyes; Economics; Electrodes; Electrophysiology (science); Engineering; Equipment Design; Event; Fluorescence Resonance Energy Transfer; Funding; Goals; Heart; Hybrids; Hypertension; Industry; Inhibitory Concentration 50; Ion Channel; Light; Lighting; Liquid substance; Measurement; Measures; Mediating; Membrane Potentials; Membrane Proteins; Methods; Movement; Muscle; Muscular Dystrophies; Nervous system structure; Neurons; Noise; Optics; Pathology; Pathway interactions; Performance; Pharmaceutical Preparations; Phase; Physiological; Price; Procedures; Proteins; Reagent; Reporting; Scheme; Signal Transduction; Small Business Innovation Research Grant; Sodium Channel; Source; System; Techniques; Technology; Testing; Therapeutic Intervention; Whole-Cell Recordings; Work; assay development; base; commercialization; design; drug discovery; electric field; extracellular; falls; instrument; instrumentation; interest; nanomachine; novel; operation; programs; prototype; public health relevance; response; screening; software development; stable cell line; success; voltage

Ion channels are membrane proteins that allow the selective movement of charged molecules through cellular membranes. Many disease states, such as cystic fibrosis, muscular dystrophies, hypertension, and cardiac arrhythmias are caused by ion channel pathologies. Ion channels have been identified by the drug discovery industry as excellent targets for therapeutic intervention but because of the technical hurdles involved in developing quality, high- throughput functional ion channel assays, the development of "ion channel drugs" has been disappointing. Photoswitch Biosciences has developed technology, which allows attaching small nanomachines to ion channels enabling rapid and reversible control of these proteins by light. These "photoswitched" ion channels allow us to non- invasively control cell membrane potential and, hence, the activity of voltage-dependent ion channels. The Specific Aim for the Phase I component of this fast track SBIR project are: (1) Demonstrate the feasibility of using extracellular electric field recording as a membrane potential readout. Our original plan was to use voltage- sensitive dyes for this application, but recent results from our lab suggests that extracellular recording of electrical field potentials may offer a variety of benefits, including instrument design and assay simplicity. The Phase II goals are to build three fully functional prototype screening systems consisting of instruments, engineered cell lines, reagents, and software. The use of the hybrid method of extracellular field potential recording technology coupled with photoswitch technology for ion channel assays has several advantages over existing techniques: (1) Elimination of liquid additions for channel activation, (2) Highly scalable for high-throughput ion channel drug discovery, (3) Price per data point is at least 100X less than automated electrophysiology, (4) Channel activation is almost 100X faster than KCl addition. The ability to rapidly depolarize and repolarize cells in a high-throughput format is especially important for the discovery of "use-dependent" compounds that achieve target selectivity via affecting only the channels that are active. This breakthrough technology will finally allow the full exploitation of ion channels as drug targets.

离子通道是膜蛋白，允许带电分子通过细胞选择性移动 膜。许多疾病状态，如囊性纤维化、肌营养不良、高血压和心律失常 是由离子通道病理引起的。离子通道已被药物发现行业认定为优秀的 治疗干预的目标，但由于开发高质量、高水平的技术障碍 尽管功能性离子通道检测的吞吐量较高，但“离子通道药物”的开发却令人失望。 Photoswitch Biosciences 开发了一种技术，可以将小型纳米机器连接到离子通道 能够通过光快速且可逆地控制这些蛋白质。这些“光开关”离子通道使我们能够非 侵入性地控制细胞膜电位，从而控制电压依赖性离子通道的活性。 该快速轨道 SBIR 项目第一阶段的具体目标是： (1) 论证 使用细胞外电场记录作为膜电位读数。我们最初的计划是使用电压- 对于这种应用敏感染料，但我们实验室的最新结果表明，细胞外的电记录 场电位可以带来多种好处，包括仪器设计和测定的简单性。第二阶段目标 将建立三个功能齐全的原型筛选系统，包括仪器、工程细胞系、 试剂和软件。利用混合法耦合细胞外场电位记录技术 与现有技术相比，采用光电开关技术进行离子通道测定具有以下几个优点： (1) 消除 用于通道激活的液体添加，(2) 高度可扩展，用于高通量离子通道药物发现，(3) 每个数据点的价格至少比自动电生理学低 100 倍，(4) 通道激活几乎是 100 倍 比 KCl 添加速度更快。以高通量形式快速去极化和复极化细胞的能力是 对于发现“依赖于用途”的化合物尤其重要，这些化合物仅通过影响即可实现目标选择性 活跃的频道。这项突破性技术最终将允许离子通道的充分利用 药物靶点。