Analysis of circadian profiles in cultured fibroblasts

描述（由申请人提供）：复发性情感疾病的一些最突出和特征性症状，例如，双相和单极障碍，包括睡眠：清醒或休息：活动周期中的剧烈干扰。它已被证明，操作休息：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，遗传易感性改变睡眠：觉醒或休息：活动周期代表内表型复发性情感障碍。基于基因组中5-10%基因的基因表达受时钟控制的最近发现（Akhtar et al.，2002; Panda等人，2002），我们提出时钟基因和/或由时钟控制的基因可能是双相情感障碍的多基因遗传的基础。最近的发现是，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表达的同步循环（Balsalobre等人，1998; Nagoshi等人，2004）提供了一个新的途径来调查双相情感疾病和昼夜节律系统之间的联系。本申请的目的是探索使用双相情感障碍患者皮肤活检来源的成纤维细胞来研究其内在生物钟的可能性。具体而言，我们的目标是：目标1。开发一种高通量方法，用于监测培养的人成纤维细胞中的昼夜节律，以通过实时定量PCR和生物发光记录来检查核心时钟和时钟控制基因的循环表达;以及目的2。应用该方法检测核心时钟机制是否参与双相情感障碍的病理生理过程。在这一目标将涉及到通过Coriell细胞库可用的成纤维细胞系的分析（重要说明：作为原始细胞来源的特定个体的身份不为库所知）。这个高度探索性的项目具有巨大的潜力，可以加速我们对重要精神障碍遗传基础的理解。

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.