A universal photoswitch system for optical control of neuronal receptors

用于神经元受体光学控制的通用光电开关系统

• 批准号: 7726422
• 负责人: RICHARD H KRAMER
• 金额: $ 30.7万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-22 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7726422
• 关键词: Achievement; Address; Affect; Antibodies; Area; Award; Biochemical; Biology; Brain-Derived Neurotrophic Factor; Cell Surface Proteins; Cell Surface Receptors; Cell physiology; Clinical; Communities; Development; Effectiveness; Electronics; Environment; Family; Funding; G-Protein-Coupled Receptors; Goals; Grant; Growth Factor Receptors; Guidelines; Human Resources; Ion Channel; Leadership; Ligands; Light; Logic; Methodology; Methods; Nature; Neurons; Neurophysiology - biologic function; Neurotrophic Tyrosine Kinase Receptor Type 2; Optics; Outcome; Peptides; Persons; Phase; Photophobia; Population; Probability; Process; Program Reviews; Published Comment; Receptor Activation; Receptor Protein-Tyrosine Kinases; Recording of previous events; Regulation; Research; Research Personnel; Role; System; Techniques; Technology; Testing; TimeLine; Tissues; Training; Translating; United States National Institutes of Health; Work; azobenzene; base; brain-derived growth factor; clinical practice; combinatorial; design; inhibitor/antagonist; innovation; meetings; member; neuropeptide Y; neurotrophic factor; novel; organizational structure; public health relevance; receptor; response; success; tool

DESCRIPTION (provided by applicant): One of the most exciting technical developments in biology in recent years is the emergence of photochemical methods for controlling electrical activity with light. Our goal in this project is to develop a broadly applicable method for controlling other aspects of cellular function by generating tools for conferring light sensitivity on a broad range of cell surface receptors and ion channels. To meet this challenge we will use a modular approach, utilizing a single "Universal Photoswitch" as the key light-sensing component. The photoswitch contains at its core the small isomerizable azobenzene moiety, which shortens and lengthens in response to 380 and 500 nm light, respectively. We will use this photoswitch to indirectly regulate receptor and channel activity, through an "adapter peptide", which contains a "capture domain", which recognizes the short, but not the long configuration of the photoswitch, and a ligand domain, which contains a peptide activator or inhibitor of the targeted cell surface receptor or ion channel. The capture domain is kept constant among all adapter peptides, allowing control by a single Universal Photoswitch, but the nature of ligand domain is tailored to regulate a specific receptor. Several strategies will be used to translate light-dependent capture of the adapter peptide into receptor activation or inhibition. These include dimerizing the adapter peptide with a dimeric photoswitch to activate growth factor receptors, and delivering the adapter peptide, including the ligand, to a G-protein coupled receptor via an antibody-tethered photoswitch. Other strategies may be developed to activate different types of receptors and ion channels. We will test the effectiveness of the Universal Photoswitch approach on two example receptors: the TrkB receptor for brain-derived growth factor (BDNF), a member of the neurotrophin family of receptor tyrosine kinases, and the receptor for neuropeptide Y, which is a GPCR-type receptor. Generating a method for light-sensitive regulation of these receptors will allow examination of their roles in development and neural function in intact tissue with unprecedented precision, but more importantly, it will demonstrate the emergence of a powerful new technique for receptor regulation that can be applied to any cell surface protein for which a known peptide ligand exists. PUBLIC HEALTH RELEVANCE: Our goal is to develop a single photochemical switch that can be interfaced in a combinatorial manner with many types of cell surface receptors to enable precise temporal, spatial, and biochemical control over neuronal functions. The NIDA/NIMH/NINDS EUREKA applications were reviewed differently from more traditional NIH grant mechanisms. Specifically, the review process consisted of two phases. During the first (i.e., electronic) phase a selected panel of reviewers were given the following guidelines by which to assess the applications. They were asked to determine whether they: Strongly Agree, Moderately Agree, Neither Agree nor Disagree, Moderately Disagree, or Strongly Disagree with these descriptions. Their ratings and any additional comments are below. These initial ratings also provided the basis for the review panel to determine whether an application would be discussed during an in person meeting. Because of the very stringent review criteria and limited pool of funds set aside for this program, the review panel chose only to discuss applications that garnered the most enthusiasm. The Resume and Summary of the Discussion above summarizes opinions of the person meeting and forms the basis of the final score. Significance: This study addresses an important problem and the outcome of the proposed studies will drive the field. The potential impact of the proposed research is exceptional, in terms of the magnitude of the impact and the size of the community affected. Innovation: The project is highly original and exceptionally innovative and seriously challenges existing paradigms or clinical practice. The project addresses a major barrier to progress in the field or it develops or employs exceptionally novel concepts, approaches, methodologies, tools, or technologies. Approach: The logic of the approach is sufficiently compelling despite the lack of experimental detail. The conceptual (or clinical) framework, design, methods, and analyses are adequately developed, well integrated and reasoned, and are appropriate for the aims of the project. The applicant acknowledges potential problem areas and considers alternative tactics. The information in the timeline inspires confidence that the PI will be able to document progress in each year of the award and either complete the project or demonstrate conclusively that it cannot be completed, despite good-faith efforts, during the term of the award. The requested duration of the award is appropriate for the proposed research. Investigators: The PD/PI(s) and other key personnel are appropriately trained and well-suited to carry out this work. Past achievements of the PI(s) suggest that the investigator(s) is/are exceptionally innovative and likely to make paradigm-shifting, high-impact discoveries. If the PI does not have a history of doing exceptionally innovative, high-impact research, the logic of the experimental plan suggests that there is at least some likelihood of success. The project is high priority for the PI(s), as indicated by the person-months of effort that the PI(s) will devote to it. For applications designating multiple PDs/PIs, the leadership plan, including the designated roles and responsibilities, governance, and organizational structure, are consistent with and justified by the aims of the project and the expertise of each of the PDs/PIs. Environment: The scientific environment(s), in which the work will be performed, contributes to the probability of success. The proposed studies benefit from unique features of the scientific environment, subject populations, or employ useful collaborative arrangements. There is evidence of institutional support.

描述（由申请人提供）： 近年来，生物学上最令人兴奋的技术发展之一是光化学方法的出现，用于控制光线活性。我们在该项目中的目标是通过生成在广泛的细胞表面受体和离子通道上赋予光灵敏度的工具来开发一种可控制细胞功能其他方面的广泛适用方法。为了应对这一挑战，我们将使用模块化方法，利用单个“通用Photoswitch”作为关键的光感应组件。 Photoswitch在其核心中包含了小的异构偶氮苯一部分，该部分分别缩短并延长了380和500 nm光的响应。我们将使用此照片开关通过“衔接肽”间接调节受体和通道活动，该肽包含一个“捕获结构域”，该肽识别photoswitch的短构型，但不识别Photoswitch的长配置，以及一个包含靶标细胞表面受体或ion ion ion Channel的肽激活剂或抑制剂的配体域。在所有衔接肽中，捕获结构域保持恒定，从而可以通过单个通用照片开关进行控制，但是配体域的性质是量身定制的，以调节特定的受体。将使用几种策略将衔接肽的光依赖捕获转化为受体激活或抑制作用。这些包括用二聚体的照片开关将衔接肽二聚二聚体以激活生长因子受体，并通过抗体螺旋螺旋射线开关将衔接肽（包括配体）传递到G蛋白偶联受体中。可以开发其他策略来激活不同类型的受体和离子通道。我们将在两个示例受体上测试通用照片开关方法的有效性：脑衍生生长因子的TRKB受体（BDNF），受体酪氨酸激酶的神经营养蛋白家族的成员和神经肽Y的受体，是GPCR型受体。生成一种对这些受体的光敏调节的方法，将允许以前所未有的精度来检查其在完整组织中的发育和神经功能中的作用，但更重要的是，它将证明可以将有力的受体调节的新技术出现在任何可应用于任何已知肽座的细胞表面蛋白质上。 公共卫生相关性： 我们的目标是开发单个光化学开关，该开关可以与许多类型的细胞表面受体相连，以实现对神经元功能的精确时间，空间和生化控制。 NIDA/NIMH/NINDS EUREKA应用程序的审查与更传统的不同 NIH赠款机制。具体而言，审查过程由两个阶段组成。在 首先，（即电子）阶段A选定的审阅者面板由以下指南。 评估申请。他们被要求确定他们是否：完全同意， 中度同意，既不同意也不同意，中度不同意或强烈不同意 这些描述。他们的评分和任何其他评论如下。这些初始评分 还为审核面板提供了确定申请是否将是的基础 在一次会议上讨论。由于非常严格的评论标准和有限 为该计划拨出的资金库，评论面板只选择讨论申请 这赢得了最热情。上面讨论的简历和摘要 总结人会议的意见，并构成最终得分的基础。 意义：本研究解决了一个重要的问题，拟议研究的结果将 驱动场。就幅度而言，拟议研究的潜在影响是非凡的 社区影响的影响和大小受到影响。 创新：该项目是高度原始和异常创新和严重挑战的现有 范式或临床实践。该项目解决了该领域进步或IT的主要障碍 开发或采用异常新颖的概念，方法，方法，工具或技术。 方法：尽管缺乏实验细节，但该方法的逻辑足以令人信服。 概念（或临床）框架，设计，方法和分析已充分发展，很好 集成和推理，适合项目的目的。申请人承认 潜在的问题领域并考虑替代策略。时间表中的信息灵感 对PI的信心将能够记录奖励的每一年的进度并完成 该项目或最终证明，尽管有良好的努力，但在 奖项的期限。该奖项的持续时间适合拟议的研究。 调查人员：PD/PI（S）和其他关键人员经过适当培训，适合携带 出去这项工作。 PI的过去成就表明，研究者是/异常的 创新的，可能会带来范式转移，高影响力的发现。如果PI没有 进行异常创新，高影响研究的历史，实验计划的逻辑 表明至少有成功的可能性。该项目是PI的优先事项 PI将致力于这一努力的努力表明。用于指定的应用 多个PDS/PI，领导计划，包括指定的角色和职责，治理， 和组织结构与项目的目的一致和合理 每个PDS/PI的专业知识。 环境：将执行工作的科学环境，有助于 成功的概率。拟议的研究受益于科学环境的独特特征， 主题人群，或采用有用的协作安排。有制度的证据 支持。