Analysis of circadian profiles in cultured fibroblasts

描述（由申请人提供）：一些复发性情感疾病的最突出和最典型的症状，例如双相情感障碍和单相情感障碍，包括睡眠：清醒或休息：活动周期中的剧烈干扰。研究表明，对休息活动周期的操纵可以深刻地改变这些疾病的临床表现。这些发现表明，遗传易感性的睡眠：觉醒或休息：活动周期的改变代表了复发性情感障碍的内表型。根据最近的研究发现，基因组中5-10%的基因表达受生物钟控制（Akhtar et al., 2002; Panda et al., 2002），我们提出生物钟基因和/或受生物钟控制的基因可能是双相情感障碍多基因遗传的基础。最近的研究发现，在血清休克（50%胎牛血清）后，培养的皮肤成纤维细胞在核心时钟基因的表达中表现出同步循环（Balsalobre等人，1998年；Nagoshi等人，2004年），为研究双相情感障碍与昼夜节律系统之间的联系提供了新的途径。本应用的目的是探索利用从双相患者皮肤活检中提取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：利用实时荧光定量PCR和生物发光记录技术，建立一种高通量监测人培养成纤维细胞昼夜节律的方法，检测核心时钟和时钟控制基因的循环表达；和Aim 2。应用该方法研究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，将涉及分析通过科里尔细胞库提供的成纤维细胞细胞系（重要注意：库不知道原始细胞来源的特定个体的身份）。这个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疾病的遗传基础的理解。

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.