Analysis of circadian profiles in cultured fibroblasts

描述(申请人提供)：复发性情感疾病的一些最突出和最典型的症状，例如双相和单相障碍，包括睡眠中的戏剧性干扰：觉醒或休息：活动周期。已经证明，控制休息：活动周期可以深刻地改变这些疾病的临床表现的过程。这些发现表明，睡眠：清醒或休息变化的遗传易感性：活动周期代表了复发性情感障碍的内在表型。根据最近的发现，基因组中5%-10%的基因表达受Clock控制(Akhtar等人，2002；Panda等人，2002)，我们认为Clock基因和/或由Clock控制的基因可能是双相情感障碍的多基因遗传基础。最近发现，培养的皮肤成纤维细胞在血清休克(50%胎牛血清)后表现出核心时钟基因的同步循环表达(Balsalobre等人，1998；Nagoshi等人，2004)为研究双相情感障碍和昼夜节律系统之间的联系提供了新的途径。这项应用的目的是探索使用双相情感障碍患者皮肤活检组织中的成纤维细胞来研究其内在生物钟的可能性。具体来说，我们的目标是：1.建立一种高通量监测培养的人成纤维细胞昼夜节律的方法，通过实时定量聚合酶链式反应和生物发光记录来检测核心时钟和时钟控制基因的循环表达；以及目的2.应用该方法来检测核心时钟机制是否参与双相情感障碍的病理生理。在这个目标中，将包括通过Coriell细胞库获得的成纤维细胞细胞系的分析(重要注意：作为原始细胞来源的特定个体的身份对于该库来说是未知的)。这一高度探索性的项目具有巨大的潜力，可以加速我们对一种重要精神障碍的遗传基础的理解。

DESCRIPTION (provided by applicant): Some of the most prominent and characteristic symptoms of recurrent affective illnesses, e.g., bipolar and unipolar disorders, include dramatic disturbances in the sleep:wake or rest:activity cycle. It has been shown that manipulations of rest:activity cycles can profoundly change the course of clinical manifestations of these illnesses. These findings suggest that genetic predisposition to alterations in sleep:wake or rest:activity cycles represent endophenotypes for recurrent affective disorders. Based on recent findings that gene expression of 5-10% of genes in the genome are controlled by the clock (Akhtar et al., 2002; Panda et al., 2002), we propose that clock genes and/or genes that are controlled by the clock may underlie polygenic inheritance of the bipolar disorder. Recent findings that cultured skin fibroblasts, following a serum shock (50% fetal bovine serum) exhibit synchronized cycling in the expression of core clock genes (Balsalobre et al., 1998; Nagoshi et al., 2004) provide a new avenue to investigate the link between bipolar illness and the circadian system. The purpose of this application is to explore the possibility of using fibroblasts derived from skin biopsies of bipolar patients to investigate their intrinsic circadian clock. Specifically, our aims are: Aim 1. To develop a high-throughput method for monitoring circadian rhythms in human cultured fibroblasts to examine cycling expression of core clock and clock-controlled genes by Real-time quantitative PCR and bioluminescence recording; and Aim 2. To apply the method to examine if the core clock machinery is involved in the pathophysiology of bipolar disorder. In this aim will involve analysis of fibroblasts cell-lines available through the Coriell Cell Repositories (Important note: The identity of the specific individuals that were the source of the original cells is not known to the repository).This highly exploratory project has a tremendous potential for accelerating our understanding of the genetic basis of an important mental disorder.