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）： 近年来生物学领域最令人兴奋的技术发展之一是用光控制电活动的光化学方法的出现。我们在这个项目中的目标是通过生成赋予广泛的细胞表面受体和离子通道光敏感性的工具来开发一种广泛适用的方法来控制细胞功能的其他方面。为了应对这一挑战，我们将采用模块化方法，利用单个“通用光电开关”作为关键的光传感组件。该光电开关的核心包含小的可异构化偶氮苯部分，它分别响应 380 和 500 nm 的光而缩短和延长。我们将使用这种光开关来间接调节受体和通道活性，通过“适配器肽”，它包含一个“捕获结构域”，它识别光开关的短结构，但不识别长结构，以及一个配体结构域，它包含目标细胞表面受体或离子通道的肽激活剂或抑制剂。所有衔接肽中的捕获结构域保持恒定，允许通过单个通用光开关进行控制，但配体结构域的性质是为了调节特定受体而定制的。将使用几种策略将接头肽的光依赖性捕获转化为受体激活或抑制。这些包括用二聚体光开关将衔接肽二聚化以激活生长因子受体，以及通过抗体系连的光开关将包括配体在内的衔接肽递送至G蛋白偶联受体。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光电开关方法的有效性：脑源性生长因子 (BDNF) 的 TrkB 受体（受体酪氨酸激酶神经营养蛋白家族的成员）和神经肽 Y 的受体（GPCR 型受体）。产生一种对这些受体进行光敏调节的方法将能够以前所未有的精度检查它们在完整组织中的发育和神经功能中的作用，但更重要的是，它将证明一种强大的受体调节新技术的出现，该技术可应用于存在已知肽配体的任何细胞表面蛋白。 公共卫生相关性： 我们的目标是开发一种单一的光化学开关，它可以以组合方式与多种类型的细胞表面受体连接，从而实现对神经元功能的精确时间、空间和生化控制。 NIDA/NIMH/NINDS EUREKA 申请的审核方式与传统申请不同 NIH 资助机制。具体来说，审查过程分为两个阶段。期间 第一（即电子）阶段，选定的评审小组获得了以下指导方针： 用于评估应用程序。他们被要求确定是否： 强烈同意， 比较同意、既不同意也不反对、比较不同意或强烈不同意 这些描述。他们的评级和任何其他评论如下。这些初始评级 还为审查小组确定申请是否得到批准提供了依据 在面对面会议期间进行讨论。由于审查标准非常严格且数量有限 为该计划预留的资金池，审查小组选择仅讨论申请 这引起了最大的热情。上述讨论的简历和总结 总结会议成员的意见，形成最终评分的基础。 意义：这项研究解决了一个重要问题，拟议研究的结果将 驱动领域。就规模而言，拟议研究的潜在影响是非凡的 影响以及受影响社区的规模。 创新：该项目具有高度原创性和非凡的创新性，严重挑战现有的 范式或临床实践。该项目解决了该领域取得进展的主要障碍 开发或采用异常新颖的概念、途径、方法、工具或技术。 方法：尽管缺乏实验细节，但该方法的逻辑足够引人注目。 概念（或临床）框架、设计、方法和分析得到充分发展、良好 综合且合理，并且适合项目的目标。申请人承认 潜在的问题领域并考虑替代策略。时间轴上的信息鼓舞人心 相信 PI 将能够记录获奖每一年的进展，并且完成 该项目或最终证明，尽管有诚意的努力，该项目仍无法完成 奖励期限。所要求的奖励期限适合拟议的研究。 调查人员：PD/PI 和其他关键人员经过适当的培训并且非常适合携带 出这个作品。 PI 过去的成就表明研究者非常出色 创新并可能做出范式转变、高影响力的发现。如果 PI 没有 进行异常创新、高影响力研究的历史、实验计划的逻辑 表明至少有一定的成功可能性。该项目对于 PI 来说具有高度优先级，因为 由PI（们）为此付出的人月努力来表示。对于指定的应用 多个 PD/PI、领导力计划，包括指定的角色和职责、治理、 和组织结构，与项目和项目的目标一致并合理 每个 PD/PI 的专业知识。 环境：开展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， 受试者群体，或采用有用的合作安排。有证据表明制度存在 支持。