Exploring the role of HINT1 protein in neuronal function

描述(申请人提供)：蛋白激酶C相互作用蛋白(PKCI/HINT1)是一种属于组氨酸三联体(HIT)家族的小蛋白。尸检研究已确认PKCI/HINT1是精神分裂症和双相情感障碍的候选分子。在我们实验室的研究中，PKCI/HINT1 KO小鼠表现出一种行为和内分泌特征的表型，表明情绪功能的变化，包括抗焦虑和抗抑郁样行为以及对苯丙胺的超敏。这些结果提示，PKCI/HINT1在正常的中枢神经系统功能和精神疾病状态中起着关键作用。目前的R21方案旨在为在细胞水平上表征PKCI/HINT1功能的长期研究奠定基础，并开发该蛋白的新抑制剂以便于表征，并有可能开发成治疗相关中枢神经系统疾病的新型治疗药物。具体目标包括：目的1.建立PKCI/HINT1的细胞分析方法，重点研究与神经元存活、神经元生长和钙信号相关的潜在活性。化验开发将强调高通量能力，以促进未来的药物发现和优化工作。目的2.利用基于结构的计算药物设计方法，寻找与PKCI/HINT1结合几率较高的低分子化合物。然后，将使用体外荧光和ITC分析，并辅之以核磁共振分析，来分析已鉴定的化合物与PKCI/HINT1结合的能力。从本质上讲，这项工作将填补已知的PKCI/HINT1三维结构与其已知的体内行为特征之间的空白。在拟议的研究中获得的关于PKCI/HINT1细胞功能的知识将填补这一空白，从而允许未来对PKCI/HINT1的药理学进行广泛的研究，可能导致开发出一种合理的药物设计方法来开发治疗情绪障碍的新型药物。 公共卫生相关性：这项研究旨在通过两组实验了解一种新的蛋白质(HINT1)在大脑中的功能。一个是研究HINT1对神经元型细胞的生长和死亡有什么影响，另一个是寻找可能与HINT1相互作用的小化学物质。

DESCRIPTION (provided by applicant): Protein kinase C interacting protein (PKCI/HINT1) is a small protein belonging to the histidine triad (HIT) family of proteins. Postmortem studies have identified PKCI/HINT1 as a candidate molecule for schizophrenia and bipolar disorder. In studies from our laboratory, PKCI/HINT1 KO mice displayed a phenotype of behavioral and endocrine features which indicate changes of mood function, including anxiolytic- like and anti-depressant like behaviors and supersensitivity to amphetamine. These results suggest that PKCI/HINT1 plays a key role in normal CNS function and state of mental diseases. The present R21 proposal is designed to set the groundwork for long-term studies characterizing the function of PKCI/HINT1 at the cellular level and developing novel inhibitors of the protein to facilitate the characterization as well as have the potential to be developed into novel therapeutic agents for related CNS disorders. The specific aims include: Aim 1. Develop cellular assays for PKCI/HINT1 focusing on potential activities associated with neuronal survival, neuronal growth and calcium signaling. Assay development will emphasize high- throughput capabilities to facilitate future drug discovery and optimization efforts. Aim 2. Identify low molecular weight compounds with a high probability of binding to PKCI/HINT1 using structure-based computational drug design methods. Identified compounds will then be assayed for their ability to bind PKCI/HINT1 using in vitro fluorescence and ITC assays, supplemented with NMR analysis. In essence, the proposed work will fill a gap between the known 3D structure of PKCI/HINT1 and its known in vivo behavioral characteristics. Knowledge of the cellular function of PKCI/HINT1 gained in the proposed study will fill this gap allowing for extensive future studies of the pharmacology of PKCI/HINT1, potentially leading to a rational drug design approach to the development of novel therapeutic agents for mood disorders. PUBLIC HEALTH RELEVANCE: The study is to understand a novel protein (HINT1) function in brain through two sets of experiments. One is to examine what HINT1 do on growth and death of neuronal type of cells and the other is to look for small chemicals that may interact with HINT1.