Ion Channel Screening Using Photoswitches

使用光电开关进行离子通道筛选

• 批准号: 8887728
• 负责人: Andrew Blatz
• 金额: $ 59.62万
• 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 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8887728
• 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; Health; 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; response; screening; software development; stable cell line; success; voltage

DESCRIPTION (provided by applicant): 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 offera 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.

描述（由申请人提供）：离子通道是允许带电分子选择性移动通过细胞膜的膜蛋白。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鉴定为治疗干预的优良靶标，但由于涉及开发高质量、高通量功能性离子通道测定的技术障碍，“离子通道药物”的开发一直令人失望. 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上，从而通过光快速可逆地控制这些蛋白质。这些“光开关”离子通道允许我们非侵入性地控制细胞膜电位，并因此控制电压依赖性离子通道的活性。本快速SBIR项目第一阶段的具体目标是：（1）证明使用细胞外电场记录作为膜电位的可行性 读数。我们最初的计划是使用电压敏感染料用于这一应用，但我们实验室最近的结果表明，电场电位的细胞外记录可能提供各种好处，包括仪器设计和测定简单性。第二阶段的目标是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。使用细胞外场电位记录技术结合光开关技术的混合方法进行离子通道测定与现有技术相比具有几个优点：（1）消除了用于通道激活的液体添加，（2）对于高通量离子通道药物发现高度可扩展，（3）每个数据点的价格比自动电生理学低至少100倍，（4）通道激活比KCl添加快几乎100倍。以高通量形式快速去活化和再活化细胞的能力对于发现“使用依赖性”化合物特别重要，所述化合物通过仅影响活性通道来实现靶选择性。这项突破性的技术将最终允许充分利用离子通道作为药物靶点。